Alternative proteins for meat and dairy replacers: Food safety and future trends.

Traditionally, meat and dairy products have been important protein sources in the human diet. Consumers are eating more plant-based proteins, which is reflected in current market trends. Assessing how alternative proteins are processed and their impact on food safety helps realize market opportunities while ensuring food safety. In this review, an analysis of the food safety hazards, along with current industry trends and processing methods associated with alternative proteins for meat and dairy products for the European Union market is described. Understanding the effects of processing and safety alternative proteins is paramount to ensuring food safety and understanding the risks to consumers. However, the data here is limited. With the expected further increase in protein alternatives in consumers' diets, the risk of food allergens is apparent. The occurrence of processing contaminants in plant-based alternatives may occur, along with anti-nutritional compounds, which interfere with the absorption of nutrients. Further, typical food safety hazards related to the plant, the product itself, or processing are relevant. Although hazards in insects and seaweed are being addressed, other protein alternatives like cultured meat and SCPs warrant attention. Our findings can aid industry and governmental authorities in understanding current trends and prioritizing hazards for future monitoring.

Food safety hazards in the European seaweed chain.

Seaweed is a source of protein that can help overcome the anticipated challenges of a growing world population and the current challenges for finding alternatives for animal proteins in the Western diet. Thus far, data on the safety of seaweed for feed and food purposes in the Western world are scattered. This study aimed to review the available knowledge on the presence of food safety hazards in seaweed, including factors influencing their presence, and to prioritize the hazards that may pose a risk to human health. Given current knowledge from the literature, data from the Rapid Alert System for Food and Feed, and results from a stakeholder survey, 22 food safety hazards were ranked into major (4), moderate (5), and minor (13) hazards. Arsenic, cadmium, iodine, and Salmonella were identified as major hazards. Hazards, where data gaps exist, should be carefully assessed. These include pesticide residues, dioxins and polychlorinated biphenyls, brominated flame retardants, polycyclic aromatic hydrocarbons, pharmaceuticals, marine biotoxins, allergens, micro- and nanoplastics, other pathogenic bacteria, norovirus, and hepatitis E virus. It is recommended to collect more data on these hazards in future studies. Many factors can affect the presence of hazards including seaweed type, physiology, season, harvest and cultivation environment, geography including the location of cultivation, alongside further processing. Moreover, when seaweed is cultivated near industrialized or anthropogenic activities, these activities may negatively influence water quality, which can increase the likelihood of hazards in seaweed. Results of the ranking prioritized hazards can be used to prioritize monitoring programs and adjusted given future additional knowledge covering the data gaps.

Microbiological Reduction Strategies of Irrigation Water for Fresh Produce.

ABSTRACT: Irrigation water can be a source of pathogenic contamination of fresh produce. Controlling the quality of the water used during primary production is important to ensure food safety and protect human health. Several measures to control the microbiological quality of irrigation water are available for growers, including preventative and mitigation strategies. However, clear guidance for growers on which strategies could be used to reduce microbiological contamination is needed. This study evaluates pathogenic microorganisms of concern in fresh produce and water, the microbiological criteria of water intended for agricultural purposes, and the preventative and mitigative microbial reduction strategies. This article provides suggestions for control measures that growers can take during primary production to reduce foodborne pathogenic contamination coming from irrigation water. Results show that controlling the water source, regime, and timing of irrigation may help to reduce the potential exposure of fresh produce to contamination. Moreover, mitigation strategies like electrolysis, ozone, UV, and photocatalysts hold promise either as a single treatment, with pretreatments that remove suspended material, or as combined treatments with another chemical or physical treatment(s). Based on the literature data, a decision tree was developed for growers, which describes preventative and mitigation strategies for irrigation-water disinfection based on the fecal coliform load of the irrigation water and the water turbidity. It helps guide growers when trying to evaluate possible control measures given the quality of the irrigation water available. Overall, the strategies available to control irrigation water used for fresh produce should be evaluated on a case-by-case basis because one strategy or technology does not apply to all scenarios.

Effectiveness of a peracetic acid solution on Escherichia coli reduction during fresh-cut lettuce processing at the laboratory and industrial scales.

Fresh leafy greens like lettuce can be consumed raw and are susceptible to foodborne pathogens if they become contaminated. Recently, the number of reported pathogenic foodborne outbreaks related to leafy greens has increased. Therefore, it is important to try to alleviate the human health burden associated with these outbreaks. Processing of fresh-cut lettuce, including washing, is a step in the supply chain that needs to be well controlled to avoid cross-contamination. Current measures to control the quality of lettuce during washing include the use of chemicals like chlorine; however, questions regarding the safety of chlorine have prompted research for alternative solutions with peracetic acid (PAA). This study evaluates the effectiveness of a PAA (c.a. 75 mg/L) solution on the reduction of a commensal E. coli strain during the washing of fresh-cut lettuce. Experiments were performed at the laboratory scale and validated at the industrial scale. We observed that the use of PAA was not adversely affected by the organic load in the water. The contact time and dose of the PAA showed to be relevant factors, as observed by the approximately 5-log reduction of E. coli in the water. Results showed that once introduced during washing, E. coli remained attached to the lettuce, thus supporting the need to control for pathogenic bacteria earlier in the supply chain (e.g., during primary production) as well as during washing. Moreover, our results showed that the use of PAA during washing did not have an apparent effect on the levels of fluorescent pseudomonads (FP) and total heterotrophic bacteria (THB) in lettuce. Overall, our results at the laboratory and industrial scales confirmed that during the processing of fresh-cut produce, where the accumulation of soil, debris, and other plant exudates can negatively affect washing, the use of a PAA (c.a. 75 mg/L) solution was an effective and safe wash water disinfectant that can potentially be used at the industrial scale.

An analysis of the influence of various tenderising treatments on the tenderness of meat from Polish Holstein-Friesian bulls and the course of changes in collagen.

The aim of the study was to analyse the influence of tenderising treatments applied to the carcasses of Polish Holstein-Friesian (PHF) bulls of Black-and-White variety on the process of meat tenderisation and to assess the role of collagen in this process. The research was carried out on m. longissimus thoracis et lumborum. The carcasses were subjected to high-voltage electrical stimulation (ES), conditioning (CD), and both treatments together (ES + CD). The carcasses which were only refrigerated were the control group. The content of collagen in meat, its solubility, the share of the polypeptide subunits α1(I)CB7 and α1(I)CB8 of type I collagen and α1(III)CB5 of type III collagen were also analysed. ES with and without CD significantly accelerated the meat tenderisation and increased collagen solubility. CD always caused the degradation of type I collagen subunits, especially the α1(I)CB7 subunit. However, CD had significantly lesser influence on the rate of meat tenderisation than ES.

High-dose intravenous methylprednisolone therapy in patients with Graves' orbitopathy is associated with the increased activity of factor VIII.

PURPOSE: Venous thromboembolic events (VTE), with their life-threatening manifestation as pulmonary embolism, are potential adverse effects of intravenous methylprednisolone (IVMP) pulse therapy, partially due to a hypercoagulable state. The aim of the study was to analyze the influence of IVMP on selected hemostatic parameters in patients with moderate-to-severe Graves' orbitopathy (GO). METHODS: 26 euthyroid patients with GO were treated with 12 pulses of IVMP (6 × 0.5, 6 × 0.25 g every week). Hemostatic variables [factor (F) II, FV, FVII, FVIII, fibrinogen, antithrombin, activated partial thromboplastin time (aPTT), prothrombin time, international normalized ratio of prothrombin time, platelets and D-dimer] were analysed before, 24 and 48 h after 1st, 6th and 12th pulse. RESULTS: A constant, transient trend in changes of some hemostatic variables was observed after all assessed pulses. We discovered an increase in median activity of FVIII 24 and 48 h after pulses, with a shortening of aPTT 24 h after each IVMP pulse (p < 0.00005). FVII decreased 24 h after each pulse (p < 0.0005 after 1st and 12th, p < 0.00005 after 6th). Fibrinogen level decreased 48 h after each pulse (P < 0.00005). We did not observe any statistically significant changes in hemostatic parameters in the long-term evaluation. Therapy was concluded in one patient after the 9th pulse due to pulmonary embolism. CONCLUSIONS: The increase of FVIII activity is a consequence of treatment with IVMP and occurs after each pulse. In patients with additional risk factors of VTE, anticoagulation prophylaxis should be considered.

Procalcitonin (PCT) Predicts Worse Outcome in Patients with Chronic Heart Failure with Reduced Ejection Fraction (HFrEF).

INTRODUCTION: Procalcitonin (PCT) is an excellent marker of sepsis but was not extensively studied in cardiology. The present study investigated PCT plasma concentration in patients with chronic heart failure with reduced ejection fraction (HFrEF) and its prognostic value during 24-month follow-up. MATERIAL AND METHODS: Study group consisted of 130 patients with HFrEF (LVEF ≤ 45%) and 32 controls. PCT level was assessed on admission in all patients. Telephone follow-up was performed every three months over a period of 2 years. Endpoints were death of all causes and readmission for HFrEF exacerbation. RESULTS: HFrEF patients had significantly higher PCT concentration than controls (166.95 versus 22.15 pg/ml; p < 0.001). Individuals with peripheral oedema had increased PCT comparing to those without oedema (217.07 versus 152.12 pg/ml; p < 0.02). In ROC analysis, PCT turned out to be a valuable diagnostic marker of HFrEF (AUC 0.91; p < 0.001). Kaplan-Meier survival curves revealed that patients with PCT in the 4th quartile had significantly lower probability of survival than those with PCT in the 1st and 2nd quartiles. In univariate, but not multivariate, analysis, procalcitonin turned out to be a significant predictor of death during 24-month follow-up. (HR 1.002; 95% CI 1.000-1.003; p < 0.03). CONCLUSIONS: Elevated PCT concentration may serve as another predictor of worse outcome in patients with HFrEF.

Efficacy of chlorine dioxide on Escherichia coli inactivation during pilot-scale fresh-cut lettuce processing.

Controlling water quality is critical in preventing cross-contamination during fresh produce washing. Process wash water (PWW) quality can be controlled by implementing chemical disinfection strategies. The aim of this study was to evaluate the pilot-scale efficacy of chlorine dioxide (ClO2) during processing on the reduction of Escherichia coli in the PWW and on processed fresh-cut 'Lollo Rossa' lettuce. The objective was to have a residual target concentration of either 5 or 3 mg/L ClO2 in the washing tank (3.5 m3) before and during 800 kg of lettuce processing (90 min). After 90 min., a nonpathogenic, non-Extended Spectrum Beta-Lactamase (ESBL) E. coli inoculum from an overnight culture broth (37 °C) was added to the tank resulting in an approximate final level of 106 CFU/mL. PWW and lettuce samples for microbiological and chemical analyses were taken before and after the input and supply halted. ClO2 concentrations quickly decreased after ClO2 input halted, yet a residual concentration of ≥2.5 mg/L and ≥2.1 mg/L ClO2, respectively for 5 and 3 mg/L pilots, was present 12 min after the supply halted. No detectable levels of E. coli (limit of detection 5 log) were determined in the water within 1 min after E. coli was added to the ClO2 containing wash water. Results demonstrated that ClO2 use at the semi-commercial pilot scale was able to reduce the E. coli peak contamination in the PWW. After storage (5 days, 4 °C), background microbial communities (i.e., fluorescent Pseudomonads and total heterotrophic bacteria) grew out on lettuce. Overall, ClO2 decreased the potential for cross-contamination between batches compared to when no sanitizer was used. Chlorate levels of the lettuce sampled before entering the wash water ranged from 7.3-11.6 µg/kg. The chlorate levels of the lettuce sampled after being washed in the ClO2 containing wash water, as well as after rinsing and centrifugation, ranged from 22.8-60.4 µg/kg; chlorite levels ranged from 1.3-1.6 mg/kg, while perchlorate levels were below the limit of quantification (LOQ, <5 ng/g). In this study, we report the semi-commercial pilot-scale evaluation of ClO2, for its ability to maintain the PWW quality and to prevent cross-contamination in the washing tank during fresh-cut lettuce processing. Furthermore, we provide quantitative values of ClO2 disinfection by-products chlorate and chlorite as well as of perchlorate from PWW and/or lettuce samples.

Particle Size of Milk Protein Concentrate Powder Affects the Texture of High-Protein Nutrition Bars During Storage.

Milk protein concentrate powder with 85% protein (MPC85) was jet-milled to give 2 particle size distributions (that is, JM-Coarse and JM-Fine) or freeze-dried (FD), in order to improve the functional properties of MPC85 for use in high-protein nutrition (HPN) bars. Volume-weighted mean diameter decreased from 86 µm to 49, 22, and 8 µm in FD, JM-Coarse, and JM-Fine, respectively (P < 0.05). The MPC85 powders modified by jet-milling and freeze-drying were significantly denser than the control MPC85 (P < 0.05). Volume of occluded air in the modified powders decreased (P < 0.05) by an order of magnitude, yet only FD possessed a lower volume of interstitial air (P < 0.05). Particle size reduction and freeze-drying MPC85 decreased its water holding capacity and improved its dispersibility by at least 20%. Contact angle measurements showed that these modifications increased initial hydrophobicity and did not improve wettability. HPN bars made from JM-Fine or FD were firmer by 40 or 17 N, respectively, than the control on day 0 (P < 0.05). HPN bar maximum compressive force increased by 38%, 33%, and 242% after 42 d at 32 °C when formulated with JM-Fine, FD, or control MPC85, respectively. HPN bars prepared with JM-Fine were less crumbly than those formulated with control or FD MPC85. Physically altering the particle structure of MPC85 improved its ability to plasticize within HPN bars and this improved their cohesiveness and textural stability.

Textural performance of crosslinked or reduced-calcium milk protein ingredients in model high-protein nutrition bars.

Transglutaminase (Tgase) crosslinking and calcium reduction were investigated as ways to improve the texture and storage stability of high-protein nutrition (HPN) bars formulated with milk protein concentrate (MPC) and micellar casein concentrate (MCC). The MPC and MCC crosslinked at none, low, and high levels, and a reduced-calcium MPC (RCMPC) were each formulated into model HPN bars. Hardness, crumbliness, moisture content, pH, color, and water activity of the HPN bars were measured during accelerated storage. The HPN bars prepared with MPC were harder and more cohesive than those prepared with MCC. Higher levels of Tgase crosslinking improved HPN bar cohesiveness and decreased hardening during storage. The RCMPC produced softer, yet crumblier HPN bars. Small textural differences were observed for the HPN bars formulated with the transglutaminase crosslinked proteins or RCMPC when compared with their respective controls. However, modification only slightly improved protein ingredient ability to slow hardening while balancing cohesion and likely requires further improvement for increased applicability in soft-texture HPN bars.

Instrumental and Sensory Texture Attributes of High-Protein Nutrition Bars Formulated with Extruded Milk Protein Concentrate.

Previous instrumental study of high-protein nutrition (HPN) bars formulated with extruded milk protein concentrate (MPC) indicated slower hardening compared to bars formulated with unmodified MPC. However, hardness, and its change during storage, insufficiently characterizes HPN bar texture. In this study, MPC80 was extruded at 2 different conditions and model HPN bars were prepared. A trained sensory panel and instrumental techniques were used to measure HPN bar firmness, crumbliness, fracturability, hardness, cohesiveness, and other attributes to characterize texture change during storage. Extrusion modification, storage temperature, and storage time significantly affected the instrumental and sensory panel measured texture attributes. The HPN bars became firmer and less cohesive during storage. When evaluated at the same storage conditions, the texture attributes of the HPN bars formulated with the different extrudates did not differ significantly from each other. However, textural differences were noted most of the time between the control and the HPN bars formulated with extruded MPC80. An adapted HPN bar crumbliness measurement technique produced results that were correlated with sensory panel measured crumbliness (r = 0.85) and cohesiveness (r = -0.84). Overall, the HPN bars formulated with extruded MPC80 were significantly softer, less crumbly, and more cohesive than the control during storage.

Microstructural Changes in High-Protein Nutrition Bars Formulated with Extruded or Toasted Milk Protein Concentrate.

Milk protein concentrates with more than 80% protein (that is, MPC80) are underutilized as the primary protein source in high-protein nutrition bars as they impart crumbliness and cause hardening during storage. High-protein nutrition bar texture changes are often associated with internal protein aggregations and macronutrient phase separation. These changes were investigated in model high-protein nutrition bars formulated with MPC80 and physically modified MPC80s. High-protein nutrition bars formulated with extruded MPC80s hardened slower than those formulated with toasted or unmodified MPC80. Extruded MPC80 had reduced free sulfhydryl group exposure, whereas measurable increases were seen in the toasted MPC80. High-protein nutrition bar textural performance may be related to the number of exposed free sulfhydryl groups in MPC80. Protein aggregations resulting from ingredient modification and high-protein nutrition bar storage were studied with sodium dodecyl sulfate polyacrylamide gel electrophoresis. Disulfide-based protein aggregations and changes in free sulfhydryl concentration were not consistently relatable to high-protein nutrition bar texture change. However, the high-protein nutrition bars formulated with extruded MPC80 were less prone to phase separations, as depicted by confocal laser scanning microscopy, and underwent less texture change during storage than those formulated with toasted or unmodified MPC80.

Characterization of extruded and toasted milk protein concentrates.

Important functional properties of milk protein concentrate with 80% protein (MPC80), modified with low- and high-shear extrusion, or low-temperature toasting were compared. The effect of high- and low-shear profile screws in a corotating twin-screw extruder, and 4 different ramped temperature profiles with die temperatures of 65, 75, 90, and 120 °C were compared. Extrudates were pelletized, dried, and ground to a fine powder. Toasting was done at 75 and 110 °C for 4 h for milk protein modification. Extruded and toasted MPC80 had reduced protein solubility and surface hydrophobicity. Extrusion decreased water-holding capacity (WHC). Toasted MPC80 had increased WHC when treated at 75 °C, but WHC decreased when heated at 110 °C. The treatments had no strong influence on gel strength. Reduced and nonreduced sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed peptide structural changes that occurred due to processing, especially for whey proteins. Results are discussed in terms of potential for application of extruded or toasted MPC80 in high-protein nutrition bar applications.

Matrix metalloproteinase-3 gene polymorphism in renal transplant patients with gingival overgrowth.

BACKGROUND AND OBJECTIVE: Gingival enlargement frequently occurs in transplant patients receiving immunosuppressive drugs. It was hypothesized that gingival enlargement associated with cyclosporine use results from reduced degradation of extracellular matrix in the gingiva. Matrix metalloproteinase-3 (MMP-3) is involved in biodegradation of the extracellular matrix, and its inhibition may contribute to an abnormal accumulation of fibronectin and proteoglycans, which are MMP-3 substrates. The aim of this study was to investigate whether an association exists between MMP-3 genotypes and gingival enlargement in kidney transplant patients medicated with cyclosporine A. MATERIAL AND METHODS: Sixty-four unrelated kidney transplant patients suffering from gingival overgrowth, as well as 111 control transplant patients without gingival overgrowth, were enrolled in the study. Gingival overgrowth was assessed 6 mo after transplantation. During the post-transplant period all patients were given cyclosporine A as a principal immunosuppressive agent. MMP-3 polymorphism was determined using a PCR restriction fragment length polymorphism assay. RESULTS: In kidney transplant patients suffering from gingival overgrowth the mean gingival overgrowth score was 1.35 +/- 0.57, whereas in control subjects the mean gingival overgrowth score was 0.0. The distribution of MMP-3-1178A/dupA alleles among all kidney transplant patients, as well as in the two study subgroups, did not differ significantly from Hardy-Weinberg equilibrium. The frequency of the MMP-3-1171A/A genotype (28.1% for gingival overgrowth vs. 26.1% for controls) and of the MMP-3-1171dupA/dupA genotype (32.8% for gingival overgrowth vs. 22.5% for controls) was similar for both study groups. The risk of gingival overgrowth was lowest among patients carrying the MMP-3-1171A/dupA genotype (odds ratio 0.52), but this did not differ markedly from the other genotypes. CONCLUSION: No association between MMP-3 gene polymorphism and gingival overgrowth was revealed in kidney transplant patients administered cyclosporine A.

SPARC gene polymorphism in renal transplant patients with gingival overgrowth.

BACKGROUND: Gingival enlargement frequently occurs in transplant patients receiving immunosuppressive drugs. It was hypothesized that gingival enlargement associated with cyclosporin use results from increases in the number of fibroblasts and the volume of extracellular matrix. SPARC (secreted protein, acidic, and rich in cysteine) regulates cell-matrix interactions, binding to structural matrix proteins, and is induced by cyclosporin A (CsA). The aim of the study was to determine whether there is an association between SPARC genotypes and gingival enlargement in kidney transplant patients given CsA. METHODS: Sixty-two unrelated kidney transplant patients with gingival overgrowth and 124 control transplant patients without overgrowth were enrolled into the study. Gingival overgrowth was assessed at 6 months after transplantation. All patients were given CsA as a principal immunosuppressive agent during the post-transplant period. SPARC polymorphism was determined using polymerase chain reaction-restriction fragments length polymorphism assay. RESULTS: In kidney transplant patients with gingival overgrowth, the mean score of gingival overgrowth was 1.42+/-0.63, whereas in control subjects it was 0. The distribution of SPARC 998C>G alleles among all kidney transplant patients, as well as in the two study subgroups, did not differ significantly from Hardy-Weinberg equilibrium. The frequencies of the 998G allele (24.2% versus 18.5%) and of 998G allele carriers (40.3% versus 33.1%) among individuals with gingival overgrowth was higher compared to the control group, but the differences did not reach the statistical difference. The risk for gingival overgrowth was highest among patients carrying the 998GG genotype (OR 2.25), but it did not differ significantly from the risks associated with the other genotypes. CONCLUSION: No association between SPARC gene polymorphism and gingival overgrowth was revealed in kidney transplant patients who were administered CsA.

Polymorphism in interleukin-1beta gene and the risk of periodontitis in a Polish population.

PURPOSE: The aim of the present study was to explore an association between IL-1B polymorphism and periodontal disease in patients with chronic periodontitis and subjects with aggressive periodontitis in a Polish population. In multivariate logistic regression the association of the following parameters: genotype, age, sex, smoking status, and approximal space plaque index (API) > 50% with the risk of periodontitis was analyzed. MATERIAL AND METHODS: Fifty-two unrelated patients suffering from periodontitis, 20 of them with generalized aggressive periodontitis and 32 with generalized advanced chronic periodontitis were enrolled into the study. Control group consisted of 52 healthy volunteers, without signs of periodontitis. IL-1B(+3954) polymorphism was determined using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: There were no significant differences in the distribution of IL-1B(+3954) genotypes and alleles between periodontal patients either with chronic or aggressive periodontitis and the controls. A predisposing genotype consisting of allele 2 was carried by 34.4% of subjects with chronic periodontitis, 25.0% of subjects with aggressive periodontitis, and 40.3% of healthy subjects. Multivariate logistic regression analysis revealed significant association of age (p = 0.003), smoking (p =0.03), and API > 50% (p = 0.002) with the appearance of aggressive periodontitis, as well as API > 50% (p < 0.001) with chronic periodontitis. CONCLUSIONS: The study revealed no association of IL-1B polymorphism and the risk of aggressive and chronic periodontitis. The risk of aggressive periodontitis was significantly associated with age, smoking, and oral hygiene where as chronic periodontitis with oral hygiene only.

Autogenous bone and platelet-rich plasma (PRP) in the treatment of intrabony defects.

PURPOSE: Obtaining an answer to the question whether autogenous bone in combination with PRP give a therapeutical effect in the form of periodontal ligament attachment regeneration. Defining the degree of elimination of a convenient environment for subgingival bacterial plaque by reduction of periodontal pocket depth and periodontitis. MATERIAL AND METHODS: Twenty-six systematically healthy patients with diagnosed chronic and advanced periodontitis (24 females and 2 males) were selected for the study. In general 72 periodontal infrabony pockets were treated. Clinically the following indexes were examined and measured: 1. Plaque Index by Silness and Löe. 2. Sulcus Bleeding Index by Mühlemann and Son. 3. Clinical Attachment Level (mm). 4. Pocket Depth (mm). 5. Gingival Recession (mm). 6. Tooth mobility with the use of Periotest. 7. Degree of alveolar bone loss with the use of Engelberger, Marthaler and Rateitschak index - EMR Index. RESULTS: At 12 months after treatment the following results were noted: Mean value of attachment level regeneration 3.47 mm. Mean value of pocket depth decreased by 3.7 mm. Mean value of tooth mobility reduction by 48.3%. Megeneration of alveolar bone by 9.24%. CONCLUSIONS: 1. Autogenous bone with added PRP in treatment of intrabony defects caused by periodontitis have given significant clinical improvement of the periodontal tissues. 2. The combination of PRP and autogenous bone caused the elimination of a convenient environment for subgingival bacterial plaque eliminating periodontitis.

Local argyrosis of oral mucosa or amalgam tattoo. A problem in diagnosis and treatment.

The authors, basing on three cases published by different authors in the years 1995-2003, discuss the problem of diagnosis and treatment of local gingival argyrosis and amalgam tattoo. Treatment methods carried out consisted of the following procedures free gingival graft, subepithelial connective tissue graft in a two-step procedure and subepithelial connective tissue graft without flap coverage. In the authors opinion in some cases a connective tissue graft does not need flap coverage, therefore a dual blood supply is not necessary.

Interleukin-6 gene polymorphism in renal transplant patients with and without gingival overgrowth.

OBJECTIVE: To determine whether there is an association between genotypes of interleukin-6 (IL-6) and gingival overgrowth in kidney transplant patients. METHODS: Sixty-three unrelated kidney transplant patients suffering from gingival overgrowth as well 125 control transplant patients without overgrowth were enrolled into the study. Gingival overgrowth was assessed by two independent periodontal specialists at 6 months after transplantation. During the post-transplant period, all patients were given medication, which included cyclosporin A, diltiazem or verapamil, prednisone, and azathioprine. IL-6 polymorphism was determined using the polymerase chain reaction-restriction fragment length polymorphism assay. RESULTS: In kidney transplant patients suffering from gingival overgrowth mean score of gingival overgrowth was 1.41+/-0.64, whereas in control subjects it was 0.0. Patients with gingival overgrowth induced by immunosuppressive medication were characterized by genotypes similar to the controls distribution of IL-6. There were no significant differences of analyzed genotypes' distribution, i.e. -174G/G, -174G/C and -174C/C between patients with gingival overgrowth 33.3%, 39.7%, 27.0% and without gingival overgrowth 30.4%, 49.6% and 20.0%, respectively. The risk of gingival overgrowth was the highest among patients carrying -174C/C genotype (OR 1.48), but did not differ markedly from the other genotypes, i.e. -174G/G (OR 1.15) and -74G/C (OR 0.67). Similar to genotypes, the distribution of alleles was similar in patients with gingival overgrowth and healthy gingiva. The -174G allele was found in 53.2% and 46.8% of subjects whereas -174C allele was revealed in 46.8% and 44.8% of patients with and without gingival overgrowth, respectively. The evaluated risk of gingival overgrowth in patients with -174G allele was 1.09 versus those with healthy gingiva. The medication regimen administered in both groups of the study was comparable. CONCLUSION: No association between the IL-6 gene polymorphism and gingival overgrowth was revealed in kidney transplant patients administered cyclosporin A as a principal immunosuppressive agent.

P-glycoprotein drug transporter MDR1 gene polymorphism in renal transplant patients with and without gingival overgrowth.

OBJECTIVE: To determine whether there is association between genotypes of drug transporter multidrug resistant (MDR)1 gene coding drug transporter P-glycoprotein and gingival overgrowth in kidney transplant patients. METHODS: Fifty-four unrelated kidney transplant patients suffering from gingival overgrowth as well 120 control transplant patients without overgrowth were enrolled into the study. Gingival overgrowth was assessed by two independent periodontal specialists at 6 months after transplantation. During the post-transplant period all patients were given medication, which included cyclosporine A, diltiazem or verapamil, prednisone, azathioprine. MDR1 C3435T polymorphism was determined using the polymerase chain reaction-restriction fragment length polymorphism assay. RESULTS: In kidney transplant patients suffering from gingival overgrowth mean score of gingival overgrowth was 1.43 +/- 0.63, whereas in control subjects was 0.0. Patients with gingival overgrowth induced by immunosuppressive medication were characterized by similar distribution of MDR1 genotypes. There were no significant differences of 3435CC, 20.4% and 22.5%, 3435CT, 61.1% and 54.2% and 3435TT, 18.5% and 23.3% genotypes (frequencies) between patients with and without gingival overgrowth. The risk of gingival overgrowth was the highest among patients carrying 3435CT genotype (OD 1.33), but did not differ markedly from the other genotypes, i.e. 3435CC (OD 0.88) and 3435TT (OD 0.75). Likewise to genotypes, distribution of alleles was similar in patients with gingival overgrowth and healthy gingiva. The wild-type allele 3435C was found in 50.9% and 49.6% of subjects whereas the mutated allele 3435T was revealed in 49.1% and 50.4% of patients with and without gingival overgrowth, respectively. The evaluated risk of gingival overgrowth in patients with 3435C allele was 1.06 versus 0.95 in those with healthy gingiva. The medication regimen administered in both groups of the study was comparable. Immunohistochemical studies revealed expression of P-glycoprotein in ducts of the salivary gland. CONCLUSION: No association between the MDR1 gene polymorphism and gingival overgrowth was revealed in kidney transplant patients administered cyclosporine A as a principal immunosuppressive agent. Further studies are needed to elucidate the role of P-glycoprotein in drug transport in salivary glands.