Isolation, characterization and regulation of moonlighting proteases from Candida glabrata cell wall

Candida glabrata (C. glabrata) cell wall proteins play a role in virulence and in initial host immune recognition and responses. We isolated and characterized C. glabrata cell wall proteases from a clinical hospital C. glabrata T1638 blood isolate and estimated the enzymatic activities and their ability to degrade gelatin and processing proMMP-8 and assess the regulation of these proteases with salt treatment, mercaptoethanol and fermented lingonberry juice from Vaccinium vitis idaea L. The cell wall proteases were enzymatically released from the cell wall and beta 1,3bonded proteases were fractioned into 10-50 kDa and >50 kDa fractions with anionic DEAEsepharose ion-exchange chromatography and gel filtration. Proteins were monitored and analyzed with MDPFzymography, and five gelatinolytic bands were cut out from a parallel silver-stained gel for the LCMS/MS analysis. The proteases lacked a signal sequence, indicating that they are moonlighting proteases. Human proMMP-8 activation assays were performed with both fractions and verified by western-immunoblot using aMMP-8 specific antibody. Inhibition of proMMP-8 conversion to the lower molecular active enzyme species were demonstrated with fermented lingonberry juice. The results indicate that moonlighting proteases may play a role in the virulence of C. glabrata.Peer reviewe

Matrix metalloproteinase MMP-8, TIMP-1 and MMP-8/TIMP-1 ratio in plasma in methicillin-sensitive Staphylococcus aureus bacteremia

Background Matrix metalloproteinase-8 (MMP-8) and tissue inhibitor of metalloproteinases-1 (TIMP-1) have been shown to predict prognosis in sepsis. However, MMP-8 and TIMP-1 in Staphylococcus aureus bacteremia (SAB) lacks evaluation and their role in the pathogenesis of SAB is unclear. Methods MMP-8 and TIMP-1 and MMP-8/TIMP-1 molar ratio were determined at days 3, 5 and 28 from positive blood cultures in patients with methicillin-sensitive SAB and the connection to disease severity and early mortality was determined. Results Altogether 395 SAB patients were included. Patients with severe sepsis or infection focus presented higher MMP-8 levels at day 3 and 5 (p Conclusion MMP-8 and MMP-8/TIMP-1 ratio were high 3-5 days after MS-SAB diagnosis in patients with an infection focus, severe sepsis or mortality within 14 days suggesting that matrix metalloproteinase activation might play a role in severe SAB.Peer reviewe

Lingonberry polyphenols : Potential SARS-CoV-2 inhibitors as nutraceutical tools?

Proposed pathway of the effect of lingonberry polyphenols on oral microbial (viral) load reduction and consequent beneficial local and systemic (respiratory tract) anti-inflammatory and antimicrobial/antiviral effects.Non peer reviewe

Effects of Fermented Lingonberry Juice Mouthwash on Salivary Parameters—A One-Year Prospective Human Intervention Study

A one-year prospective human intervention study was performed to examine the effects of fermented lingonberry juice (FLJ), used as a mouthwash for six months, on salivary parameters. A total of 25 adult participants used 10 mL of FLJ as mouthwash 30 s daily for 6 months in addition to their normal oral homecare routines. Standard oral examinations and gathering of samples were performed at the beginning of the study and after six months and one year. Resting and stimulated saliva secretion rates, resting saliva pH, and stimulated saliva buffering capacity were determined. A questionnaire of participants’ subjective sensations of mouth dryness was also recorded at each timepoint. Fermented lingonberry juice mouthwash had positive effect to all five salivary parameters and were, according to the omnibus test, statistically significant during the study period. Analysis of the subjective dry mouth sensation questionnaires revealed that symptoms of xerostomia decreased due to the use of FLJ. This study revealed that the once-a-day use of FLJ mouthwash had a beneficial, increasing effect on salivary flow rates, buffering capacity, and salivary pH. FLJ thus can be safely used as an adjunctive and beneficial therapy in oral homecare, protecting teeth and oral mucosa, including periodontium, and also relieving dry mouth symptoms

Fermented lingonberry juice's effects on active MMP-8 (aMMP-8), bleeding on probing (BOP), and visible plaque index (VPI) in dental implants-A clinical pilot mouthwash study

Objectives: We aimed to study the effects of fermented lingonberry juice (FLJ) as a mouthwash on the levels of active matrix metalloproteinase-8 (aMMP-8) in peri-implant sulcular fluid (PISF), bleeding on probing (BOP), and visible plaque index (VPI). We hypothesized that FU rinsing could reduce inflammation (aMMP-8 and BOP) and microbial load (VPI) in the oral cavity, especially around dental implants. Materials and Methods: A clinical pilot study was performed using FLJ as a mouthwash. The inclusion criteria were at least one dental implant in the anterior or posterior areas with a screw-retained crown. Ten participants used 10 ml of mouthwash twice a day for 15 days, and 10 participants served as the control group. Point-of-care tests (POCTs) were used to measure aMMP-8 levels in the PISF, and BOP and VPI were recorded at the beginning of the trial and after 15 and 30 days. Results: The FLJ mouthwash had a reductive effect on aMMP-8, VPI, and BOP in the mouthwash group; however, there was no significant difference compared to the control group. The difference in VPI and BOP levels between the groups diminished after the lingonberry regimen ended. The decrease in aMMP-8 levels appeared to continue even after discontinuation of the mouthwash regimen. Conclusion: The reduction in the amount of plaque, aMMP-8, arid BOP by FLJ was promising and continuous considering the relatively short study period and sample size. FLJ is a natural and safe supplement for oral and dental implant home care. Further studies are required to verify these promising results.Peer reviewe

Effects of Fermented Lingonberry Juice Mouthwash on Salivary Parameters-A One-Year Prospective Human Intervention Study

A one-year prospective human intervention study was performed to examine the effects of fermented lingonberry juice (FLJ), used as a mouthwash for six months, on salivary parameters. A total of 25 adult participants used 10 mL of FLJ as mouthwash 30 s daily for 6 months in addition to their normal oral homecare routines. Standard oral examinations and gathering of samples were performed at the beginning of the study and after six months and one year. Resting and stimulated saliva secretion rates, resting saliva pH, and stimulated saliva buffering capacity were determined. A questionnaire of participants' subjective sensations of mouth dryness was also recorded at each timepoint. Fermented lingonberry juice mouthwash had positive effect to all five salivary parameters and were, according to the omnibus test, statistically significant during the study period. Analysis of the subjective dry mouth sensation questionnaires revealed that symptoms of xerostomia decreased due to the use of FLJ. This study revealed that the once-a-day use of FLJ mouthwash had a beneficial, increasing effect on salivary flow rates, buffering capacity, and salivary pH. FLJ thus can be safely used as an adjunctive and beneficial therapy in oral homecare, protecting teeth and oral mucosa, including periodontium, and also relieving dry mouth symptoms.Peer reviewe

Lingonberries—General and Oral Effects on the Microbiome and Inflammation

Lingonberry (Vaccinium vitis ideae L.) is a low-bush wild plant found in the northern hemisphere. The berries are used in traditional medicine in Finland to treat oral yeast infections. General and oral effects of lingonberries on the microbiome and inflammation are reviewed. A brief introduction to oral microbiome symbiosis and dysbiosis, innate and adaptive immunity and inflammation are included, and special features in microbe/host interactions in the oral environment are considered. In vitro anticancer, antimicrobial, antioxidant, anti-inflammatory, and in vivo mouse and human studies are included, focusing on the symbiotic effect of lingonberries on oral and general health

Prediabetes/Diabetes Can Be Screened at the Dental Office by a Low-Cost and Fast Chair-Side/Point-of-Care aMMP-8 Immunotest

Pre-diabetes and diabetes are strongly associated with periodontal disease (gingivitis and periodontitis), and these conditions are known to upregulate aMMP-8 in inflamed gingiva and oral fluids. Thus, it would be feasible to screen for prediabetes and diabetes at the dental office by chairside tests. Chair-side assessment of HbA1c and a quantitative point-of-care (PoC) active matrix metalloproteinase (aMMP)-8 oral rinse immunotest developed for periodontal diseases, were performed on patients (n = 69) attending a Periodontology University Clinic who fulfilled the criteria for testing according to the screening questionnaire of the Centers for Disease Control and Prevention, USA. Clinical parameters of periodontal disease were also recorded with an automated probe. Twenty seven-point-five percent of the subjects were found with previously unknown hyperglycemia (HbA1c ≥ 5.7%). There was a statistically-significant positive association between the aMMP-8test and prediabetes (p < 0.05, unadjusted and adjusted for BMI and age ≥ 45 years logistic regression models). The dental setting is suitable for opportunistic screening for undiagnosed diabetes and pre-diabetes and point-of-care HbA1c, especially when combined with aMMP-8 assessment by dental professionals, being convenient and effective

Lingonberries—General and Oral Effects on the Microbiome and Inflammation

Lingonberry (Vaccinium vitis ideae L.) is a low-bush wild plant found in the northern hemisphere. The berries are used in traditional medicine in Finland to treat oral yeast infections. General and oral effects of lingonberries on the microbiome and inflammation are reviewed. A brief introduction to oral microbiome symbiosis and dysbiosis, innate and adaptive immunity and inflammation are included, and special features in microbe/host interactions in the oral environment are considered. In vitro anticancer, antimicrobial, antioxidant, anti-inflammatory, and in vivo mouse and human studies are included, focusing on the symbiotic effect of lingonberries on oral and general health

Enhanced Systemic Response of Matrix Metalloproteinases and Their Regulators in Campylobacter and Salmonella Patients

Campylobacters are major enteropathogens worldwide with a substantial financial burden. Matrix metalloproteinases (MMPs) are proteolytic metalloendopeptidases with ability to modify immune response and shown to be upregulated in patients with several tissue destructive diseases, including infections. We measured here serum concentrations of MMP-8 and MMP-9 together with their regulators myeloperoxidase (MPO), human neutrophil elastase (HNE), and tissue inhibitor of metalloproteinases (TIMP)-1 in 80 Campylobacter and 25 Salmonella patients as well as in 27 healthy controls. Paired serum samples were available for 73 and 23 patients, respectively. When the initial serum samples were compared to those from controls, both Campylobacter and Salmonella patients showed elevated concentrations of all biomarkers tested (p 0.037). In the follow-up samples, collected about 25 days afterwards, MMP-8 levels of Campylobacter patients had already turned to normal but all the other biomarkers still showed elevated, although from the initial levels significantly dropped, levels. For the follow-up samples of Salmonella patients, only MMP-9 and MPO levels were at a significantly higher level than in controls. It remains to be studied if the systematically enhanced neutrophil-derived proteolytic and oxidative stress, induced by Campylobacter infection as shown here and persisting for several weeks, is important for the development of late sequelae.Peer reviewe