Advances of nanopore-based sensing techniques for contaminants evaluation of food and agricultural products.

Food safety assurance systems are becoming more stringent in response to the growing food safety problems. Rapid, sensitive, and reliable detection technology is a prerequisite for the establishment of food safety assurance systems. Nanopore technology has been taken as one of the emerging technology capable of dealing with the detection of harmful contaminants as efficiently as possible due to the advantage of label-free, high-throughput, amplification-free, and rapid detection features. Start with the history of nanopore techniques, this review introduced the underlying knowledge of detection mechanism of nanopore-based sensing techniques. Meanwhile, sensing interfaces for the construction of nanopore sensors are comprehensively summarized. Moreover, this review covers the current advances of nanopore techniques in the application of food safety screening. Currently, the establishment of nanopore sensing devices is mainly based on the blocking current phenomenon. Sensing interfaces including biological nanopores, solid-state nanopores, DNA origami, and de novo designed nanopores can be used in the manufacture of sensing devices. Food harmful substances, including heavy metals, veterinary drugs, pesticide residues, food toxins, and other harmful substances can be quickly determined by nanopore-based sensors. Moreover, the combination of nanopore techniques with advanced materials has become one of the most effective methods to improve sensing properties.

Advancements of nature nanocage protein: preparation, identification and multiple applications of ferritins.

Ferritin is an important iron storage protein, which is widely existed in all forms of life. Ferritin can regulate iron homeostasis when iron ions are lacking or enriched in the body, so as to avoid iron deficiency diseases and iron poisoning. Ferritin presents a hollow nanocage, which can store ions or other small molecular substances in the cavity. Therefore, ferritin shows its potential as a functional nanomaterial that can deliver nutrients or drugs in a targeted manner to improve bioavailability. Due to the special structure, the research on ferritin has attracted more and more attention in recent years. In this paper, the structural characteristics of ferritin were introduced, and the natural purification and prokaryotic expression methods of ferritin from different sources were described. At the same time, ferritin can bind to small molecules, so that it has the activity of small molecules, to construct a new type of ferritin. As a result, ferritin plays an important role as a nutrient substance, in targeted transport, and disease monitoring, etc. In conclusion, the yield of ferritin can be improved by means of molecular biology. Meanwhile, molecular modification can be used to make ferritin have unique activity and function, which lays a foundation for subsequent research.HighlightsThe molecular and structural properties of ferritins were clearly described.Isolation and purification technologies of ferritin were compared.Characterization, functions and molecular modifications mechanism of ferritin were reviewed.The applications of ferritin in pharmaceutical and food industry were prospected.

Over 20 years of chronic patellar ligament rupture with severe knee osteoarthritis for Total knee Arthroplasty: a case report.

BACKGROUND: Extensor apparatus rupture is a severe complication after knee arthroplasty, but there have not been many reports on how to perform knee arthroplasty after chronic patellar ligament rupture. We reported a case of total knee arthroplasty (TKA) in a patient with severe osteoarthritis (OA) complicated by chronic patellar ligament rupture. CASE PRESENTATION: In this case, a 67-year-old male patient suffered from patellar ligament rupture due to trauma more than 20 years ago and did not undergo any formal treatment. Physical examination revealed a small amount of fluid and extension lag, and the patella was displaced upward by approximately 5.5 cm. The quadriceps were atrophic and weak. There was significant tenderness on the medial side of the left knee joint. Passive motion of the left knee joint ranged from full extension to 120° of flexion with discomfort during excessive flexion. Active flexion of the knee joint to 120°, and extensor lag was approximately 90°. We reconstructed the extensor apparatus through a quadriceps tendon V-Y quadricepsplasty and Krackow suture technique of the patellar ligament, and osteoarthritis was resolved with TKA. The visual analogue scale (VAS) score decreased from 5 points to 1 point after surgery. Six weeks later, the patient was able to walk normally without a walking stick, and the knee joint could stretch actively to approximately 30°. However, he had obvious extension lag. This problem improved 10 months after surgery. The AKS score increased from 35 to 95 10 months after surgery. The HSS score increased from 43 to 93. CONCLUSIONS: TKA and ligament reconstruction are options for the treatment of knee OA with chronic patellar ligament rupture. V-Y lengthening of the quadriceps femoris tendon after the Krackow suture technique of the patellar ligament with transpatellar tunnels may be a reasonable choice during TKA.

Reliability and reproducibility analysis of the Cobb angle and assessing sagittal plane by computer-assisted and manual measurement tools.

BACKGROUND: Although many studies on reliability and reproducibility of measurement have been performed on coronal Cobb angle, few results about reliability and reproducibility are reported on sagittal alignment measurement including the pelvis. We usually use SurgimapSpine software to measure the Cobb angle in our studies; however, there are no reports till date on its reliability and reproducible measurements. METHODS: Sixty-eight standard standing posteroanterior whole-spine radiographs were reviewed. Three examiners carried out the measurements independently under the settings of manual measurement on X-ray radiographies and SurgimapSpine software on the computer. Parameters measured included pelvic incidence, sacral slope, pelvic tilt, Lumbar lordosis (LL), thoracic kyphosis, and coronal Cobb angle. SPSS 16.0 software was used for statistical analyses. The means, standard deviations, intraclass and interclass correlation coefficient (ICC), and 95% confidence intervals (CI) were calculated. RESULTS: There was no notable difference between the two tools (P = 0.21) for the coronal Cobb angle. In the sagittal plane parameters, the ICC of intraobserver reliability for the manual measures varied from 0.65 (T2-T5 angle) to 0.95 (LL angle). Further, for SurgimapSpine tool, the ICC ranged from 0.75 to 0.98. No significant difference in intraobserver reliability was found between the two measurements (P > 0.05). As for the interobserver reliability, measurements with SurgimapSpine tool had better ICC (0.71 to 0.98 vs 0.59 to 0.96) and Pearson's coefficient (0.76 to 0.99 vs 0.60 to 0.97). The reliability of SurgimapSpine measures was significantly higher in all parameters except for the coronal Cobb angle where the difference was not significant (P > 0.05). CONCLUSION: Although the differences between the two methods are very small, the results of this study indicate that the SurgimapSpine measurement is an equivalent measuring tool to the traditional manual in coronal Cobb angle, but is advantageous in spino-pelvic measurement in T2-T5, PT, PI, SS, and LL.

Properties regulation and mechanism on ferritin/chitooligosaccharide dual-compartmental emulsions and its application for co-encapsulation of curcumin and quercetin bioactive compounds.

Dual-compartmental emulsions, containing multiple chambers, possess great advantages in co-encapsulation of different cargoes. Herein, we reported a stable dual-compartmental emulsion by regulating the ratio of Marsupenaeus japonicus ferritin (MF) and chitooligosaccharide (COS), enabling efficient co-encapsulation of different compounds. The adsorption behavior of MF/COS complex over droplet interface varied at different ratios, thereby exerting an influence on the emulsion properties. Remarkably, emulsions stabilized by MF/COS complex at a ratio of 2:1 exhibited superior stability, as evidenced by no significant creaming or demulsification during storage or heat treatment. The mechanism is that MF/COS2:1 complex can enhance the formation of thicker interfacial layer and dense continuous phase network structure. Additionally, curcumin and quercetin can be co-encapsulated into the emulsions and their retention rates were significantly improved than those in oils, implying the potential of the resulting dual-compartmental emulsions in co-encapsulation and delivery of bioactive compounds.

[Arthroscopic surgery for injuries to the popliteal tendon area of the lateral meniscus].

OBJECTIVE: To investigate the mid-term effect and complications of arthroscopic popliteal tendon suture in the treatment of lateral meniscus injury. METHODS: From January 2016 to December 2020, the data of 57 patients with lateral meniscus popliteal tendon injury treated by arthroscopic popliteal tendon suture fixation were retrospectively analyzed, including 35 males and 22 females, aged from 18 to 47 years old with an average of (32.9±7.9) years old. Knee function was evaluated using the International Knee Documentation Committee (IKDC) and Lysholm scores both before the operation and at the final follow-up. Meniscus healing was evaluated according to the postoperative Barrett standard. Wound healing complications, such as vascular injury, nerve injury, and lower extremity venous thrombosis, were recorded. RESULTS: All 57 patients were followed up for 12 to 58 months with an average of (38.1±14.9) months.The incisions of the patients after the operation were all Grade A healing without infection, popliteal tendon injury, blood vessel injury, nerve injury and lower extremity venous thrombosis.The IKDC score increased from (49.7±3.6) points preoperatively to (88.5±4.4) points in the final follow-up (P<0.05). The Lysholm score increased from (48.8±4.9) points preoperatively to (91.9±3.9) points at the final follow-up (P<0.05). At 3, 6 months and 1 year after operation, according to Barrett's criteria, 54 cases were clinically healed, the healing rate was 94.7% (54/57). CONCLUSION: This study preliminarily confirmed that arthroscopic suture technique can result in clinical stability through suture and fixation of the meniscus in the injured lateral popliteal tendon area. No adverse effects on knee joint function were found in the mid-term follow-up after the operation.

Genipin-mediated subunit-subunit crosslinking of ferritin nanocages: Structure, properties, and its application for food bioactive compound sealing.

Ferritin proteins are promising nano-carriers for bioactive compound delivery. However, the disassembly properties under acidic/alkaline conditions pose risks of cargo leakage. Herein, genipin-mediated chemical crosslinking method was provided as an alternative and effective strategy to construct robust ferritin nanocarrier through controlled-intramolecular conjugation. As indicated by SDS-/Native- PAGE, the crosslinking degree gradually increased with incubating time prolonging. CD results showed that the cross-linking would decrease α-helix content from 78.4 % to 52.7 % upon 6 h incubation. However, TEM images showed that the genipin-modification has subtle influence on its shell-like structure. Remarkably, the cross-linking can be well controlled by intramolecular subunit-subunit conjugation rather than intermolecular conjugation, giving an excellent monodispersity. Importantly, the covalent cross-linking can tight neighboring subunits and inhibit its disassociation, finally inhibiting the leakage of encapsulated-cargos from ferritin cavity under acidic environments. Such findings suggested that the genipin-mediated cross-linking strategy can fabricate robust nano-carriers for bioactive compound delivery.

PM1-loaded recombinant human H-ferritin nanocages: A novel pH-responsive sensing platform for the identification of cancer cells.

The aggregation-induced emission (AIE) material has been widely used in biological detection due to their unique property of fluorescing in aggregation state. However, the poor dispersion and biocompatibility limit its application in in vivo real-time imaging. Here, a novel strategy is designed to obtain pH-responsive AIE nanomaterials, working through 4-Undecoxy Tetraphenyl Ethylene Methacrylate (PM1) block, with excellent features (dispersion, biocompatibility, self-reconstruction and cancer specific recognition). The recombinant human H-ferritin (rHuHF) was used to prepare rHuHF-PM1 nanocomposites which effectively supported the dispersion and transfer of PM1 in the biological environment, even making it target tumor cells due to the overexpression of ferritin receptors on tumor cells. To simulate the changes of rHuHF in intracellular lysosomes, particle size and fluorescence of rHuHF-PM1 were analyzed, which reflected the loose structural changes of rHuHF nanocages in weak acid system that facilitated the degradation of macromolecular rHuHF in intracellular lysosomes and following release of PM1. The released PM1 molecules aggregated and emitted brilliant blue fluorescence. Several cell lines, Hela, HT-29, HepG2, L-O2 and HUVEC have all been sensitively detected and distinguished. Accordingly, this nanocage has a potential to be applied to disease diagnosis and provides a novel sensing platform for the identification of cancer.

Ferritin nanocage based delivery vehicles: From single-, co- to compartmentalized- encapsulation of bioactive or nutraceutical compounds.

Bioactive or nutraceutical ingredients have been widely used in pursuit of health and well-being. However, the environmental instability, poor solubility and bioavailability, and unspecific delivery highly limited their practical values. By virtue of the unique shell-like structure, definite disassembly/reassembly behavior, and excellent safety profile of ferritin protein, it stands out among of various nano-materials and is emerging as one of the most promising vehicles for the encapsulation and delivery of bioactive ingredients or drugs. In this review, we present a systematic overview of recent advances of ferritin-based delivery systems from single-encapsulation, co-encapsulation, to compartmentalized-encapsulation of bioactive ingredients or drugs. Different encapsulation strategies for cargo loading as well as their advantages and drawbacks have been critically reviewed. This study emphasized the importance of the construction of compartmentalized delivery systems through the usage of ferritin nanocages, which exhibit great potential for facilitating the synergistic functionality of different types of cargos. Lastly, the applications of ferritin nanocages for physicochemical improvements and functionality achievements of loaded cargos are summarized. In conclusion, ferritin protein nanocages not only are excellent nanocarriers, but also can act as"multi-seated" vehicles for co-encapsulation and compartmentalized encapsulation of different cargos simultaneously.

Oyster (Crassostrea gigas) ferritin can efficiently reduce the damage of Pb2+in vivo by electrostatic attraction.

Heavy metal ions pollution can cause damage to human body through food, so the development of a new kind of macromolecular that can remove heavy metal ions damage has a good application prospect. The possibilities of removing heavy metal ions from food system with ferritin were studied in this paper. In this study, oyster ferritin (GF1) can resistant to denaturation induced by Pb2+, Cd2+, Cr3+ and still maintains its basic structure. GF1 can bind more Pb2+, Cd2+, Cr3+ than recombinant human H-chain ferritin (rHuHF), especially Pb2+, and the findings suggest that each GF1 can capture about 51.42 Pb2+ in solution. The hard and soft acids and base also verifies that Pb2+ have stronger binding ability to the key amino acids at the outer end of the three-fold symmetry channel. Cells preprotected by ferritin could resistant to heavy metal ions. And GF1 can reduce the high blood lead in mice and may play a role in alleviating lead poisoning in vivo. All findings demonstrated that GF1 can be used as a novel macromolecule to bind heavy metal ions, and the study can broaden the research scope of ferritin in contaminated food systems.

Shrimp ferritin greatly improves the physical and chemical stability of astaxanthin.

The low stability of trans-astaxanthin (AX) not only limits its applications as a functional factor in food systems, but also affects the sensor quality of most shrimp products. Therefore, it is important to find an easy, effective way to improve the physical and chemical stability of AX. In this study, by taking advantage of the co-existence of AX and shrimp ferritin (Marsupenaeus japonicus ferritin, MjF), we investigated the interaction of AX with MjF. Results showed that AX molecules are able to bind on the outer surface of MjF to form complexes, and quantitative analyses demonstrated that one ferritin molecule is bound to ∼48 AX molecules. Consequently, such binding not only greatly enhances the water solubility, thermal stability, and photo stability of AX, but also protects AX from Fe2+ -induced oxidative damage, as compared to free AX. Thus, MjF could be used as a protective molecule to improve the physical and chemical stability of AX. PRACTICAL APPLICATION: Our study opens up a new avenue for improving the physicochemical properties of bioactive molecules by interacting with protein, and shrimp ferritin could be explored as a protective system for the bioactive molecules.

Hyperthermostability of prawn ferritin nanocage facilitates its application as a robust nanovehicle for nutraceuticals.

The favorable physicochemical properties are essential for the application of protein-based nanovehicles in the field of biomaterials. Herein, we found that the thermal stability of Marsupenaeus japonicus ferritin (MjFer) (Tm = 109.1 ± 0.4 °C) is markedly higher than human H-chain ferritin (HuHF) (Tm = 87.7 ± 0.3 °C), although they share a high structural similarity. Multiple results indicated that the promoted thermal stability of MjFer is mainly derived from the salt bridges located at the C3 interface. Consequently, MjFer exhibits strong protective effects on encapsulated curcumin upon exposure at high temperatures. In contrast, most of the curcumin loaded HuHF composites precipitated rapidly under the same conditions. These findings elucidated the molecular mechanism of the hyperthermostability of MjFer and illustrated that MjFer could act as a robust insulation nanocarrier for bioactive compounds against various thermal treatments.

Improvement of thermal stability of oyster (Crassostrea gigas) ferritin by point mutation.

Ferritin can be widely used as functional nanomaterial. But the physiological activity of ferritin can be damaged under excessive temperatures, which affect the self-assembly property. In this study, point mutation was produced in Asp120 to Gly120 of ferritin amino acid sequence and the heat resistance was improved significantly. The thermal denaturation temperature of mutated ferritin is 89.17 °C and has increased by 13 °C more than the wild-type oyster ferritin. The effect of thermal treatment on the denaturation, aggregation state, particle size and the structure of ferritin was not changed before 90 °C. The computational modeling and analysis indicated that mutated ferritin promotes the overall structural stability assembly via decreasing the interaction energies of 62 percent energies in 3-fold interface. Improving the thermal stability of oyster ferritin by point mutation enhances its applications as a food ingredient.

Construction of thermally robust and porous shrimp ferritin crystalline for molecular encapsulation through intermolecular arginine-arginine attractions.

Protein colloid crystals are considered as high porous soft materials, presenting great potentials in nutrients and drug encapsulation, but protein crystal fabrication usually needs precipitant and high protein concentration. Herein, an easy implemented approach was reported for the construction of protein colloid crystals in diluted solution with shimp ferritin as building blocks by taking advantage of the strength of multiple intermolecular arginine-arginine interactions. The X-ray single-crystal structure reveals that a group of exquisite arginine-arginine interactions between two neighboring ferritin enable them self-assembly into long-range ordered protein soft materials. The arginine-arginine interactions mediate crystal generation favored at pH 9.5 with 200 mM NaCl, and the resulting colloid crystals exhibit high thermal stability (90 °C for 30 min). Importantly, the interglobular cavity in colloid crystals is three times larger in volume than that of intrinsic ferritin cavity in each unit cell, which can be used for molecular encapsulation.

Converting histidine-induced 3D protein arrays in crystals into their 3D analogues in solution by metal coordination cross-linking.

Histidine (His) residues represent versatile motifs for designing protein-protein interactions because the protonation state of the imidazole group of His is the only moiety in protein to be significantly pH dependent under physiological conditions. Here we show that, by the designed His motifs nearby the C4 axes, ferritin nanocages arrange in crystals with a simple cubic stacking pattern. The X-ray crystal structures obtained at pH 4.0, 7.0, and 9.0 in conjunction with thermostability analyses reveal the strength of the π-π interactions between two adjacent protein nanocages can be fine-tuned by pH. By using the crystal structural information as a guide, we constructed 3D protein frameworks in solution by a combination of the relatively weak His-His interaction and Ni2+-participated metal coordination with Glu residues from two adjacent protein nanocages. These findings open up a new way of organizing protein building blocks into 3D protein crystalline frameworks.

Thermal treatment modified the physicochemical properties of recombinant oyster (Crassostrea gigas) ferritin.

The unique nanocage structure of ferritin can be used as functional nanomaterials and has wide application prospects. However, thermal treatment may affect the structure of ferritin, further affecting self-assembly property. In this study, the oyster ferritin gene GF1 was obtained, prokaryotically expressed in E. coli BL21 (DE3). Then the purified ferritin was heated from 60 to 100 °C for 10 min with untreated ferritin as a control sample. The aggregation state of ferritin was investigated and the difference in protein structure was evaluated in terms of particle size and protein structures. The results of electrophoresis indicated that thermal treatment induced denaturation and aggregation of ferritin macromolecules. Moreover, the particle size distribution shifted to larger size trend and aggregates were visible with the heated samples and the secondary structure and tertiary structure were destroyed gradually. The findings are beneficial for understanding the relationship of the structure and function of ferritin.

Relationship between enzyme, peptides, amino acids, ion composition, and bitterness of the hydrolysates of Alaska pollock frame.

Alaska pollock frame is a kind of byproduct that was rich in protein, amino acids, and mineral elements. However, the unfavorite bitterness may probably be produced in enzymatic processes. In this study, the bitterness accounted from the hydrolysates prepared by neutral proteases, alkaline proteases, papain, flavourzyme, and animal proteases, was investigated. The hydrolysis conditions, amino acids composition, metal ion composition, molecular weight distribution, and peptide composition of the hydrolysates were detected to figure out the relationship between bitterness and compositions of the hydrolysates. The hydrolysate digested by alkaline protease has the highest bitterness intensity, and that amino acids composition, peptide composition, and molecular weight distribution had a significant influence on the bitterness degree. Hydrophobic amino acids and alkaline amino acids, such as leucine, isoleucine, lysine, and so on are likely to contribute to the bitterness and molecular weight distribution of peptides that affect bitterness is mainly lower than 3,000 Da. PRACTICAL APPLICATIONS: Fish bones would produce a bitter taste when reusing them by hydrolysis. Bitterness is one of unfavorable flavor as to consumers. The results of this study are of great significance for the further utilization of Alaska pollock frame. For products obtained from the hydrolysate of Alaska pollock frame, such as condiments and health care product the results of this study provide the processing technology of the lowest bitter hydrolysate, which can effectively improve the flavor and acceptability of the products.

[Clinical observation of high tibial osteotomy for knee osteoarthritis:10 years follow-up].

OBJECTIVE: To explore long-term following-up clinical effects of lateral closed high tibial osteotomy for the treatment of knee osteoarthritis. METHODS: Twenty patients with medial unicompartmental knee osteoarthritis were treated with lateral closed high tibial osteotomy and screw fixation from June 2005 to December 2015. Among them, including 17 females and 3 males, aged from 43 to 76 years old with an average of (57.80±8.05) years old. VAS score and KSS score were applied to evaluate recovery degree of pain and function before operation and after operation at 1, 5 and 10 years, and postoperative complications were observed. RESULTS: Sixteen patients were followed-up, the time ranged from 9 to 11(10.0±0.8) years, 4 patients were loss to follow-up. Preoperative VAS score was 7.88±1.15 and decreased to 3.19±0.91, 3.44±0.96, 3.69±1.20 at 1, 5 and 10 years after operation, and there were statistical differences in VAS score between before and after operation at different time points (P<0.05). Clinical score of KSS increased from 61.94±5.74 before opertaion to 75.50±4.62, 80.13±3.97, 77.38±6.40 at 1, 5 and 10 years after operation, and there were statistical differences in clinical score of KSS between before and after operation at different time points(P<0.05); functional score of KSS increased from 62.81±13.03 before operation to 77.50±8.56, 81.88±6.55, 76.88±10.78, and there were statistical differences in functional score of KSS between before and after operation at different time points(P<0.05). All incisions healed well without complications such as fibula nerve injury and fracture nonunion. CONCLUSIONS: Lateral closed high tibial osteotomy and screw fixation for knee osteoarthritis could receive good clinical results, stop and delay progress of knee osteoarthritis, and long-term following-up could achieve the same effect as total knee arthroplasty.

Analysis of volatile compounds and nutritional properties of enzymatic hydrolysate of protein from cod bone.

Hydrolysis of the proteins from cod bone was performed using flavourzyme and trypsin. The nutritional properties of hydrolysates by flavourzyme (HF) and trypsin (HT) were investigated. By comparison, HF exhibits a better degree of hydrolysates (DH) and nitrogen recovery (NR) than HT. The protein content of extract is 97.39% and had a good nutritional value due to the high protein digestibility-corrected amino acid score (PDCAAS) of 0.95 for adults. The content of total amino acids is 942.55 mg/g. The free amino acids content of hydrolysates derived from flavourzyme is 136.82 mg/g after hydrolyzing 3 h, while 17.12 mg/g of such hydrolysates was obtained by using trypsin. The main flavor compounds are alcohols, aldehydes, ketones, acids and alkanes. The molecular weight of predominant peptides ranged from 1000 to 3000 Da in both enzymatic hydrolysates. This study provided a theoretical basis to the preparation of nutritional components with attracting flavor in functional food industry.

Preparation of the rainbow trout bone peptides directed by nutritional properties and flavor analyses.

Rainbow trout bone proteins were prepared by heating at 121°C for 30 min, followed by filtration, concentration, and lyophilization. Nutritional properties and flavor analyses of hydrolysates digested by five different enzymes were investigated, respectively. Results showed that the crude protein content of rainbow trout bone was 15.90% and had a well-balanced nutritional value. The content of total amino acids was 983.64 mg/g. The amount of free amino acids of hydrolysates digested by alkaline protease, neutral protease, flavourzyme, papain, and trypsin for 3 hr was 207.83, 224.13, 1,001.59, 283.26, and 303.64 mg/g, respectively. During the hydrolysis, the main flavor compounds were identified by GC-MS to be alcohols, aldehydes, ketones, acids, and alkanes. After hydrolysis, the main molecular weight of peptides was focused on the range of 1,000-3,000 Da in all enzymatic hydrolysates. This study provided a theoretical basis to comprehensive utilization of rainbow trout bone in food industry.