Refermentation with yeast and lactic acid bacteria isolates: a strategy to improve the flavor of green coffee beans.

BACKGROUND: Yeast and lactic acid bacteria (LAB) play an important part in the post-harvest fermentation of coffee. This study applied lab-scale fermentation to commercial green coffee beans using dry coffee pulp as the substrate, with the aim of modifying coffee-bean flavor. In addition to spontaneous fermentation, yeast and LAB isolated from coffee beans and dried coffee pulp were added during fermentation. RESULTS: Co-inoculation of yeast and LAB showed a significant effect on the chlorogenic acid content after between 24 and 72 h of fermentation. Acetic, citric, malic, lactic, and quinic acids were shown to be affected significantly (P < 0.05) by fermentation and inoculation. Gas chromatography detected that esters, alcohols, aldehydes, furans, and pyrazines were the primary compounds in the coffee beans. Certain volatile groups were present in greater concentrations and broader varieties within the inoculated beans. The highest cupping scores were given to beans that had been co-inoculated with yeast and LAB. CONCLUSION: Overall, the use of yeasts and LAB starters showed potential to create coffee beverages with desirable characteristics by standardized fermentation. © 2024 Society of Chemical Industry.

Distribution and quantification of activated platelets in platelet-rich fibrin matrices.

Platelet-rich fibrin (PRF) has been widely applied in regenerative therapy owing to its simple preparation protocol. To date, the original protocol for preparing leukocyte-rich (L)-PRF has been modified to produce derivatives such as advanced (A)-PRF, concentrated growth factors (CGF), and horizontal (H)-PRF. However, these derivatives have not been rigorously compared to explore possible differences. We previously developed and validated a nondestructive near-infrared (NIR) imaging method to quantitatively examine the platelet distribution in PRF matrices. To further evaluate the characteristics of platelets in PRF, we herein examined the distribution of activated platelets. Four types of PRF matrices were prepared under different centrifugal conditions from blood samples obtained from the same healthy donors. After fixation and compression, the matrices were stained immunohistochemically without sectioning and visualized using an NIR imager. Qualitative morphological analysis revealed that whole platelets were distributed widely and homogeneously in H-PRF and A-PRF, but localized along the distal tube surface in L-PRF and CGF. Activated platelets were distributed as were whole platelets in A-PRF, L-PRF, and CGF, but localized mainly in the "buffy coat" region in H-PRF. Quantitative analysis revealed that there was no significant difference in the ratio of activated to whole platelets between PRF derivatives. These findings suggest that platelet activation is similarly induced in fibrin matrices regardless of centrifugal speed or rotor angulation. However, only the H-PRF group was distinguishable from the other PRF derivatives in terms of activated platelet distribution.

Fluorescent Cytochemical Detection of Polyphosphates Associated with Human Platelets.

Polyphosphate (polyP) is released from activated platelets and activates the intrinsic coagulation pathway. However, polyP may also be involved in various pathophysiological functions related to platelets. To clarify these functions, we established a cytochemical method to reproducibly visualize polyP in platelets. Platelets obtained from healthy non-smoking donors were suspended in phosphate-buffered saline and quickly immobilized on glass slides using a Cytospin. After fixation and membrane permeabilization, platelets were treated with 4',6- diamidino-2-phenylindole (DAPI) and examined using a fluorescence microscope with a blue-violet excitation filter block (BV-2A). Fixed platelets were also subjected to immunocytochemical examination to visualize serotonin distribution. Under the optimized conditions for polyP visualization, immobilized platelets were fixed with 10% neutral-buffered formalin for 4 h or longer and treated with DAPI at a concentration of 10 µg/mL in 0.02% saponin- or 0.1% Tween-20-containing Hanks balanced salt solution as a permeabilization buffer for 30 min at room temperature (22-25 °C). Based on the results obtained by using activated platelets, treatment with alkaline phosphatases, and serotonin release, the DAPI+ targets were identified as polyP. Therefore, this cytochemical method is useful for determining the amount and distribution of polyP in platelets.

Fluorometric Quantification of Human Platelet Polyphosphate Using 4',6-Diamidine-2-phenylindole Dihydrochloride: Applications in the Japanese Population.

Polyphosphate (polyP), a biopolymer of inorganic phosphate, is widely distributed in living organisms. In platelets, polyP is released upon activation and plays important roles in coagulation and tissue regeneration. However, the lack of a specific quantification method has delayed the in-depth study of polyP. The fluorescent dye 4',6-diamidine-2-phenylindole dihydrochloride (DAPI) has recently received attention as a promising probe for the visualization and quantification of cellular polyP levels. In this study, we further optimized quantification conditions and applied this protocol in quantification of platelet polyP levels in a Japanese population. Blood samples were collected from non-smoking, healthy Japanese subjects (23 males, 23 females). Washed platelets were fixed and probed with DAPI for fluorometric determination. PolyP levels per platelet count were significantly higher in women than that in men. A moderate negative correlation between age and polyP levels was found in women. Responsiveness to CaCl2 stimulation was also significantly higher in women than that in men. Overall, our optimized protocol requires neither purification nor degradation steps, reducing both the time and bias for reproducible quantification. Thus, we suggest that despite its low specificity, this DAPI-based protocol would be useful in routine laboratory testing to quantify platelet polyP levels efficiently and economically.

Recent advances in ionic liquids-based hybrid processes for CO2 capture and utilization.

CO2 capture and utilization (CCU) is an effective strategy to mitigate global warming. Absorption, adsorption and membranes are methods used for CO2 separation and capture, and various catalytic pathways have also been developed for CO2 utilization. Although widely researched and used in industry, these processes are energy-intensive and this challenge needs to be overcome. To realize further optimization, novel materials and processes are continuously being developed. New generation materials such as ionic liquids (ILs) have shown promising potential for cost-effective CO2 capture and utilization. This study reviews the current status of ILs-based solvents, adsorbents, membranes, catalysts and their hybrid processes for CO2 capture and utilization. The special properties of ILs are integrated into new materials through hybridization, which significantly improves the performance in the process of CCU.

Quantitative Near-Infrared Imaging of Platelets in Platelet-Rich Fibrin (PRF) Matrices: Comparative Analysis of Bio-PRF, Leukocyte-Rich PRF, Advanced-PRF and Concentrated Growth Factors.

Platelet-rich fibrin (PRF) is a fibrin matrix enriched with platelets. The PRF matrix is thought to form a steep gradient of platelet density around the region corresponding to the buffy coat in anticoagulated blood samples. However, this phenomenon has not yet been proven. To visualize platelet distribution in PRF in a non-invasive manner, we utilized near-infrared (NIR) imaging technology. In this study, four types of PRF matrices, bio-PRF, advanced-PRF (A-PRF), leukocyte-rich PRF (L-PRF), and concentrated growth factors (CGF) were compared. Blood samples collected from healthy, non-smoking volunteers were immediately centrifuged using four different protocols in glass tubes. The fixed PRF matrices were sagittally divided into two equal parts, and subjected to modified immunohistochemical examination. After probing with NIR dye-conjugated secondary antibody, the CD41+ platelets were visualized using an NIR imager. In L-PRF and CGF, platelets were distributed mainly on and below the distal surface, while in bio-PRF and A-PRF, platelet distribution was widespread and homogenous. Among three regions of the PRF matrices (upper, middle, and lower), no significant differences were observed. These findings suggest that platelets aggregate on polymerizing fibrin fibers and float up as a PRF matrix into the plasma fraction, amending the current "gradient" theory of platelet distribution.

Community analysis of biofilms on flame-oxidized stainless steel anodes in microbial fuel cells fed with different substrates.

BACKGROUND: The flame-oxidized stainless steel anode (FO-SSA) is a newly developed electrode that enhances microbial fuel cell (MFC) power generation; however, substrate preference and community structure of the biofilm developed on FO-SSA have not been well characterized. Herein, we investigated the community on FO-SSA using high-throughput sequencing of the 16S rRNA gene fragment in acetate-, starch-, glucose-, and livestock wastewater-fed MFCs. Furthermore, to analyze the effect of the anode material, the acetate-fed community formed on a common carbon-based electrode-carbon-cloth anode (CCA)-was examined for comparison. RESULTS: Substrate type influenced the power output of MFCs using FO-SSA; the highest electricity was generated using acetate as a substrate, followed by peptone, starch and glucose, and wastewater. Intensity of power generation using FO-SSA was related to the abundance of exoelectrogenic genera, namely Geobacter and Desulfuromonas, of the phylum Proteobacteria, which were detected at a higher frequency in acetate-fed communities than in communities fed with other substrates. Lactic acid bacteria (LAB)-Enterococcus and Carnobacterium-were predominant in starch- and glucose-fed communities, respectively. In the wastewater-fed community, members of phylum Planctomycetes were frequently detected (36.2%). Exoelectrogenic genera Geobacter and Desulfuromonas were also detected in glucose-, starch-, and wastewater-fed communities on FO-SSA, but with low frequency (0-3.2%); the lactate produced by Carnobacterium and Enterococcus in glucose- and starch-fed communities might affect exoelectrogenic bacterial growth, resulting in low power output by MFCs fed with these substrates. Furthermore, in the acetate-fed community on FO-SSA, Desulfuromonas was abundant (15.4%) and Geobacter had a minor proportion (0.7%), while in that on CCA, both Geobacter and Desulfuromonas were observed at similar frequencies (6.0-9.8%), indicating that anode material affects exoelectrogenic genus enrichment in anodic biofilm. CONCLUSIONS: Anodic community structure was dependent on both substrate and anode material. Although Desulfuromonas spp. are marine microorganisms, they were abundant in the acetate-fed community on FO-SSA, implying the presence of novel non-halophilic and exoelectrogenic species in this genus. Power generation using FO-SSA was positively related to the frequency of exoelectrogenic genera in the anodic community. Predominant LAB in saccharide-fed anodic biofilm caused low abundance of exoelectrogenic genera and consequent low power generation.

Tooth Germ-Like Construct Transplantation for Whole-Tooth Regeneration: An In Vivo Study in the Miniature Pig.

The purpose of this study was to demonstrate the feasibility of whole-tooth regeneration using a tooth germ-like construct. Dental pulp from upper incisors, canines, premolars, and molars were extracted from sexually mature miniature pigs. Pulp tissues were cultured and expanded in vitro to obtain dental pulp stem cells (DPSCs), and cells were differentiated into odontoblasts and osteoblasts. Epithelial cells were isolated from gingival epithelium. The epithelial cells, odontoblasts, and osteoblasts were seeded onto the surface, upper, and lower layers, respectively, of a bioactive scaffold. The lower first and second molar tooth germs were removed bilaterally and the layered cell/scaffold constructs were transplanted to the mandibular alveolar socket of a pig. At 13.5 months postimplantation, seven of eight pigs developed two teeth with crown, root, and pulp structures. Enamel-like tissues, dentin, cementum, odontoblasts, and periodontal tissues were found upon histological inspection. The regenerated tooth expressed dentin matrix protein-1 and osteopontin. All pigs had regenerated molar teeth regardless of the original tooth used to procure the DPSCs. Pigs that had tooth germs removed or who received empty scaffolds did not develop teeth. Although periodontal ligaments were generated, ankylosis was found in some animals. This study revealed that implantation of a tooth germ-like structure generated a complete tooth with a high success rate. The implant location may influence the morphology of the regenerated tooth.

Use of microscale heterogeneity in samples for spectral factorization-A strategy to build robust prediction models for nondestructive analyses.

Nondestructive spectroscopic analysis is widely used to evaluate food composition. However, distinguishing analytes of interest from other compounds remains challenging. Since most foods are heterogeneous when viewed under a microscope, we hypothesized that spectra measured at microscopic points would be "purer" than spectra acquired from a larger area. By coupling this data with nonnegative matrix factorization (NMF), the analytes of interest can be separated. This preliminary study discusses the quantification of glucose in mixtures of different sugars. Samples were made by mixing glucose with other powders in different ratios and Raman spectra were measured at 200 micro-points for each sample. NMF was applied to factorize the mixed spectra into spectra of pure compounds and their concentrations, leading to the accurate quantification of glucose, while eliminating the effects of other compounds. While this study targets simple powders, separation of analytes using microscale heterogeneity is applicable for measuring more complex foods.

Utilization of multiple-dilution fluorescence fingerprint facilitates prediction of chemical attributes in spice extracts.

Fluorescence Fingerprint (FF) is a powerful tool for rapid quality assessment of various foods and plant-derived products. However, the conventional utilization of FFs measured at a single dilution level (DL) to substitute chemical analyses is extremely challenging, especially for multicomponent materials like spice extracts because fluorescence intensity and concentration widely differ between components, with complex phenomena like inner filter effects. Here, we proposed a new strategy to use the meta-data comprised of FFs measured at multiple DLs with machine learning to estimate common chemical attributes including total polyphenol and flavonoid contents, and antioxidant abilities. This strategy achieved more consistently satisfactory performance in estimation of all chemical attributes of spice extracts compared to using a single DL. Hence, the workflow employed in this study is expected to serve as an alternative method to quickly evaluate the chemical quality of spice extracts, as well as other plant products and food materials.

Sclerosing Odontogenic Carcinoma With a Prominent Clear Cell Component Mimicking Odontogenic Clear Cell Carcinoma: An Extremely Rare Case With a Fatal Clinical Outcome.

Sclerosing odontogenic carcinoma (SOC) is an exceedingly rare odontogenic carcinoma known for its locally aggressive yet indolent behavior. There have been no reports of metastasis to distant organs, except a single case involving lymph node metastasis. This report details the case of a 49-year-old female who presented with a well-demarcated radiolucent lesion in the mandible, accompanied by root resorption and tooth displacement. Microscopically, the lesion exhibited a distinctive composition, with two distinct components: cords of epithelium embedded within an abundant collagenous stroma and solid nests of clear polygonal cells surrounded by hyalinized stroma. Notably, the tumor exhibited direct invasion into the submental muscles, accompanied by perineural and vascular invasion, as well as cortical bone loss. Additionally, the clear cells contained diastase-sensitive periodic acid-Schiff-positive granules. Immunohistochemically, the tumor cells displayed positivity for cytokeratin 19 and p63 while testing negative for myoepithelial markers. The Ki-67 index was measured at 23%. Importantly, neitherEWSR1 nor MAML2 rearrangements were detected through fluorescence in situ hybridization (FISH) analysis. Over several years, this patient experienced three instances of local recurrence; notably, four years after the initial surgery, fludeoxyglucose F18-positron emission tomography (18FDG-PET)/CT scans confirmed the presence of pulmonary metastasis. This case presents an unusual histological variation of SOC, marked by vascular invasion, and is notably the first documented case of a fatal outcome in this context.

Isolation of Yeast and LAB from Dry Coffee Pulp and Monitoring of Organic Acids in Inoculated Green Beans.

Yeast and lactic acid bacteria (LAB) are known to play an important role in the fermentation process of coffee post-harvest. This study aimed to isolate and screen yeast and LAB to be applied in lab-scale refermentation of commercial green coffee beans and coffee pulp with the aim of modifying the composition of organic acids (OAs) in coffee beans. Yeast and LAB strains were isolated from green coffee beans and dry coffee pulp and identified, and their effect on OA concentration in the coffee beans was quantified. In addition, the effects of different fermentation conditions (additional carbon source, different inoculum dose, and different types of coffee pulp) were evaluated based on OA quantification. Nine yeast isolates of Rhodotorula mucilaginosa and Wickerhamomyces anomalus were identified, and 11 LAB isolates of the species Enterococcus mundtii were identified. Of the 7 OAs quantified, quinic acid was the most abundant. The inoculation of isolated yeasts and LAB led to higher concentrations of OAs, showing the potential to realize modification of the OA composition of green coffee beans by re-fermentation with coffee-originated isolates.

Effect of Slightly Acidic Electrolyzed Water Immersion at Different Frequencies on Quality of Raw Chicken Legs.

Slightly acidic electrolyzed water (SAEW) is used as a disinfectant for raw chicken meat. Because its volume for a single immersion exceeds 10 times the weight of meat, a large amount of wastewater is generated. Importantly, a higher frequency of immersion is believed to reduce microbial contamination. The objective of this study was to investigate the effect of SAEW immersion at different frequencies on the disinfection and quality of raw chicken legs, thereby possibly limiting the usage of SAEW. Immersion for 1, 3, and 5 times, with a 7:1 SAEW:meat ratio, and duration of 15 min was tested. Meat quality was evaluated based on total aerobic bacteria, Enterobactericeae, total volatile basic nitrogen, thiobarbituric acid reactive substances, and color. A higher immersion frequency lowered the numbers of total aerobic bacteria and Enterobacteriaceae. Moreover, two immersions with a SAEW:meat ratio of 4:1 and a total immersion time of 6 min reduced the bacterial load as effectively as a single 15-min immersion with a SAEW:meat ratio of 7:1. Higher frequencies of SAEW immersion also resulted in lower total volatile basic nitrogen and lipid oxidation after 0 or 3 days of storage. They did, however, magnify the change in color, resulting in brighter meat. Overall, SAEW treatments with two to five immersions can improve the quality of raw chicken legs and reduce wastewater generation.

Metformin-suppressed platelet's function in vitro: Possible relation to delayed or failure of platelet-rich fibrin preparation.

Platelet-rich fibrin (PRF) is a popular autologous blood-derived biomaterial that is used in regenerative therapy. Owing to its simple preparation without additional factors, the PRF quality directly reflects the characteristics of individual blood samples. Antiplatelet or anticoagulant drugs can hamper the successful preparation of PRF. We recently observed similar phenomena in metformin-taking type-2 diabetics (T2DM). Thus, we hypothesized that metformin interferes with platelet function, thereby suppressing coagulation. For practical reasons, leukocyte- and platelet-rich plasma was prepared from healthy male donors (n = 9-15, age: 26-80 years) and treated with metformin (1-10 mM) for 24-72 h. Intrinsic and extrinsic coagulation activities were evaluated using prothrombin time (PT) and activated partial thromboplastin time (ATPP). Platelet adhesion and aggregation assays were performed using ADP stimulation. Among the parameters tested, APTT was the most sensitive and was significantly prolonged in the concentration range of 1-10 mM in a time- and concentration-dependent manner. Although obtained from healthy platelets and relatively higher concentrations of metformin, these findings suggest that metformin may induce further dysfunction of platelets to suppress intrinsic coagulation activity in T2DM patients, leading to failure of PRF preparation. This phenomenon may not have a severe impact on clinical diabetology or hematology. However, clinicians using PRF are recommended to be more sensitive to such information to avoid unexpected events in clinical settings.

Plasma Gel Made of Platelet-Poor Plasma: In Vitro Verification as a Carrier of Polyphosphate.

Plasma gel (PG) is a blood-derived biomaterial that can be prepared by heating or chemical cross-linking without the aid of intrinsic coagulation activity and has gradually been applied in the field of esthetic surgery. To explore the applicability of PG in regenerative therapy or tissue engineering, in this study, we focused on the advantages of the heating method and verified the retention capacity of the resulting PG for polyphosphate (polyP), a polyanion that contributes to hemostasis and bone regeneration. Pooled platelet-poor plasma (PPP) was prepared from four healthy male adult donors, mixed with synthetic polyP, and heated at 75 °C for 10 or 30 min to prepare PG in microtubes. The PG was incubated in PBS at 37 °C, and polyP levels in the extra-matrix PBS were determined by the fluorometric method every 24 h. The microstructure of PG was examined using scanning electron microscopy. In the small PG matrices, almost all of the added polyP (~100%) was released within the initial 24 h. In contrast, in the large PG matrices, approximately 50% of the polyP was released within the initial 24 h and thereafter gradually released over time. Owing to its simple chemical structure, linear polyP cannot be theoretically retained in the gel matrices used in this study. However, these findings suggest that thermally prepared PG matrices can be applied as carriers of polyP in tissue engineering and regenerative medicine.

Parametric analysis of a novel cryogenic CO2 capture system based on Stirling coolers.

CO(2) capture and storage (CCS) is an important alternative to control greenhouse gas (GHG) effects. In previous work, a novel desublimation CO(2) capture process has been exploited making use of three free piston Stirling coolers (namely, SC-1, SC-2, and SC-3, respectively). Based on the developed system, moisture and CO(2) in the flue gas can condense and desublimate in the prefreezing and main-freezing towers, respectively. Meanwhile, the storage column is chilled by SC-3 to preserve the frosted CO(2), and permanent gas (such as N(2)) passes through the system without phase change. The whole process can be implemented at atmospheric pressure and reduce the energy penalty (e.g., solvent regeneration and pressure drop) in other technologies. In this work, the influence of process parameters has been investigated in detail. The optimal conditions for the system are as follows: idle operating time is 240 min, flow rate is 5 L/min, vacuum degree of the interlayer is 2.2 × 10(3) Pa, and temperatures of SC-1, -2, and -3 are -30, -120, and -120 °C, respectively. Under these conditions, the energy consumption of the system is around 0.5 MJ(electrical)/kg CO(2) with above 90% CO(2) recovery.

Non-destructive, spectrophotometric analysis of the thickness of the cell-multilayered periosteal sheet.

BACKGROUND: Autologous tissue-engineered periosteal sheets, which have been clinically applied for periodontal regeneration, sinus lift, and alveolar ridge augmentation, are enriched with osteoblast precursor cells and the abundant deposition of collagen type I in the extracellular spaces. Their quality is inspected prior to clinical use; however, most criteria cannot be evaluated without sacrificing samples. To reduce such losses, we developed a non-destructive optical method that can quantitatively evaluate the thickness of the periosteal sheet. METHODS: Dispersed periosteal cells were inoculated into small pieces of collagen sponge (Terudermis®) and plated into 60-mm dishes for further explant culture using a conventional medium and a stem-cell culture medium. The thickness of periosteal sheets was evaluated using inverted microscopic, histological, labeling (CellVue®)-based imaging and spectrophotometric (Spectro-1®) methods. RESULTS: The three-dimensional growth of periosteal sheets did not necessarily correlate with two-dimensional growth. The periosteal sheet prepared with the stem-cell medium formed cell multilayers, a phenomenon that could be observed qualitatively by inverted microscopy. The spectrophotometric analysis enabled the quantitative evaluation of the thickness of the cell multilayer without sacrificing the samples processed for scheduled cell therapy. CONCLUSIONS: The growth of periosteal sheets is influenced by several major factors, including the basic quality of the individual original periosteal tissue segments, the technical expertise of doctors and operators involved in tissue harvesting and processing, and culture conditions. This newly developed spectrophotometric analysis can quantify the thickness of cell-multilayered periosteal sheets for quality assurance in a non-destructive manner, thereby contributing to better bone augmentation prior to implant therapy.

Effects of SARS-CoV-2 mRNA vaccines on platelet polyphosphate levels and inflammation: A pilot study.

Platelets function as immune cells in conjunction with white blood cells, targeting invading pathogens and inducing immune reactions. Intercellular communications among these immune cells are partly mediated by platelet polyphosphate (polyP), which was originally recognized as a thrombotic and hemostatic biomolecule. To determine the involvement of polyP in SARS-CoV-2-mRNA vaccine-induced immune responses, specifically in inflammatory responses, the effects of mRNA vaccines on platelet polyP levels were examined. Before and after vaccination with the COVID-19 vaccine (BNT162b2), blood samples were obtained from healthy, non-smoking individuals who did not have any systemic diseases. Test group demographics skewed somewhat towards either older males (first vaccination, n=6; second vaccination, n=8) or younger females (first vaccination, n=14; second vaccination, n=23). polyP levels in washed platelets from the blood samples were determined using the fluorometric method with DAPI. The side-effects of vaccination were recorded as scores. In the female group, platelet polyP levels decreased after the first vaccination, and the side-effect score increased after the second vaccination. Moderate correlation coefficients were observed between the reduction in polyP levels and the side-effect scores and pre-vaccination polyP levels. Despite the small sample size, this pilot study suggests that platelet polyP may suppress the side effects induced by the mRNA vaccines after the first vaccination, but not the second vaccination in younger female subjects, who generally have higher immune responsiveness than their male counterparts.

Part II: crystalline fluorapatite-coated hydroxyapatite implant material: a dog study with histologic comparison of osteogenesis seen with FA-coated HA grafting material versus HA controls: potential bacteriostatic effect of fluoridated HA.

Success of osteogenesis in bone graft procedures can be enhanced by inhibiting oral bacterial infections through the use of prophylactic bacteriostatic fluoride within the grafting environment. Ideally, the fluoride ion should be chemically sequestered and thus unavailable unless needed at times during the process of early infection. As fluoride within fluorapatite is tightly bound at neutral pH and becomes available only during acidic conditions, fluorapatite is an ideal store for the fluoride ion which becomes released for bacteriostasis only during an acidic environment found with incipient bacterial infection. The purpose of this investigation was to compare the histologic properties of new bone formed surrounding fluorapatite (FA)-coated microcrystalline hydroxyapatite (HA) grafting material with comparable bone formed following the use of control HA material (OsteoGen, Impladent, Ltd, Holliswood, NY). The results of histologic analysis within dog studies here showed no detectable difference in new bone following therapeutic grafting procedures using each of the above 2 mineral coatings.

First Clinical Trial of a Newly Developed, Low Menaquinone-7, Fermented Soybean Natto in Warfarin-Dependent Patients.

Bacillus subtilis fermented soybeans (natto) contain high vitamin K2 levels, mostly as menaquinone-7 (MK-7), and must be avoided by warfarin-dependent patients. This is the first report which demonstrates the characteristics and clinical relevance of a low MK-7 natto for such patients. We generated a novel, mutant B. subtilis strain TTCC2051 with short-term fermentation and reduced MK-7 production, yielding 19-24% of the normal MK-7 content. After functional assessments and a preclinical trial, 10 warfarin-dependent patients underwent a clinical trial with a 7-day ingestion test of the low MK-7 natto. Functional assessments were satisfactory, and the preclinical trial showed no increases in plasma MK-7 levels after 7 days of ingestion. In the clinical trial, 20 g/day of the low MK-7 natto significantly increased plasma MK-7 levels while 10 g/day did not. However, neither dose of low MK-7 natto changed international normalized ratio of prothrombin time (PT-INR) values in either group. The low MK-7 natto neither changed PT-INR values nor precipitated adverse events if ingested with a once-daily maximum of 20 g (46 µg of MK-7). Thus, this novel food product has potential for consumption by warfarin-dependent patients.