Evaluation of surface energy and surface stability and adherence of Candida albicans to octa fluoro pentyl (meth) acrylate-coated PEEK using plasma spray.

Background: Polyetheretherketone (PEEK) has favorable properties that make it able to be used as a denture base material, but it is also susceptible to the adhesion of microorganisms. In this study, we applied Octafluoropentyl (meth) acrylate (OFPA) coating on the PEEK polymer surface by using plasma spray and investigated the functional groups present on the surface, changes in the surface energy and Candida albicans adhesion. Materials and Methods: In this experimental study, the samples were placed in a control group without surface preparation and three experimental groups that were subjected to plasma spray for 10, 30, and 60 s and then impregnated with degassed Octa fluoropentyl (meth) acrylate (Sigma-Aldrich, USA) monomer. Fourier transform infrared spectroscopy (FTIR) was used to identify the functional groups and new chemical bonds between PEEK and OFPA, and Sessile Drop Method was used to evaluate the surface's wettability. The surface morphology was checked using a LEXT OLS4000 (Olympus®-Japan) microscope, and the inhibition of C. albicans adhesion was also checked by counting the colonies in terms of colony forming unit/mL (CFU/mL). Kurskal-Wallis analysis was conducted to assess Candida adhesion, while wettability was evaluated using analysis of variance and post hoc analyses. The level of statistical significance was set at P < 0.05. Results: FTIR analysis confirmed that a chemical between OFPA and PEEK was established. The samples showed a significant increase in the contact angle after 30 s of plasma application (CA = 88.2 ± 7.3). The contact angle decreased again by increasing the surface modification to 60 s (CA = 64.33 ± 5.5). Examining the surface morphology of the samples shows an increase in surface roughness with increasing plasma time up to 60 s. The number of adherent colonies was the lowest in 30 s group, but it was not statistically significant (P = 0.658). Conclusion: No statistically significant difference in C. albicans CFU/mL count was found between groups. The contact angle of the 30 s group was significantly higher than the control group.

The application of selenium nanoparticles for enhancing the efficacy of photodynamic inactivation of planktonic communities and the biofilm of Streptococcus mutans.

OBJECTIVE: Streptococcus mutans is one of the principal causative agents of dental caries (tooth decay) found in the oral cavity. Therefore, this study investigates whether selenium nanoparticles (SeNPs) enhance the efficacy of photodynamic therapy (PDT) against both planktonic communities and the one-day-old biofilm of S. mutans. In this study, the planktonic and 24-h biofilm of S. mutans have been prepared in 96-cell microplates. These forms were treated by methylene blue (MB) and SeNPs and then were exposed to light-emitting diode (LED) lighting. Finally, the results have been reported as CFU/ml. RESULTS: The outcomes demonstrated that MB-induced PDT and SeNPs significantly reduced the number of planktonic bacteria (P-value < 0.001). The comparison between the treated and untreated groups showed that combining therapy with SeNPs and PDT remarkably decreased colony-forming units of one-day-old S. mutans biofilm (P-value < 0.05). The findings revealed that PDT modified by SeNPs had a high potential to destroy S. mutans biofilm. This combination therapy showed promising results to overcome oral infection in dental science.

Evaluation of Antibacterial Effects of Cold Atmospheric Plasma, Calcium Hydroxide, and Triple Antibiotic Paste on Enterococcus faecalis Biofilm in the Root Canal System: An In Vitro Study.

Introduction: One of the essential factors in successful endodontic therapy is the effective cleaning and disinfection of the root canal. This study aimed to determine the effect of cold plasma on infected root canals with Enterococcus faecalis and compare its antibacterial effect with the conventional medicaments in vitro. Methods: Sixty-tree single-root teeth were extracted. Canals were cleaned and shaped. Ten teeth were selected as the negative control randomly. The rest of the teeth were incubated at 37°C for 21 days to form E. faecalis biofilm. The specimens were divided into six groups; each group had 10 teeth. In group 1 (the positive control group of calcium hydroxide and triple antibiotic paste [TAP]), methylcellulose was placed in the root canal; in group 2, calcium hydroxide was placed in the root canal for 12 days; in group 3, 10 mg/mL of TAP was placed in the root canal for 12 days; in group 4, helium/oxygen plasma jet was used for 10 minutes. Group 5 was considered as a positive control of plasma, and group 6 was the negative control. After treatment, F4 Pro-Taper rotary file was used to collect root canal microbial biofilms. Bacterial suspensions were serially diluted, and the percentage of growth reduction for each group was obtained by dividing the logarithm of CFU/mL of each group by CFU/mL of the control of the same group. Results: The CFU/mL of TAP and plasma-treated samples was significantly lower than that of the control groups; however, there were no significant differences between the control group and the samples treated by calcium hydroxide. The most percentage of CFU reduction was in the TAP-treated group compared with plasma and calcium hydroxide-treated groups. Conclusion: The application of cold plasma effectively inhibited the growth of E. faecalis and reduced bacterial biofilm. Also, in the present study, 10 mg/mL of TAP caused the complete elimination of E. faecalis. Calcium hydroxide had the most negligible effect on E. faecalis biofilm elimination.

Antimicrobial effects of selenium nanoparticles in combination with photodynamic therapy against Enterococcus faecalis biofilm.

BACKGROUND: Selenium Nanoparticles (SeNPs) were reported as an agent that may enhance the effectiveness of Photodynamic Antimicrobial Chemotherapy (PACT). This in vitro study evaluates the effect of SeNPs on the efficacy of Methylene Blue (MB)-induced PACT against the biofilm formated in 96-well plates and the dentine tubule biofilm of Enterococcus faecalis. METHODS: Chitosan coated SeNPs were synthesized using chemical reduction method and were characterized by Transmission Electron Microscope (TEM) and Dynamic Light Scattering (DLS). Twenty-four-hour biofilms of E. faecalis were developed on 96-well plates and treated with SeNPs, MB, and Light-Emitting Diode (LED). Also, three-week biofilms of E. faecalis were formed on 67 specimens of dentinal tubules, and the antibacterial effects of MB+SeNPs on these biofilms were studied. RESULTS: The average hydrodynamic diameter of SeNPs was 80/3 nm according to DLS measurement. The combined use of MB and SeNPs significantly reduced Colony-Forming Units (CFUs) of one-day-old E. faecalis biofilms in comparison with the control group (P value < 0.05). Besides, combination therapy had the most antibacterial effect on root canal E. faecalis biofilms at both 200 and 400 µm depths of dentine tubules (P value < 0.001). Of note, about 50% of human fibroblast cells survived at a concentration of 128 µg/ml of SeNPs, compared to the control group. CONCLUSION: The results demonstrated that the photodynamic therapy modified by SeNPs could be an effective disinfection alternative to the destruction of E. faecalis biofilms and root canal treatment.

Effect of Apical Size and Taper on the Efficacy of Root Canal Disinfection With LED Photodynamic Therapy as an Adjunct to Irrigation With Sodium Hypochlorite.

Introduction: This study assessed the effect of apical size and taper on the efficacy of root canal disinfection with LED photodynamic therapy (PDT) as an adjunct to irrigation with sodium hypochlorite. Methods: A total of 126 extracted human mandibular molars were divided into 4 groups. The mesiobuccal canal was prepared to size 25/4% in group 1, 25/6% in group 2, 30/4% in group 3, and 30/6% in group 4 using the iRaCe rotary system. A 21-day Enterococcus faecalis biofilm was prepared and used for inoculation of the canals. Each group was randomly divided into 3 subgroups for canal disinfection with 2.5% sodium hypochlorite, sodium hypochlorite plus LED PDT and saline (positive control). Samples from the root canals were obtained with rotary files and cultured. Microbiologic data were analyzed using the Poisson regression test. Results: The bacterial count significantly decreased following disinfection with sodium hypochlorite with/without PDT in all sizes and tapers of preparation compared with the control group (P<0.05). Increasing the apical taper or apical size and the use of PDT as an adjunct did not have a significant effect on the reduction of the bacterial count (P>0.05). However, the apical size and PDT had a significant effect on the number of residual bacteria (P<0.05), and increasing the apical size and conduction of PDT significantly decreased the number of residual bacteria. Conclusion: The apical size and taper and the use of PDT as an adjunct did not have a significant effect on the reduction of the bacterial count. However, increasing the apical size and conduction of PDT as an adjunct to sodium hypochlorite irrigation significantly decreased the number of residual bacteria in the root canal system.

The comparison of the salivary concentration of interleukin-17 and interleukin-18 in patients with chronic periodontitis and healthy individuals.

BACKGROUND: Regarding the prevalence and importance of periodontal disease and the potential of salivary biomarkers for the early diagnosis of these diseases, this study was conducted to compare salivary concentrations of Interleukin-17 (IL-17) and Interleukin-18 (IL-18) in patients with chronic periodontitis and healthy individuals. MATERIALS AND METHODS: The present research was a descriptive-analytical and also a cross-sectional study. Unstimulated saliva with full-mouth clinical periodontal recordings were obtained from 20 healthy individuals and 20 individuals with chronic periodontitis. The concentrations of salivary IL-17 and IL-18 were determined using the enzyme-linked immunosorbent assays. The nonparametric Mann-Whitney U-test was used for statistical analysis of the findings. Alpha level was set at 0.05. RESULTS: The mean salivary concentration of IL-18 in patients with chronic periodontitis was 143.10 pg/mL, which was higher than the same concentration in healthy controls (78.33 pg/mL), (P = 0.035). The mean salivary concentration of IL-17 in patients with chronic periodontitis and healthy controls was 3.51 and 4.57 pg/mL, respectively, and there was no difference between the two groups (P = 0.283). CONCLUSION: Within the limitations of the present study, it may be suggested that an elevated salivary IL-18 level in chronic periodontitis patients has the potential to be a biomarker for periodontal tissue destruction.

Secretor Status of ABO Antigens in Saliva of a Defined Group of Iranian Patients with Pemphigus Vulgaris: A Case-Control Study.

INTRODUCTION: Pemphigus is a chronic inflammatory and autoimmune disease which can cause blisters and mucocutaneous erosions. ABO secretor refers to those who secrete ABO blood group antigens based on their blood type in body fluids such as saliva, sweat, tears, semen, and serum. Previous studies showed that nonsecretor people are more prone to certain autoimmune diseases. AIM: The aim of this study was to determine the ABO secretor status in the saliva of patients with pemphigus vulgaris. MATERIALS AND METHODS: This case-control study was conducted on 35 patients with pemphigus vulgaris and 35 healthy controls. The two groups were matched for age and gender. Pemphigus vulgaris diagnosis was confirmed by histopathology and direct immunofluorescence microscopy. ABO blood grouping was done, and 5 ml of unstimulated saliva was collected to determine secretor status. Secretors were recognized from nonsecretors by the Wiener agglutination inhibition test. Results were extracted by using statistical chi-square and Fisher's exact tests. RESULTS: 16 male and 19 female patients aged 49.43 ± .12.37 years were compared with 16 male and 19 female controls aged 46.43 ± 11.88 years. The most frequent blood group among case and control groups was O (54.3% and 60%, respectively). There was no significant difference in blood groups (P=0.73). 90% of the samples were ABO secretors. The patient group included 31 (88.6%) and the control group included 32 (91.4%) ABO secretors; there was no significant difference between the two groups (P=1.000). CONCLUSION: In this study, we observed that the people with nonsecretor status in comparison with the people with secretor status are not more susceptible to develop pemphigus vulgaris.

Comparison of osteopromoting ability of human tooth powder with the demineralized freeze-dried bone allograft, a bovine xenograft, and a synthetic graft: An in vitro study.

Background: The present study aimed to evaluate the osteopromoting ability of human tooth powder and compare it to a bovine xenograft, a synthetic material, and the DFDBA allograft. Methods: In this in vitro study, 30 teeth without caries, inflammation, and infection, which had been extracted for orthodontic reasons, were collected. The crowns were removed, pulpectomy was carried out, and the samples were ground to a powder with particles <500 µm. Osteoblast-like cells of MG-63 were cultured with the tooth powder, Cerabone, DFDBA, and Osteon II. Cell proliferation was assessed by the MTT assay at 24- and 72-hour intervals. The alizarin red test was carried out after three and five days. The alkaline phosphatase level was measured after 24, 48, and 72 hours to assess the osteoblastic activity. The results were analyzed with one-way ANOVA. Results: According to the MTT assay, all the materials exhibited a higher proliferation rate than the control group in 24 hours. In 72 hours, DFDBA had the lowest cell proliferation rate at concentrations of 40 and 80 mg/mL. DFDBA and the positive control group were able to create calcified nodules by the alizarin red test. At the 48- and 72-hour intervals, DFDBA had the lowest alkaline phosphatase activity at a concentration of 40 mg/mL. At the 72-hour interval, bovine xenograft had the highest alkaline phosphatase level, followed by the synthetic material and tooth powder. Conclusion: The tooth powder was able to increase cell proliferation in comparison with the bovine xenograft, the synthetic graft, and the DFDBA. However, its osteopromoting ability was less than that of the osteogenic materials.

Does Melissa officinalis consumption improve the salivary antioxidant status of smokers?

BACKGROUND: The aim of this study was to investigate the effects of Melissa officinalis tea on the total antioxidant capacity of saliva among smokers. METHODS: 24 smokers were selected by convenience sampling. Demographic information and duration of smoking were recorded at the beginning of study. Two cups of Melissa officinalis tea were given to the participants with specific instruction for 30 days. The unstimulated saliva was collected on first day, 15th and 30th days. Then, total antioxidant capacity was measured by a special kit. Statistical analysis was conducted by repeated measure ANOVA test. RESULTS: The mean values of total antioxidant capacity of saliva were significantly higher in days 15 and 30 from the baseline. (p<0.0001, P=0.006). In day 30, the mean value of antioxidant was not significantly different from day 15. (P=0.271). CONCLUSION: Melissa officinalis tea consumption increases salivary antioxidants level in smokers.

Molecular characterization of enterotoxigenic Escherichia coli isolated from neonatal calves diarrhea.

Enterotoxigenic Escherichia coli (ETEC) produces different virulence factors allowing the bacterium to colonize and develop watery diarrhea. Proteomics studies have also introduced new protein belonging to the secretion pathways, antigen 43 (Ag43), which plays important role in E. coli pathogenesis. The objective of this study was to investigate O-types and virulence factors of E. coli isolates from neonatal calves diarrhea. Total of 120 isolates from diarrheic calves were genotyped for their O groups and the presence of virulence genes K99, F41 and STa as well as Ag43. The predominant O-type was O101 (51.00%) and the prevalence of K99, F41 and STa was 7 (5.80%). The Ag43 was detected in all samples with three different allelic patterns. Our results indicated that K99 positive isolates certainly have one of each 2200 bp or 1800 bp or both copies of Ag43 passenger domain, while negative K99 isolates lack the Ag43. The results reported here provide informative data regarding the prevalence of E. coli O-types and their virulence factors in enteric colibacillosis. The Ag43 that was more found in K99 positive isolates might be associated with diarrhea-causing E. coli strains in neonatal calves.

Antimicrobial efficacy of clindamycin and triple antibiotic paste as root canal medicaments on tubular infection: An in vitro study.

This study compared the antibacterial effect of 2% clindamycin and 2% and 100% concentration of triple antibiotic paste (TAP) on an Enterococcus faecalis biofilm. Dentinal tubules of 100 root specimens were infected and randomly assigned to five groups. A total of 1000 mg mL-1 of TAP, 20 mg mL-1 of TAP and clindamycin, calcium hydroxide or methylcellulose (control) were placed in the root canal for 1 week. After treatment, dentine shavings were collected from 200 and 400 µm dentine depth and the number of colony-forming units (CFU) per mg was determined. Reduction in viable bacteria in first three groups was significantly better than calcium hydroxide and control groups. However, the antimicrobial effectiveness among these three groups was not significantly different from each other. There was no significant difference between data at 200 and 400 µm in all groups except the Ca(OH)2 group. The antibiofilm effect of clindamycin was comparable with TAP, so it may be used instead of TAP.

Evaluation of Resistin Levels in Saliva of Patients with Chronic Periodontitis and Healthy Subjects.

OBJECTIVE: To evaluate resistin levels in the saliva of patients with chronic periodontitis, and healthy subjects. METHODS: Thirty-four subjects aged between 25 and 50 years were included and divided into healthy group (n = 19) and chronic periodontitis group (n = 15). The saliva levels of resistin were assessed by enzyme-linked immunosorbent assay. Comparisons of resistin levels between the two groups were made with the Mann-Whitney Test. RESULTS: The chronic periodontitis group showed significantly higher resistin levels than the control group (P = 0.001). CONCLUSION: The level of resistin in saliva might help to determine the inflammatory status of periodontal diseases.

Periostin levels in saliva of patients with chronic periodontitis.

BACKGROUND: Periostin acts as a necessary protein for tissue integrity and maturity and has a key role as a modulator of periodontal ligament hemostasis. It has been shown that periostin acts as a supportive protein. The aim of this study was to compare the concentration of periostin in the saliva of patients with chronic periodontitis and healthy controls. MATERIALS AND METHODS: In this case-control, cross-sectional study, a total of 45 individuals (25 patients with chronic periodontitis and 20 healthy controls) were evaluated. Whole saliva samples were collected, and periostin levels were evaluated by standard enzyme-linked immunosorbent assay. The results were analyzed by SPSS and Mann-Whitney analysis. RESULTS: The results of this study showed that the level of periostin in saliva in patients with periodontitis was significantly lower than healthy controls (P < 0.05). Periostin was detectable in all samples. CONCLUSION: The results show that there is a significant relationship between the level of periostin in saliva and chronic periodontitis. Periostin may be considered as an inflammatory marker in periodontal disease. However, further studies are needed to confirm this finding.

Gingival crevicular fluid levels of sclerostin in chronic periodontitis and healthy subjects.

OBJECTIVES: Chronic periodontitis is a common inflammatory disease of the oral cavity that causes destruction of periodontal tissues and bone around the teeth. Sclerostin is a protein encoded by the SOST gene. In this study, gingival crevicular fluid (GCF) levels of sclerostin in patients with chronic periodontitis were compared with those of healthy subjects. MATERIALS AND METHODS: In this case-control study, a total of 40 subjects were enrolled and divided into the healthy group (n=23) and chronic periodontitis group (n=17). GCF samples were collected, and the concentration of sclerostin was evaluated using enzyme-linked immunosorbent assay. Comparison of significance between groups was assessed using Mann-Whitney U test. RESULTS: Sclerostin concentration was significantly higher in the chronic periodontitis group compared with the healthy group (P<0.005). CONCLUSION: Despite the limitations of this study, sclerostin can be a possible marker for assessment of periodontal health status.

Comparison of Attachment and Proliferation of Human Gingival Fibroblasts on Different Collagen Membranes.

BACKGROUND AND AIM: Human gingival fibroblasts cultured on collagen membrane as an alternative treatment method used in tissue regeneration can lead to improved results in root coverage. The aim of this study was to evaluate the human gingival fibroblast proliferation and adhesion cultured on three types of collagen membranes. MATERIALS AND METHODS: In this in vitro study, first-line human gingival fibroblast cells (HGF1-RT1) prepared and cultured on three membranes, including porcine pericardium (PP) (Jason, Botiss dental), human pericardium (HP) (Regen, Faravardeh Baft Iranian), and glutaraldehyde cross-linked (GC) (BioMend Extend, Zimmer Dental). Cell survival was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) after 24, 48, and 72 h and 7 days. Furthermore, morphology and adhesion of cells on the membrane were evaluated after 1 and 7 days by electron microscopy (scanning electron microscopy [SEM]). Statistical analysis was performed using two-way ANOVA with a significance level of 0.05. RESULTS: Based on the results of MTT, cell survival on HP and PP membranes after 7 days significantly increased (P < 0.001), but for the GC membrane, it was reduced after 7 days (P = 0.031). Cell survival on HP and PP membranes did not differ (P = 1) and was more than GC (P < 0.001). SEM images showed that the adhesion of cells was better on HP and PP membranes than GC. CONCLUSION: The results of this study showed that natural collagen membranes (HP and PP) similarly support proliferation and adhesion of gingival fibroblasts. Survival and adhesion of gingival fibroblasts on cross-linked collagen membrane was less than two other membranes.

Correction to: Comparison of the effect of activated or non-activated PRP in various concentrations on osteoblast and fibroblast cell line proliferation.

In the original publication of this article, the affiliation of the corresponding author has been published incorrectly. Now the correct affiliation has been provided in this erratum.

In Vitro Cytotoxicity and Setting Time Assessment of Calcium-Enriched Mixture Cement, Retro Mineral Trioxide Aggregate and Mineral Trioxide Aggregate.

INTRODUCTION: The present study sought to evaluate and compare biocompatibility and setting time of Retro mineral trioxide aggregate (MTA), calcium-enriched mixture (CEM) and Angelus MTA. METHODS AND MATERIALS: CEM cement, Angelus MTA and Retro MTA were assessed in set and fresh states. Extracts transformed to each cavity of three 24-well plates in which 1×104 cell were seeded into each well 24 h earlier. All specimens were incubated in a humidified incubator with 5% CO2 at 37°C. Mosmann's tetrazolium toxicity (MTT) assay was used to determine in vitro cytotoxicity on L929 mouse fibroblast cell line. Cell viability was determined at 1, 24, and 72 h after exposure. The initial setting time was measured by 113.4 g Gilmore needle testing. Then, final setting times were assessed by the 456.5 g Gilmore needle. Data comparisons were performed using the analysis of variance (ANOVA) and Tukey's post hoc test (α=0.05). RESULTS: All groups in both forms indicated higher cell vitality compared to positive control group (P<0.001). After 24 h, the set Retro MTA showed better biocompatibility compared to set CEM and set Angelus MTA (P<0.001). Retro MTA showed significantly lower initial and final setting time compared to CEM and Angelus MTA (P<0.001). CONCLUSION: Our results indicated the good cell viability values of Retro MTA and relatively short period of setting time. It seems a promising alternative material in clinical situations where accelerated setting is required. However, more clinical and in vivo investigations are needed for a clear decision making.

Comparison of the effect of activated or non-activated PRP in various concentrations on osteoblast and fibroblast cell line proliferation.

Platelet-rich plasma (PRP) contains growth factors which positively affect cell proliferation, cell differentiation, chemotaxis and intracellular matrix synthesis. All these processes are involved in wound healing and tissue regeneration; thus, PRP as a source of growth factors can be used in periodontal regenerative therapies. The purpose of the present study was to assess the effect of various concentrations of activated and non-activated PRP on proliferation of osteoblasts and fibroblasts in vitro. PRP was obtained from three healthy volunteers. 75, 50, 25, and 10% concentrations of f PRP were prepared by dilution in Dulbecco's modified Eagle's medium. In activated PRP groups, PRP concentrations were activated by adding calcium gluconate. Human gingival fibroblast (HGF) cell line and MG-63 (osteosarcoma) human osteoblast-like cell line were used in the study. The MTT proliferation assay was used to assess the effect of different types of PRP concentrates on proliferation of HGF and MG-63 cells, in 24, 48 and 72 h. After 24, 48, and 72 h, the proliferation rate of both cell lines was higher in the positive control group, except in 72 h in HGF cell lines, that 10% non-activated PRP group and 10 and 25% activated PRP groups has higher proliferation rate than the positive control group, which it was not significant. Proliferation rate in cells with 10% activated PRP was highest among samples containing PRP. The current study failed to show the significant effect of activated or non-activated PRP on proliferation of HGFs or MG-63 osteoblast-like cells. However, our results showed that activated PRP had a greater effect than non-activated PRP.

Expression of Recombinant Human Amelogenin in Iranian Lizard Leishmania and Its Biological Function Assay.

BACKGROUND: Amelogenins are the major components of enamel matrix proteins. Enamel matrix derivatives (EMD) can be used in periodontal diseases to regenerate periodontal tissues. The main aim of this study was to evaluate expression of full-length functional recombinant human amelogenin (rhAm) in Iranian lizard Leishmania (I.L.L.) as an alternative eukaryotic expression system. METHODS: Human cDNA encoding a 175-amino acid amelogenin expression cassette was sub cloned into a pLEXSY vector. The construct was transferred into Leishmania cells by electroporation. The protein production was surveyed in the transcription and the translation levels. The expressed protein was purified and some of its biological properties were investigated in comparison to EMD and negative control. RESULTS: Expression of rhAm was confirmed by RT-PCR and western blot test in Leishmania cells. Purified rhAm significantly inhibited the formation of tartrate-resistant acid phosphatase positive (TRAP(+)) multinuclear cells in calcitriol stimulated mouse marrow cultures. Moreover, it significantly promoted proliferation and DNA synthesis in L929 mouse fibroblast cells. CONCLUSION: Functional rhAm was successfully expressed in I.L.L. Easy handling and post translation modification were the main advantages of this expression system. It is suggested to investigate molecular properties of this rhAm in the future.