Half of the test samples (n = 10/group) were fatigued in a mouth-motion chewing-simulator  with simultaneous thermocycling (5–55 °C). All specimens (fatigued and non-fatigued) were afterwards exposed to single-load-to-failure-testing . Statistical analysis was performed using ANOVA, Tukey’s post-hoc tests and two-sample t-tests . All specimens withstood fatigue application. While the effect of fatigue was not significant in any group (p = 0.714), the choice of material had a significant effect (p < 0.001). Material FL recorded the highest failure loads, followed by Z-ST, Z-HT and RL, both with and without fatigue application. Taken together, all tested FDP material combinations survived chewing forces that exceeded physiological levels. Bi-Layer FL and monolithic Z-ST showed the highest resilience and might serve as reliable prosthetic reconstruction concepts for 3-unit FDPs on ceramic implants.No evidence-based prosthetic treatment concept for 3-unit fixed-dental-prostheses (FDPs) on ceramic implants is currently available. Therefore, the aim of this in vitro study was to investigate the failure load and fatigue behavior of monolithic and bi-layer zirconia FDPs supported by one-piece ceramic implants. Eighty 3-unit FDPs supported by 160 zirconia-implants  were divided into 4 groups ( Today, a large variety of dental materials are currently available for reconstructive dentistry and dental implantology. Modern dental ceramics can be subdivided into esthetic glass ceramics, resin-matrix ceramics and high-strength zirconia ceramics ,2. Dental implants consisting of pure titanium or titanium alloys are considered to be the gold-standard in replacing missing teeth, due to their biocompatibility, fatigue strength and good long-term results ,4. HowevLately, ceramic implants have received increasing interest from dentists and patients due to their positive esthetic outcome and tooth-like appearance . Modern A recent systematic review reported a cumulative survival rate of 95% after 5 years for all-ceramic single crowns (SCs) and 3-unit fixed dental prostheses (FDPs) supported by ceramic implants . A detaiAs traditional veneering systems are prone to delamination failures and chip-off fractures, CAD/CAM fabricated monolithic zirconia restorations should be chosen for posterior SCs and FDPs on implants to decrease technical complications ,16. InnoAs with glass ceramics, zirconia restorations benefit from adhesive bonding, especially in cases of compromised retention, high shear forces and minimum ceramic thicknesses ,21. PrerCurrently, two diametrically opposed paths are being pursued to achieve both esthetic and durable, high-strength restorations. On the one hand, multilayer zirconia discs with color and strength gradient technology were launched to merge the positive mechanical and optical properties of different zirconia generations; on the other hand, the combination of different all-ceramic bi-layer materials is used for esthetic restorations ,26. ThisCurrently, no robust prosthetic treatment concept exists for prosthetic reconstructions on ceramic implants. Moreover, wear of zirconia-based restorations is another relevant topic that is extensively discussed within the scientific community, especially when both jaws are restored with different prosthetic materials. So far, no study has investigated possible novel treatment concepts to overcome these issues. A recent in vitro study presented a restorative rapid-layer concept with a polymer-infiltrated ceramic network (PICN) material and a 3Y-TZP framework ,29. ThisAs neither in vitro nor clinical studies have analyzed the performance of monolithic translucent zirconia restorations or the rapid-layer concept of a PICN material with a zirconia framework as FDPs on ceramic implants, this needs to be investigated. Therefore, the aim of this in-vitro study was to assess the thermo-mechanical fatigue behavior and failure modes of monolithic and bi-layer zirconia FDPs supported by one-piece zirconia implants. The tested null hypotheses assumed: There is no difference in (i) failure load and (ii) fatigue behaviour of bi-layer and monolithic implant restorations on zirconia implants. Monolithic 3Y-TZP implant FDPs served as a control. n = 20 per group) depending on the prosthetic material Group Z-ST: monolithic 4Y-TZP zirconia FDP Group FL: 3Y-TZP zirconia FDP  with facial veneer Group RL: polymer-infiltrated ceramic network  “tabletop” resin bonded to a 3Y-TZP zirconia framework In total, 80 FDPs with two one-piece zirconia implants , serving as abutments for a 3-unit FDP, were used. The 80 FDP specimens were split into three test groups and one control group . An optical impression was made  and a master design of a 3-unit FDP  was used for all monolithic FDPs (Group Z-HT and Z-ST) to ensure identical and comparable test samples A. Group 2  and 35 mm2 . The zirconia FDPs were milled in a five-axis milling unit . Polymer-infiltrated ceramic (PICN) Tabletops of Group RL were milled out of industrially prefabricated CAD/CAM blocks  in a three-axis milling machine .For Group RL, a split design from the original master design was used to generate a separate framework and veneer layer C. All im® CW20/ Ren® HY 49, Huntsman Advanced Materials, Salt Lake City, UT, USA) [All zirconia implants were perpendicularly embedded in epoxy resin  ,31. ClinUT, USA) ,32 and 0UT, USA) .The intaglio surface of the zirconia FDPs of all groups were air-particle abraded with 50 μm aluminum-oxide at a pressure of 2 bar prior to adhesive bonding. Prosthetic restorations were subsequently cleaned with 70% ethanol for 3 min in an ultrasound bath , followed by the application of an MDP-containing primer . Polymer-infiltrated ceramic “tabletops” of Group RL were etched with 5% hydrofluoric acid  for 60 s, cleaned with water and dried with oil-free air and then a primer (Clearfil Ceramic Primer Plus) was applied. PICN “tabletops” were then adhesively bonded with a self-curing composite cement  to the air-particle abraded and MDP pretreated occlusal surface of the corresponding 3Y-TZP zirconia framework. The MDP Primer (Clearfil Ceramic Primer) was also applied to the zirconia implants. Afterwards, all zirconia FDPs were bonded with a self-curing composite (Panavia V5 opaque) to the one-piece ceramic implants. To allow a complete polymerization of the adhesive interface, all specimens were put in distilled water at 37 °C in an incubator  for 24 h . Table 2Ten specimens from each group were subjected to dynamic thermomechanical loading  in a mouth motion fatigue simulator . An occlusal load of 98 N at 1.2 million chewing cycles with a frequency of 1.6 Hz was applied to the central fossa of the pontic of the restoration, equivalent to a 5-year clinical service time ,35,36. AAll samples (loaded and non-loaded) were vertically loaded until failure in a universal testing machine . Failure load was applied at the same contact point as during the chewing simulation with a crosshead speed of 1.5 mm/min. A steel ball with a diameter of 6 mm served as a load indenter. Fractures of the ceramic veneer  and catastrophic core fractures of the framework/monolithic FDP restoration were defined as failure.Firstly, test specimens were investigated under a polarized light microscope  after fatigue and SLF. Most meaningful samples were then analyzed for qualitative fractographic evaluation with a scanning electron microscope  to determine the failure mode. n = 10 per group  was found to enable detection of any effect of at least large size (Cohen’s effect size of f = 0.38) with 80% power and a two-sided type-I error of p < 0.05 for the two factors and their interaction. A power calculation  with the factors (i) type of material  and (ii) fatigue application (yes/no) of bi-layer and monolithic FDPs was performed with respect to statistical testing via ANOVA. A sample size of t-tests were used to test separately for influence of fatigue for each type of material. The p-value was defined at <0.05 (95% CI) for all tests, Bonferroni-corrected for multiple comparisons where applicable. Data were graphically displayed in boxplots. The statistical software SPSS 28  was used to perform the analysis. To test for homogeneity of variance before using ANOVA, the Levene Test was applied to test for the main effects and interactions of the two factors of interest , followed by Tukey’s post-hoc tests for the pairwise comparison of material types. Two-sample All investigated samples survived thermomechanical loading, leading to a 100% 5-year simulated survival rate. No bulk or cohesive fractures within the FDPs or implants were noticed during and after chewing simulation. Only superficial wear of the glazing material could be observed for Groups Z-HT, Z-ST and FL A–C. GrouFailure loads after static loading are summarized in p = 0.714, nor a significant interaction between fatigue application and restoration material, F = 0.85, p = 0.469. However, the main effect of restoration material was significant, F = 19.20, p < 0.001. t-tests for the pairwise comparison between specimens with and without fatigue exposure for each material type confirmed this and showed no significance  = 0.14, ificance . The highest failure loads were observed for material FL both without prior fatigue application  and with prior fatigue application . Group RL showed the lowest failure loads of all materials tested, both without and with fatigue application. p < 0.05), except for the comparison of FL0 and Z-ST0 (p = 0.357) (p = 1.000) and between Z-HT and RL (p ≥ 0.197), regardless of fatigue application  . No difflication . p = 0.044) (p = 0.482). Materials Z-ST and RL showed a statistical difference in failure load with fatigue application (= 0.044)  but not All FDPs of Z-HT, Z-ST and FL1 revealed bulk fractures within the distal connector after single load to failure . FDPs ofZirconia ceramics and their different generations with increased translucency and beneficial esthetics, as well as the combination into strength-gradient multilayer pucks, are extensively investigated in current scientific research. In addition to zirconia, resin-matrix ceramics, such as PICN and CAD/CAM composites are widely analyzed due to their attractive application for chairside workflow and efficient machinability and processing . MoreoveThis laboratory study investigated the failure load and fatigue behavior of different designs of monolithic translucent zirconia, 3Y-TZP with a facial veneer and rapid-layer posterior FDPs bonded to one-piece zirconia implants. The tested null hypothesis was partially rejected as the type of restoration material affected the failure load. All tested materials survived dynamic loading up to 1.2 million cycles with simultaneous thermocycling, resulting in a 100% in vitro survival rate after chewing simulation. Measured failure loads (>2384 N) of all implant-supported FDPs surpassed normal physiological occlusal forces of 200–900 N in the posterior dentition, irrespective of fatigue application . AccordiTo the authors’ best knowledge, this is the first in vitro study that investigated different restorative concepts for FDPs on ceramic implants. Furthermore, no laboratory studies with monolithic all-ceramic FDPs on titanium implants are reported in the dental literature. Therefore, a comparison to other studies is difficult. Most in vitro studies investigated different posterior bi-layer 3Y-TZP FDPs on titanium implants ,43,44,45The highest failure loads both without and with fatigue application were measured for Group FL. Different from the monolithic groups Z-HT and Z-ST, group FL had a porcelain veneer over a 3Y-TZP framework. It is possible that t-m phase transformation due to moisture of the porcelain slurry may have led to compressive stresses, eventually increasing the load to failure . AnotherNo differences between failure loads of monolithic 3Y-TZP and 4Y-TZP FDPs could be detected. This is in line with a previous study in which no statistically differences could be found between failure loads of the same 3Y-TZP and 4Y-TZP material on SCs . TherefoThe lowest failure load values were measured for Group RL. As this might be expected with a PICN material as a tabletop on a 3Y-TZP framework, loads were nevertheless high. Most of the observed fractures would be repairable in a clinical scenario with either a new chairside fabricated PICN tabletop or a direct repair with a resin composite. The PICN material, with mechanical properties imitating a human tooth, such as a similar wear behavior and a dampening effect, might protect the implant from uncontrolled high occlusal forces and overloading . The resFailure and fractographic analysis of monolithic zirconia FDPs resulted in 100% bulk fractures with and without fatigue application. The fracture originated from the occlusal surface in the zone of the highest compressive forces down to the distal connector of the 3-unit FDP. As the contact area was on the pontic site and not along the axis of the implants, this unfavorable loading condition resulted in the gingival portion of the connector becoming the weakest link due to localized tensile stress with the highest probability of fracture and also an origin of a competing failure mode . For Group FL, almost all of the specimens showed the same fracture pattern with bulk fractures through the distal connector, except two specimens which showed bulk fractures through the pontic. Interestingly, the facial veneering withstood the high forces and did not fail before bulk fracture. However, Group RL showed a different failure pattern: 80% of the specimens showed chipping fractures of the PICN tabletops without detachment and involvement of the 3Y-TZP substructure. Only 20% of the RL specimens fractured through the distal connector as the other groups. According to the present findings all tested monolithic and bi-layer all-ceramic restorations can serve as 3-unit FDPs on one-piece ceramic implants, whereas facial veneered 3Y-TZP and monolithic translucent 4Y-TZP FDPs presented the highest load to failure values. A limitation of this study is that the obtained result of the implant-restoration complex might be only transferred to a certain limit to other available one-piece ceramic implants. Future studies should compare the proposed restorative rapid-layer concept with novel strength-gradient multilayer zirconia SCs and FDPs. Other materials, such as bioactive restoratives, should also be explored as potential prosthetic materials ,55. LikeThe following conclusions can be drawn based on the obtained results and within the limits of this laboratory study:The applied fatigue protocol had no effect on the failure load of the materials investigated.All tested prosthetic reconstructions showed higher failure loads (>2384 N) than normal physiological occlusal forces (200–900 N) in the posterior region and can be used clinically.Bi-Layer FL and monolithic Z-ST showed the highest resilience and might serve as reliable prosthetic reconstruction concepts for 3-unit FDPs on ceramic implants.The Rapid-Layer concept shifted failure modes from catastrophic bulk fractures to clinically repairable failures within the PICN veneer layer.The proposed rapid-layer design with a PICN “tabletop” might be an interesting restorative treatment concept for zirconia implants due to their tooth-like wear behavior and easy replaceability.Proper veneer design with a limited extension to the buccal/facial area resulted in superior failure load results.Future research should focus on novel strength-gradient multilayer zirconia ceramics for SCs and FDPs on one- and two-piece ceramic implants.


Highland barley (HB) is a nutritious crop with excellent health benefits, and shows promise as an economically important crop with diverse applications. Starch is the main component of HB and has great application potential owing to its unique structural and functional properties. This review details the latest status of research on the isolation, chemical composition, structure, properties, and applications of highland barley starch (HBS). Suggestions regarding how to better comprehend and utilize starches are proposed. The amylopectin content of HBS ranged from 74% to 78%, and can reach 100% in some varieties. Milling and air classification of barley, followed by wet extraction, can yield high-purity HBS. The surface of HBS granules is smooth, and most are oval and disc-shaped. Normal, waxy, and high-amylose HBS have an A-type crystalline. Due to its superb freeze-thaw stability, outstanding stability, and high solubility, HBS is widely used in the food and non-food industries. The digestibility of starch in different HB whole grain products varies widely. Therefore, the suitable HB variety can be selected to achieve the desired glycemic index. Further physicochemical modifications can be applied to expand the variability in starch structures and properties. The findings provide a thorough reference for future research on the utilization of HBS. Highland barley  is a variety of barley that belongs to the grass family and is an annual herb, which can be classified into two-, four-, and six-row HB based on the number of ridges, as well as white, black, and purple HB based on color. The characteristics of HB include a high degree of cold tolerance, a short growing period, wide adaptability, early maturity, and high yield, and it is suitable for cultivation in the cool climate of the plateau . In 2019In comparison with other cereal crops, HB had better nutritional value as it contains higher protein, vitamin, and fiber contents, especially β-glucan . β-glucaThe primary component of HB is starch, which accounts for 58.1–72.2% of the dry weight and generally comprises 74–78% amylopectin, and up to 100% in a few varieties . The strIn comparison to other common starches, HBS has not been analyzed systematically, which hinders the in-depth research of HB and the application of HBS. In this review, we aim to summarize the isolation methods, chemical composition, structure, properties, and applications of HBS. Additionally, future research suggestions are recommended.Physical techniques, enzymatic hydrolysis, and other procedures can be used to decompose the cell wall of HBS, which consists of cellulose, hemicellulose, and pectin. Following the release of the cellular contents, proteins can be separated using appropriate procedures to yield pure HBS. The current methods for extracting HBS are the dry, wet, and wet-dry combination approaches.Dry extraction is a useful process for enriching certain nutrients  in barley. Pearling ,24, rollWet extraction of HBS frequently requires the use of alkaline and enzymatic techniques. Soaking with lye can degrade or loosen the protein around the HBS, weaken the HBS-protein combination, and dissolve the protein to obtain high-purity starch. Yang et al.  extracteThe enzyme cellulase is commonly used for starch extraction, and it acts on the cell wall of HB to rupture and disintegrate it, enabling the complete release of cellular contents, which is beneficial for the separation of starch and protein . ExtractThe presence of β-glucan is a concern, as it absorbs a considerable quantity of water during washing and increases the slurry viscosity, making it difficult to separate in subsequent steps. Technology has been developed for separating high-purity starch from HB grains with different amylose contents. It is possible to separate starch and fiber fractions from whole barley flour in a semi-aqueous medium (50% ethanol) without affecting the viscosity of β-glucan. The majority of the starch isolates thus obtained had high purity large particles, with yields ranging from 22–39%. More significantly, the extraction efficiency of the β-glucan component was 77–90%, indicating that separation from the starch component during processing was efficient .Barley was subjected to milling and air classification operations to identify the graded fractions with high starch content based on particle size and composition, and the fractions were further separated based on the wet technique. The combination of wet and dry approaches can minimize water consumption and centrifugal load while increasing starch yield. During the milling and air classification procedures, most of the β-glucan is transferred to other components, which is advantageous for further separation and purification of starch.There are significant differences in the chemical composition of HBS . Amylose7 g/mol. The molecular weight of HBS was greater than 4 × 107 g/mol, and was mainly distributed as 4–6 × 107 g/mol, 6–8 × 107 g/mol, and >8 × 107 g/mol fractions, with distribution ratios of 32.26%, 38.41%, and 29.33%, respectively. Naguleswaran et al. . Ho. Ho55]. −1, which corresponds to the crystalline-amorphous lamellar structure of the starch granule, and the thickness of the lamellar structure is around 9.33 nm [Small-angle X-ray scattering (SAXS) is commonly used to characterize the lamellar structure of starch granules. The amylopectin side chains are arrayed in parallel to form a double-helical crystalline region with a high electron cloud density, whereas the branched regions of amylopectin form an amorphous region in the lamellar structure. Together, these two regions form a lamellar structure of 9–10 nm. The characteristic peaks from the SAXS analysis of starch are formed because of the differences in electron cloud density in different regions of the layered structure. HBS has a prominent scattering peak at ~q = 0.6 nm 9.33 nm ,60.The crystal orderliness of starch granules includes the long-range orderliness of the neatly packed double helix and the short-range orderliness of the double helix packed at short distances from each other. The short-range ordered structure, as part of the long-range ordered structure, is a prerequisite for the existence of long-range ordered structures. Short-range ordered structures are less susceptible to disruption than long-range ordered structures, and their presence does not always imply the formation of long-range ordered structures . The FouThe gelatinization properties of starch are essential indicators of its quality. The gelatinization temperature of starch can be determined by DSC , thermomThe swelling power of starch is expressed as its capability to absorb water during gelatinization, and the paste’s capability to hold water after centrifugation . The masRapid visco-analyzer (RVA), Brabender viscograph, and rheometer can be used to analyze the pasting characteristics. RVA is more commonly used technique due to the small sample size required and fast detection speed. Several raw materials used in food machining are fluids, and their rheological characteristics are modified by the temperature and concentration of the food material, which in turn affects processing properties, such as food output, mixing, and agitation. Cereal powders, such as starch, are frequently utilized as pastes, and the rheological properties of the paste can influence product qualities, including viscosity, hardness, and texture. Typical starch pastes exhibit non-Newtonian shear-thinning fluid characteristics . Zhu et During the process of freezing gelatinized or ungelatinized starch at low temperatures  and subsequently thawing it at normal or higher temperature  to melt the starch, the changing trend and degree of starch physicochemical properties and granular structure reflects the freeze-thaw stability of starch, which has a direct impact on the textural attributes of associated quick-frozen foods. The amylose content affects freeze-thaw stability, with high amylose content resulting in poor freeze-thaw stability. Studies have shown that after four freeze-thaw cycles, there was a 4% syneresis for zero amylose HBS, compared to 21% for CDC Candle HBS with 5% amylose . Zheng eThe degree of light transmittance after starch is completely formed into a paste is called the transparency of starch paste, which can be expressed as light transmittance. Greater light transmittance correlates with higher transparency. The transparency of starch paste is positively associated with the amylopectin content . The solThe viscoelasticity and gel strength affect the machining and molding of the gel, as well as its flavor and quality. The mechanical taste of starch gel changes the viscoelasticity, hardness, and roughness of food, which is different from the chemical taste caused by sugar, inorganic salts, acid, alkali, etc. Gel strength varies with temperature; however, this change is reversible. During storage, amylopectin recrystallization is the major cause of the increase in gel hardness . The preBased on the speed of starch digestion, starch is nutritionally classified as rapid digest starch (RDS), slow digest starch (SDS), and resistant starch (RS) . StudiesHBS contained RS, SDS, and RDS within ranges from 13.1–31.6, 19.8–25.7, and 14.4–19.6 g/100 g dry starch, respectively . Shen etHB contains an average of 5.25% β-glucan, which inhibits the functions of α-glucosidase, α-amylase, and invertase, thereby affecting the digestibility of HBS . With hiUsing appropriate chemical, physical, and enzymatic modification techniques, the particle, lamellar, crystalline, amorphous, and chain structures of starch can be regulated to modulate starch digestibility and confer diverse nutritional benefits. Currently, the United States, Japan, Australia, and other countries have developed commercial anti-digestive starch products, some of which have up to 90% anti-digestibility properties .Based on its unique structure, properties, and chemical composition, HBS is especially suited for use in food and non-food applications. The high water holding capacity and salt-resistant gelatinization stability of HBS make it useable in meat products and as a soup thickener. Shand  found thOur knowledge of the properties of HBS has progressed significantly. The main component of HB is starch, which accounts for 58.1–72.2% of its dry weight. The starch obtained by milling and air classification of barley, followed by wet separation, is of high purity. Several studies have shown significant differences in the chemical composition, particle and molecular structure, and physicochemical characteristics of HBS. In general, HBS granules have a smooth surface, and most are oval or disc-shaped. Normal, waxy, and high-amylose HBS have an A-type crystalline form with degrees of crystallinity ranging from 10.72–43.21%, 33.0–37.1%, and 29.1%, respectively. The pasting properties of distinct highland barley starches vary and are influenced by the purity of the starch, amylose/amylopectin ratio, amylose and amylopectin content, and granule size. HBS is widely used in food and other industries because of its outstanding freeze-thaw stability, high solubility, excellent emulsifying stability, and favorable stability. Non-starch components are key factors contributing to the low glycemic index of HB-based foods. The variability in starch structures and properties can be further enhanced by physical and chemical modifications. To maximize the use of HBS, it is necessary to consider the relationship between chemical composition, structure, properties, modifications and applications, not just from a one-sided perspective.


Beam hardening artefacts induced by highly-dense material  is a common quality issue in maxillofacial Cone Beam Computed Tomography (CBCT-) images. This experimental and analytical study investigated attenuation patterns of two typical dental implant materials: zirconia-ceramic and pure titanium. By application of different x-ray beam energies  energy-dependent attenuation of these materials is assessed and the resulting artefact induction in the resulting CBCT-images evaluated. A zirconia (Y-TZP-) implant ( As implants have become a common restoration in many countries, often they will also be depicted in CBCT-scans acquired from their carriers. The vast majority of dental implants is made of highly pure Titanium with a chemically very stable oxide-surface2. Their artefact influence in CBCT scans has been experimentally described5 and also analytically explained6. In recent years, dental implants made of zirconia are increasingly marketed and inserted7. These are made of crystalline zirconium dioxide generally stabilized with 3 mol% Yttrium (Y-TZP)8. The dimensions and designs are roughly the same as in titanium implants. The fundamental part of beam hardening artefacts occur due to the filtering effect of the highly-dense, i.e. severely attenuating implant body which changes the energy-spectrum of the x-ray beam. After penetration of such highly-dense (strongly attenuating) bodies the beam contains relatively more higher-energetic (shorter wavelength) x-rays than the spectrum emitted by the source. This process is termed beam hardening. Unfortunately, the reconstruction assumes identical energies in the spectra and this error propagates into the three-dimensional (3D) reconstruction process9. Briefly explained, the relatively too high energy arriving at the detector “behind” the attenuating object (here: the implant) is being backprojected into the reconstructed volume, resulting in dark (hypo-dense) lines in beam direction. Attenuation is mainly caused by two main mechanisms: the Compton scattering and the photoelectric effect10. While the former is rather stable over the energy spectrum typically applied for CBCT, the photoelectric effect is strongly energy-dependent10. Here, the energy of the beam is capable to remove an electron from the inner k-orbit thereby producing a ion with a positive charge. At energies just above the required energy to remove the electron from the k-orbit of the respective material there is an abrupt increase of the attenuation of the material. This so-called “k-edge” is material-specific and increasing with increasing atomic number. The k-edge attenuation energy of the two materials investigated here is 4966.4 eV for Titanium versus 17,997.6 eV for Zirconium as main compound of zirconia-ceramic. Empirical studies investigating artefacts in CBCT-images caused by zirconia have been published13. However, to better understand the background and to potentially minimize their effects, these artefacts should also be studied from an analytical perspective using physical data and knowledge on the 3D reconstruction process.Cone Beam Computed Tomography (CBCT) plays an important role in preoperative planning of dental implantsDespite the rather large body of literature existing for experimental data on titanium and zirconia-ceramic (Y-TZP) CBCT-artefacts, to the best of our knowledge no analytical data on the underlying attenuation differences and their effect on the artefacts for different beam energies have been published so far. This work intended to discuss the background and investigate the artefact burden arising from zirconia-(Y-TZP-) and titanium-implants when imaged in a CBCT-machine using different beam energies. For this purpose, from the projection radiographs used for CBCT-reconstruction, the actual attenuation was computed for both materials and four different beam energies. The results of the attenuation experiments subsequently were compared to the beam hardening artefacts measured in the reconstructed implant images.9. Titanium with an atomic number of 22 has been shown to considerably alter the x-ray spectrum for the typical energies (60–120 kVp) used in dentomaxillofacial CBCT-machines6. Zirconium as base-substance for zirconia-implants has an atomic number of 40, hence inevitably the beam-hardening effect will be more prominent when compared to titanium. Y-TZP-implants are composed of zirconium- and yttrium-oxide. Unfortunately no exact mass-attenuation coefficients are available from the National Institute of Standards and Technology Web site (physics.nist.gov). Hence, for an analytic evaluation on expectable artefacts caused by Y-TZP experimental attenuation measurements are required. We will derive the attenuation from zirconia and titanium from the 2D projection radiographs (PROJs)  the filtering object  significantly differs from the emitted beam in composition. Since it only contains the shorter (higher energetic) wavelengths, the beam is “hardened”. Unfortunately, the reconstruction process assumes equality between emitted and beam arriving at the detector. This discrepancy induces error in the reconstruction process visible as darker (combined with some brighter) lines and stripes that are always orientated in the direction of the x-ray-beam. The latter is the the opposite (inverse) direction of the reconstruction process which is based on (filtered) backprojectionrgies 60–20 kVp usI and x the distance the x-rays traverse the absorber. T, where r positioned at a focal-object-distance f (known from specifications of the CBCT-manufacturer) from the point-shaped x-ray source o that is at a distance h from the detector D  radiation, neglecting attenuation by air we assume that emitted radiation equals the incident radiation in areas where only air is depicted. The linear attenuation coefficient is obtained from:entclass1pt{minimaentclass1pt{minimaentclass1pt{minimax the beam passes through the implant  an Y-TZP circonia implant  of 4.1 mm diameter and a pure titanium rod  of diameter 4.0 mm. All exposure parameters  were extracted using ImageJ (https://imagej.nih.gov/ij/download.html) and saved in original 16 bit depth  for all four energies was relatively equal and only around 5% of x within the implant is very similar for all beam energies. Regarding attenuation, for the 4 mm thick material a Y-TZP attenuated 68% more than titanium at 60 kVp, versus 225% more at 90 kVp  contains a large amount of x-rays in this energy range between 4 keV and 17 keV. From Fig. 15. This work provides an analytical evaluation on the artefact-level caused by zirconia-implants (Y-TZP) versus pure titanium implants. Y-TZP composition as typical zirconium-oxide material with Yttrium-oxide additive was selected as representative for zirconia-implants. By using the 2D radiographic projection images acquired during a CBCT-scan as input for the assessment and the mathematical calculations, the attenuation and transmission caused by typical implant diameters  was assessed and related to kilovoltage and material. For the typical peak kilovoltages (60–90 kVp) employed in dental CBCT-machines, up to 225% more attenuation of the x-ray-beam were observed for Y-TZP compared to titanium for which mass attenuation coefficients can be found in the National Institute of Standards and Technology website (physics.nist.gov) database. Unfortunately there is no such information easily available for zirconia. Hence, for a pure mathematical approach physical data are lacking and have to be derived from appropriate experiments. The approach of this experimental study was to use a subsample of the several hundred PROJs acquired in each CBCT-scan as input for the assessments. The rational behind this was that the manufacturers use these radiographs for the 3D reconstruction of the CBCT-volume. Hence it seems safe to assume that not overly much processing of these input images  should be conducted. At least, it seems very likely that only linear operations are applied since the densities (representing energy incident on the detector) measured on the detector-pixels directly propagates into the inverse radon transform applied for 3D-reconstruction. Under this assumption the range between “no attenuation” as represented by only air images on the PROJs and the attenuation in the shadow of the implant was be defined as intensity range. Albeit physically not really correct, in the light of an unknown spectrum emitted by the source the simplified model used in this study allows for a reasonable modeling of the emitted intensity. Since the acquisition geometry of all PROJs was identical, the error induced by this simplification will be small. Plus, since the same error enters the equation . These findings are also supported by the literature. Vasconcelos et al. for 70 kVp, 80 kVp and 90 kVp found significantly pronounced artefacts for zirconia when compared to a titanium implant17. For identical energies, the same relationship was reported by Freitas and colleagues18. From the experimental data it can be concluded, that beam hardening for Y-TZP in comparison to Titanium cannot be markedly reduced by higher voltages up to 90 kVp. This finding is also in agreement with the evaluation of the reconstructed CBCT-data  along the projection (reconstruction) lines in the reconstructed volume. Clinically spoken, dark streaks occur along this direction which often compromise readability of the CBCT-images. The higher the atomic number, the more beam hardening will occurvoltages 0–90 kVp voltages 0–90 kVp From the experiments and the mathematical modeling of the attenuation process we observed that Y-TZP-zirconia implants in the typical energy ranges employed in dental and maxillofacial CBCTs (60–90 kVp) will attenuate up to 225% more beam-energy when compared to pure titanium implants. For the energy spectrum used in this study, beam hardening caused by titanium can be reduced using higher energies while this is not the case for zirconia-ceramic (Y-TZP).


Brassica rapa) yield and quality. To precisely characterize the Plasmodiophora brassicae infection of Chinese cabbage, we developed a dual fluorescent staining method for simultaneously examining the pathogen, cell structures, and starch grains. The number of starch (amylopectin) grains increased in B. rapa roots infected by P. brassicae, especially from 14 to 21 days after inoculation. Therefore, the expression levels of 38 core starch metabolism genes were investigated by quantitative real-time PCR. Most genes related to starch synthesis were up-regulated at 7 days after P. brassicae inoculation, whereas the expression levels of starch degradation-related genes were increased at 14 days after inoculation. Then, genes encoding the core enzymes involved in starch metabolism were investigated by assessing their chromosomal distributions, structures, duplication events, and synteny among Brassica species. Genome comparisons indicated that 38 non-redundant genes belonging to six core gene families related to starch metabolism are highly conserved among Arabidopsis thaliana, B. rapa, Brassica nigra, and Brassica oleracea. Previous genome sequencing projects have revealed that P. brassicae obtained host nutrients by manipulating plant metabolism. Starch may serve as a carbon source for P. brassicae colonization, as indicated by histological observations and transcriptomic analysis. Results of this study may elucidate the evolution and expression of core starch metabolism genes and provide researchers with novel insights into the pathogenesis of clubroot in B. rapa.Clubroot is one of the major diseases adversely affecting Chinese cabbage ( Plasmodiophora brassicae, is a serious global threat to field-grown Brassica species. After infection by P. brassicae, plant roots often form galls, whereas the aboveground parts turn yellow, which adversely affects the final yield. Clubroot is very difficult to prevent and control because P. brassicae resting spores remain viable in the soil for >17 years in the absence of host plants [Brassica plants. Clubroot management has been based on the following two main strategies: preventing the pathogen from entering pathogen-free fields, and developing resistant cultivars [P. brassicae. Although the life cycle of the pathogen has been reported [P. brassicae, including some cruciferous weeds (stinkweed and shepherd’s purse) [P. brassicae and what genomic commonalities need to be understood.Clubroot, caused by a soil-borne biotrophic pathogen, ultivars , 3. Therreported –6, how is purse) . There iP. brassicae life cycle because it can elucidate the changes in various substances that occur during the infection of plants by P. brassicae. A histochemical analysis of plant tissues can elucidate the precise life phases of P. brassicae. Many staining techniques have been developed to identify pathogens, host structures, and changes in host cells during P. brassicae infections, including those utilizing methylene blue, astra blue, safranin, azure II, toluidine blue, 4′,6-diamidino-2-phenylindole, basic fuchsin, and Nile red alone or combined with other stains [P. brassicae and host cells in the late infection stages. However, during the early infection stage, it is difficult to distinguish between plasmodia and starch grains in host cells [Cytological research is important for clarifying the r stains , 10. Mosst cells , 12. AccBotryosphaeria dothidea and Botrytis cinerea, respectively [Arabidopsis thaliana roots infected by P. brassicae, but there has been no further research on the effects of P. brassicae on host plant starch metabolism [Brassica rapa also indicated that infected cells and clubroot galls contain more starch grains than healthy cells and roots [A. thaliana [B. rapa infected by P. brassicae. The core genes involved in starch metabolism are crucial for P. brassicae’s potential for manipulating plant metabolism to take nutrients from the host [. Fortunately, the available genome information can be used to identify the starch metabolism genes in Brassica crops (http://brassicadb.cn/). The relevant genomic distribution, structure, and duplication of these core genes as well as the syntenic analysis of B. rapa (AA genome), Brassica nigra (BB genome), and Brassica oleracea (CC genome) are important to reveal the commonalities of Brassica crops susceptible to P. brassicae [Starch serves as a transient and long-term carbohydrate reserve in plants and other eukaryotic organisms . Starch ectively , 18. Pretabolism , 20. Prend roots , 22. Genthaliana , 24, potthaliana , 26, andthaliana , 28. Litthe host –31. Fortrassicae .P. brassicae in infected and uninfected Chinese cabbage roots. The amylopectin level increased in B. rapa roots infected by P. brassicae. The expression profiles of the core enzyme genes related to starch metabolism were analyzed to reveal the P. brassicae-induced starch metabolism regulatory network in B. rapa. We also identified core genes encoding the following enzymes participating in starch metabolism in A. thaliana, B. rapa, B. nigra, and B. oleracea: ADP-glucose pyrophosphorylase (AGPase), starch branching enzyme (SBE), starch synthase (SS), starch debranching enzyme (DBE), α-amylase (AMY), and β-amylase (BAM). The chromosomal distribution, structure, duplication, and synteny of these identified genes were investigated. The results of this study may provide researchers with a comprehensive atlas of the cytological characteristics and starch metabolism during P. brassicae infection of B. rapa. The presented data will be useful for future studies aimed at increasing our understanding of the regulation of starch metabolism and the evolutionary divergence of these starch metabolism genes related to P. brassicae infections of the Brassicaceae family.In this study, we developed a dual fluorescent staining method useful for investigating the cytological characteristics of host cells and P. brassicae, cell structures, and starch grains, we developed a dual fluorescent staining method using aniline blue and Nile red. The isolated P. brassicae resting spores were stained blue , whereas the plasmodia were undetectable in uninfected roots ; SS contained the glyco_transf_5 domain (Pfam: PF08323); the debranching enzyme (ISA) included the CBM_48 (Pfam: PF02922) and α-amylase (Pfam: PF00128) domains; and all SBEs had the α-amylase_C (Pfam: PF02806) and α-amylase (Pfam: PF00128) domains. Of the enzymes in the starch degradation pathway, AMY contained the α-amylase_C2 (Pfam: PF07821) and α-amylase (PF00128) domains, whereas β-amylase (BAM) contained the glyco_hydro_14 domain (Pfam: PF01373)  .B. rapa genome  assay. The differences in the gene expression profiles suggested that the gene families were differentially regulated during P. brassicae infection of Chinese cabbage. The expression levels of the majority of the genes related to the starch synthesis pathway were up-regulated at 7 DAI. Specifically, BrAGPS2 and BrISA2b expression levels were significantly up-regulated at 7 DAI and gradually decreased from 14 to 35 DAI , after which they gradually decreased , B. nigra (BB genome), and B. oleracea (CC genome) is important to reveal the commonalities of Brassica crops suitable for the invasion and reproduction of P. brassicae. We therefore used the same approach to analyze the starch metabolism genes in B. nigra and B. oleracea, which are genetically closely related to B. rapa. A total of 38 (B. nigra) and 43 (B. oleracea) candidate genes from six core gene families related to starch metabolism were identified. Genomic distribution analysis demonstrated that 30 genes (100%) in A. thaliana, 38 genes (100%) in B. rapa, 36 genes (94.74%) in B. nigra, and 42 genes (95.45%) in B. oleracea mapped randomly and unevenly on the chromosomes   . On the basis of the phylogenetic analysis, the BAM genes were divided into four groups  . Based oleracea) . Four ISotif 10) . Fourteer groups . AccordiB. rapa genome. More specifically, 18 of 38 starch metabolism genes (47.36%) were associated with segmental duplications , B. nigra (59 pairs), and B. oleracea (64 pairs)  . Of the  BrBAM10 . Hence, Ka/Ks ratio is important in estimating selection pressure on genes during plant evolution. The analysis of Ka/Ks ratios for orthologous genes among four species showed that the ratios of segmental duplications and all orthologous gene pairs were <1 , B. nigra (BB genome), and B. oleracea (CC genome), we found that almost all genes related to starch metabolism in B. rapa have an ortholog in the other three species; the exception was BrBAM10. Most of the starch metabolism genes were single-copy genes with syntenic relationships among the A. thaliana and the A, B, C genomes. The exceptions were AGPL1, AGPS1, and ISA2, which were duplicated and homologous in Brassica species, implying the duplication events for these genes occurred after the A. thaliana lineage separated from the Brassica ancestor (43.2 million years ago) [Brassica species than in A. thaliana. In B. oleracea, three genes , which are not homologous to A. thaliana genes, encode the glyco_hydro_14 domain and are considered to be members of the BAM gene family. This may be related to a base sequence change during evolution, which resulted in the neofunctionalization of genes. Finally, the Ka/Ks ratios were all <1, indicating that these starch metabolism-related genes were under negative selection pressure during B. rapa evolution.To further clarify the molecular basis of starch accumulation, members of six starch metabolism gene families were identified in Chinese cabbage and classified based on comparative genomics and phylogenetic relationships. By analyzing the homology among genes from ars ago)  and befoars ago) . AdditioB. rapa, B. nigra, and B. oleracea can be explained by the triangle of U [A. thaliana is a member of the family Brassicaceae and has a common ancestor with Brassica species. Therefore, in the current study, with reference to A. thaliana genes, we systematically analyzed the six core enzyme gene families related to starch metabolism in Brassica crops, with a specific focus on B. rapa. Comparative analysis of orthologous pairs of starch metabolism genes in the four species showed that 36 (97.30%) genes in B. rapa corresponded to genes in the genomes of A. thaliana, B. nigra, and B. oleracea. These results indicated starch metabolism genes were highly conserved in the four cruciferous species during evolution.The genetic relationships among gle of U . FurtherBrassica crops identified several differentially expressed genes associated with the starch metabolic pathway that may influence plant responses to P. brassicae [P. brassicae infection of A. thaliana, SS genes are more highly expressed at 10 days after inoculation than at 23 days after inoculation [BrAGPS2 and BrISA2b, were significantly up-regulated at 7 DAI with P. brassicae. The expression levels of starch degradation-related genes were significantly up-regulated at 14 DAI, including BrBAM4a, BrBAM4b, and BrBAM8. The expression of these starch metabolism genes was positively correlated with the starch content.Previous transcriptomics-based analyses of rassicae , 41–43. culation . In the P. brassicae with carbon and energy during the infection of B. rapa. After the host plant perceives the infection, specific genes related to starch synthesis are activated, leading to an increase in starch contents. When the infected plant detects the proliferation of P. brassicae, the genes related to starch degradation are activated to provide the pathogen with carbon and energy, which are used for proliferation. We observed that the expression patterns of two starch synthesis pathway genes (BrAGPS2 and BrISA2b) and three starch degradation pathway genes  were consistent with the starch contents. Therefore, we believe these five genes are the key genes mediating the response of B. rapa to P. brassicae. The findings of this research provide novel insights into the relationships among the core genes involved in starch metabolism in B. rapa infected by P. brassicae.Based on our results, we propose that the accumulated starch (amylopectin) provides P. brassicae, cell structures, and starch grains in infected Chinese cabbage. By combining the results of cytological and starch quantification analyses, we demonstrated that the starch content significantly increased from 14 to 21 days after the P. brassicae infection was initiated. Moreover, 38 genes encoding core enzymes contributing to starch metabolism were identified based on genome-wide analysis. The expression levels of genes participating in the starch synthesis pathway increased significantly from 7 to 14 days after plants were inoculated with P. brassicae. The expression of genes involved in starch degradation increased significantly after 14 days. Finally, according to syntenic analysis, the starch metabolism genes were revealed to be highly conserved in A. thaliana, B. rapa, B. nigra, and B. oleracea. The data presented herein may be useful for characterizing the evolution and regulatory effects of the core starch metabolism genes in Chinese cabbage. This study provides new insights into P. brassicae pathogenesis and interactions with Brassica plant hosts.In this study, we developed a dual fluorescent staining method applicable for distinguishing between A. thaliana ecotype Columbia (Col-0) were used in this study. ‘BJN3-1’ is highly susceptible to P. brassicae [The Chinese cabbage inbred line ‘BJN3-1’ and an rassicae .P. brassicae isolate ‘LNXM-8’, which was identified as pathotype Pb1 according to the Sinitic clubroot differential set, was used to inoculate ‘BJN3-1’ and Col-0 plants [et al. [0 plants . The pre [et al. . ‘BJN3-1P. brassicae or distilled water. Root samples of Col-0 were collected at 28 DAI with P. brassicae. Ten repeats of root cross-sections were prepared as previously described [The middle part of the main root of Chinese cabbage ‘BJN3-1’ and Col-0 plants was washed and placed in a 10% formalin solution used for preparing paraffin sections . Root saescribed . ParaffiP. brassicae or distilled water were collected at 3, 7, 14, 21, 28, and 35 DAI. Three replicates of 20 individual plants were used for quantifying starch using a commercial starch assay kit . For a qualitative analysis of starch, 10 repeat root cross sections prepared from samples collected at 21 DAI along with amylose and amylopectin from potato were stained with a fresh iodine solution (I2/KI) for 3 minutes and then washed with distilled water for 30 seconds.Chinese cabbage roots inoculated with A. thaliana genome were retrieved from the TAIR10 database (https://www.arabidopsis.org/). Additionally, B. rapa, B. nigra, and B. oleracea genome assembly and gene annotation data were downloaded from the BRAD database (http://brassicadb.cn/). We identified the starch metabolism genes according to sequence similarities and specific profile HMMs. To determine sequence similarities, a BLASTP analysis was used to identify the predicted starch metabolism genes, using A. thaliana genes as queries and the following criteria: e-value <1e−20, identity >70%, and match length > 70% (of the shorter protein sequence). Regarding the profile HMM approach, domains in the A. thaliana starch metabolism genes were detected according to the Pfam 33 database. We then used hmmsearch of the HMMER program (version 3.3) [B. rapa, B. nigra, and B. oleracea genomes. We confirmed the homologous relationships among these starch metabolism genes. Multiple gene sequences were aligned using CLUSTALW [Sequences of the annotated ion 3.3)  and the CLUSTALW  and phylCLUSTALW . FinallyP. brassicae or distilled water. Three replicates of 20 individual plants were used for the subsequent RNA isolation as well as qRT–PCR analysis. Firstly, total RNA was extracted from infected and mock control ‘BJN3-1’ root samples using TRIzol™ reagent . Subsequently, cDNA was synthesized from the purified total RNA (2 μg) using SuperScript™ III Reverse Transcriptase . The Primer 3.0 online program was used to design the primers for qRT–PCR; the primer information is listed in −∆∆Ct method [Entire roots were sampled at 3, 7, 14, 21, 28, and 35 DAI with t method , and theB. rapa chromosomes based on physical location in the reference genome and TBtools software [B. rapa were detected based on MCScanX [Ka/Ks ratios of the duplication events were calculated by KaKs_Calculator (version 2.0) [All identified starch metabolism genes were mapped to the software  was used MCScanX  with defion 2.0) .B. rapa and the other three species . The parameters were as follows: e = 1e−20, u = 1, and s = 5 [In this study, we use MCScanX to detected orthologous gene events between nd s = 5 . TBtoolsThis study received funds from the Liaoning Natural Science Foundation (2021-MS-229), Liaoning Revitalization Talents Program (XLYC1807242), the Key Research and Development Program of Hubei province, China (grant numbers 2020BBB083 and 2020BBA037), the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries through the Golden Seed Project, which is funded by the Ministry of Agriculture, Food and Rural Affairs (213006-05-5-SB110). These funding organizations had no role in the planning of this study, collection, analysis, interpretation of the data, and in the writing of the manuscript.Y.W., Y.B.M., and W.X.P. carried out the experiments and generated the data. Y.B.M., SRC, and WXP analyzed all data and wrote the original manuscript. X.Z., Y.J.W., X.Y.Z., M.Y.Z., D.L., Z.N.Z., D.Z., and X.Y.Y. participated in plant growth and dual fluorescent staining method development. C.X.G., S.S.C., and Y.L. participated in writing and modifying the manuscript. W.X.P., Y.P.L., and Y.L. conceived the study, participated in its coordination, and corrected and modified the manuscript. All authors have read and approved the final manuscript.Arabidopsis and Brassica genome databases used to generate data of this study are available in TAIR (https://www.arabidopsis.org/) and BRAD (http://brassicadb.cn), respectively.The data generated herein to support the results of this study are presented in the paper and the supplementary information files. The data involved in the phenotypic study are accessible from a corresponding author upon request. The The authors declare no competing interests.Horticulture Research online.Web_Material_uhab071Click here for additional data file.


Fermented wheat bran was produced by solid-state fermentation method, and the changes of nutritional indexes before and after solid-state fermentation were analyzed. Then, Arbor Acres (AA) broilers were used to study the effects of fermented wheat bran on growth performance, serum immune function, intestinal morphology and microflora of broilers under bacterial lipopolysaccharide stress. The present results indicated that the nutritional value of wheat bran was improved by solid-state fermentation, and wet FWB improved the immune profile of broiler chicken under stress.p < 0.05) average daily feed intake of days 21 to 42, and increased (p < 0.05) the villus height and villus height to crypt depth ratio of ileum on day 21, decreased (p = 0.101) the jejunum crypt depth and decreased (p < 0.05) the Lactobacillus and Bifidobacterium counts of the cecum digesta on day 42. Compared with NC, FWB-II and FWB-IV significantly increased (p < 0.05) the levels of serum total protein and globulin on day 21; compared with the basal diet groups, dry and wet FWB groups significantly increased (p < 0.05) glucose levels on day 21, and wet FWB significantly decreased (p < 0.05) alanine aminotransferase levels on day 42. Compared with PC and FWB-II, FWB-IV significantly increased (p < 0.05) the level of serum immunoglobulin G on day 21. Compared with PC and FWB-II, FWB-IV significantly decreased (p < 0.05) the levels of serum pro-inflammatory cytokines interleukin (IL)-6, IL-8, IL-1β and acute C reactive protein (CRP) on day 21; compared with FWB-III, FWB-IV significantly decreased (p < 0.05) the levels of IL-6, IL-8, CRP and tumor necrosis factor alpha on day 42, but the levels of IL-4 and IL-10 were significantly increased (p < 0.05) on days 21 and 42. These results indicated that supplementing 7% dry or wet FWB can improve growth performance and serum immune functions of broilers, which effectively alleviate the LPS-challenged damage, and wet FWB had a better effect than dry FWB.The study evaluated the effects of dry and wet solid-state fermented wheat bran (FWB) on growth performance, immune function, intestinal morphology and microflora in lipopolysaccharide (LPS)-challenged broiler chickens. The experiment was designed as a 2 × 3 factorial arrangement. A total of 252 one-day-old Arbor Acres male broiler chickens were randomly allocated to 1 of 6 treatments: basal diet + sterile saline , basal diet + LPS , 7% dry FWB + sterile saline (FWB-I), 7% dry FWB + LPS (FWB-II), 7% wet FWB + sterile saline (FWB-III) and 7% wet FWB + LPS (FWB-IV), with containing 6 replicate cages/treatment and 7 broiler chickens/cage, and the experimental period lasted for 42 days. Broilers were intraperitoneally injected with either 0.5 mg LPS or sterile saline solution per kg body weight at 16, 18 and 20 d of age. Growth performance, serum immunological parameters and indicators related to intestinal health were analyzed on days 21 and 42. Compared with NC, dry and wet FWB significantly increased ( Salmonella colonization and reduced heterophil to lymphocyte ratio; the functional compounds in fermented soybean meal, such as Lactobacillus plantarum, organic acid, short chain fatty acid and digestive enzymes, improved the intestinal morphology characteristics and immune response of young broiler chickens infected by Salmonella typhimurium. Similarly, Ashayerizadeh et al. [Salmonella typhimurium in the gut, decreased the heterophil to lymphocyte ratio and improved the growth performance of broiler chickens challenged by Salmonella typhimurium. Thus, fermented feed can be used as an alternative to antibiotics in animal production.In recent years, the immune stress caused by external pathogens seriously inhibit the growth potential of broiler chickens ,2. The eh et al.  reportedSaccharomyces and Lactobacillus increased the content of water extractable arabinoxylan by 3–4 times, increased the content and bioavailability of soluble dietary fiber and total free phenolic, and degraded over 20% of phytic acid. Additionally, adding 5% dry fermented wheat bran (FWB) to broilers (Ross 308) diets improved the growth performance, increased the Lactobacillus counts in ileum, and maintained the beneficial intestine environment [Wheat bran (WB) as the main byproduct of wheat starch processing, has rich nutrients and bioactive compounds ,12, suchironment . Zhang eironment , Lee et ironment  found thLactobacillus plantarum (LP17-1) and Bacillus subtilis (BS17-1) were obtained from Academy of National Food and Strategic Reserves Administration. Aspergillus fragrant (AF) was purchased from Wuhan Jiacheng Biotechnology Products Co., Ltd. . Saccharomyces cerevisiae (SC) was purchased from the Angel yeast Co. . LPS from Escherichia coli (E. coli) serotype O55:B5 (L2880) was purchased from Sigma-Aldrich Chemical Co. . The strains were prepared by shaking flask cultured with MRS broth, lysogeny broth and yeast peptone dextrose broth, respectively. Briefly, a 250 mL Erlenmeyer flask filled with 100 mL of different inoculum was covered with tin foil and autoclaved at 121 ± 1 °C for 15 min before use. Different microorganism was transferred to the related agar plate and cultured at 37 °C for 24 h. The concentration of viable bacteria in suspension was detected by the plate count method [O55:B5 L280 was puSolid-state fermentation of wheat bran was performed in a facultative anaerobic solid-state fermentation equipment to produce FWB. Wheat bran was mixed with 80% sterile water, and added to a 10% mixture inoculum , 2% glucose, then fermented at 27–33 °C for 3 days and stirred regularly per day. FWB was dried by boiling fluidized bed at 55 °C to collect dry FWB. Both dry and wet FWB samples were sealed and stored at 4 °C to determine the physical and chemical parameters.2CO3 solution and vortexed to mix thoroughly. After reaction for 60 min, the absorbance was measured by UV-spectrophotometer at 765 nm, and an equation was obtained from the standard gallic acid (GA), standard graph was used to determine the phenolic compounds of each sample . The crude polysaccharide content was quantified by a modified phenol-sulfuric acid method [Wheat bran and wet FWB samples were freeze-dried and crushed through 40-mesh sieve. The total phenolic content was determined using the Folin–Ciocalteu reagent described by Wang et al. . Brieflyd method . Brieflyd method . The mand method . BrieflyThe animal experiment was approved by the Animal Care and Use Committee of Academy of National Food and Strategic Reserves Administration (registration number: 2018L02). The animal experiment was performed at the Wuqing base of Academy of National Food and Strategic Reserves Administration .A total of 252 healthy one-day-old Arbor Acres male broilers (average body weight 43.8 ± 0.15 g) were purchased from a commercial hatchery , and randomly allocated to 1 of 6 treatments: basal diet + sterile saline , basal diet + LPS , 7% dry FWB + sterile saline (FWB-I), 7% dry FWB + LPS (FWB-II), 7% wet FWB + sterile saline (FWB-III) and 7% wet FWB + LPS (FWB-IV), with containing 6 replicate cages/treatment and 7 broiler chickens/cage. The experiment was designed as a 2 × 3 factorial arrangement with LPS treatments  and diet treatments . LPS was dissolved in sterile saline to prepare a concentration of 0.5 mg/mL. At 16, 18 and 20 d of age, birds were received an intraperitoneal administration of LPS or an equal amount of saline (0.5 mL).2). Birds were offered feed and water ad libitum. The temperature was maintained at 32 to 35 °C during the first week, which was progressively reduced by 2 to 3 °C a week until it reached 25 °C. The relative humidity was set at 50% throughout the trial. All chickens were vaccinated with an inactivated Newcastle disease-infectious bronchitis vaccine on day 7 and with infectious bursal disease vaccine on day 14. A starter diet (day 1 to 21) and grower diet (day 22 to 42) were designed to meet the nutrient requirements of China feeding standard of chicken (NY/T 33-2004) , LPS-challenged period (days 17 to 21) and later period (days 22 to 42). All chickens were housed in a temperature-controlled room with continuous light and breeding in (0.8 × 0.7 m33-2004) . All feeOn days 16, 21 and 42 of the experiment, one bird per replicate cage from each treatment group  was randomly selected and the body weight of all birds were recorded after an 8 h feed withdrawal. The growth performance was evaluated by final body weight (FBW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio , which was recorded on a cage basis during different periods.g for 15 min. Sera samples were separated and stored at −20 °C until biochemical analysis. Then the birds were euthanized by exsanguination and abdominal cavities were opened. The cecum digesta were collected in sterile EP (Eppendorf) tubes, then dipped immediately in liquid nitrogen and kept at −80 °C. Furthermore, 1 cm of the middle segments of jejunum and ileum were collected and fixed in 4% paraformaldehyde solution for later morphometrical assays.On days 21 and 42, blood samples were obtained in 5 mL vacutainer tubes from the brachial vein of birds, then centrifuged at 3000× The serum biochemical parameters, including glucose , triglyceride , cholesterol , alanine aminotransferase , aspartate aminotransferase , total protein , albumin  and globulin (GLB), which were determined by commercially available assay kits  and performed using an automatic biochemical analyzer . All procedures were performed according to the manufacturer’s instructions.β), tumor necrosis factor alpha (TNF-α) and acute C reactive protein (CRP) were determined by an ELISA method. ELISA kits were purchased from Beijing SINO-UK Institute of Biological Technology . All procedures were performed according to the manufacturer’s instructions.Serum immunoglobulin G (IgG), immunoglobulin A (IgA) and immunoglobulin M (IgM) were determined by immunoturbidimetry; interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-1 beta , cleared in xylene and embedded in paraffin wax. Sections of 4–5 μm thickness were stained with hematoxylin-eosin for general morphometry, according to Fischer et al. . The samE.coli was cultured with Aobox EMB agar under aerobic conditions at 37 °C for 48 h. Strains of Lactobacillus and Bifidobacterium were cultured with Aobox MRS agar and BS agar, respectively. The MRS agar plates and BS agar plates were incubated under anaerobic conditions at 37 °C for 48 h. The microflora numbers were calculated by plate count method, and expressed as log10 CFU per gram of cecum digesta [One gram of digesta sample was collected from the cecum. Samples were serially diluted with 9 mL phosphate-buffered saline solution and mixed thoroughly for enumeration of microbial populations.  digesta .p < 0.05 were considered statistically remarkable.The data was analyzed using SAS software . A student T-test was used to assess the effect of the fermentation process on the bioactive compounds and chemical composition of wheat bran. Data related to feeding period of broiler chickens were analyzed in a 2 × 3 factorial arrangement design using the GLM procedures of SAS software. The differences among treatments were evaluated by Duncan’s multiple range tests, and values of p < 0.05), 248.29% (p < 0.05) and 8.32% (p < 0.05), respectively. The contents of crude fat and crude fiber were reduced by 39.41% (p < 0.05) and 7.73% (p < 0.05), respectively.The nutrient contents of WB and FWB are presented in p < 0.001), 121.23% (p < 0.05), 58.77% (p < 0.001) and 95.65% (p < 0.05), respectively. The content of crude polysaccharide was increased by 78.88%.The major functional bioactive compounds are presented in p < 0.05) ADG and ADFI of broilers, while FCR was not significantly affected (p > 0.05) on day 16. During the LPS-challenged period (days 17 to 21), compared with NC, FWB-III significantly increased (p < 0.05) ADFI of broilers, all other parameters were statistically similar for all other treatments. During the later period (days 22 to 42), compared with NC, FWB-I and FWB-III significantly increased (p < 0.05) ADFI and FCR.The results of the growth performance of broiler chickens are presented in p > 0.05) in the LPS-challenged groups. Compared with PC, FWB-II and FWB-IV significantly increased (p < 0.05) the Vh and V/C in ileum. There was no significant difference (p > 0.05) between the dry and wet FWB groups. During the later period (days 22 to 42), the dry and wet FWB had a tendency to reduce (p = 0.101) the Cd of the jejunum.The results of the intestinal morphology of broiler chickens are presented in E. coli counts were significantly increased (p < 0.05) in LPS-challenged groups, but had no effect (p > 0.05) on the Bifidobacterium counts. Compared with the NC, FWB-III significantly reduced (p < 0.05) the Lactobacillus and Bifidobacterium counts, while E. coli counts were not significantly affected (p > 0.05) on days 42. During the later period (days 22 to 42), the E. coli counts of the cecum digesta were not significantly affected (p > 0.05) among treatments. Compared with the PC, FWB-II and FWB-III significantly decreased (p < 0.05) the Bifidobacterium and Lactobacillus counts of the cecum digesta; while FWB-IV significantly increased (p < 0.05) the Lactobacillus counts and decreased (p < 0.05) the Bifidobacterium counts, while E. coli counts were not significantly affected (p > 0.05). There was no significant difference (p > 0.05) between dry and wet FWB.The results of the cecum microflora numbers of broiler chickens are presented in p > 0.05) in serum biochemical parameters between the LPS groups and the saline groups. Compared with the NC, FWB-II and FWB-IV significantly increased (p < 0.05) TP and GLB levels. Compared with the basal diet groups, dry and wet FWB groups significantly increased (p < 0.05) GLU level. There was no significant difference (p > 0.05) between dry and wet FWB groups. During the later period (days 22 to 42), compared with the NC, FWB-III and FWB-IV significantly decreased (p < 0.05) the ALT level, while other serum biochemical parameters were not significantly affected (p > 0.05). Compared with the basal diet groups, the dry FWB groups had no significant difference (p > 0.05) in serum biochemical parameters, but the wet FWB groups significantly decreased (p < 0.05) ALT level.The results of the serum biochemical of broiler chickens are presented in p < 0.05), but immunoglobulin level was not significantly affected (p > 0.05) in LPS-challenged broilers. Compared with the PC and FWB-II, the FWB-IV significantly increased (p < 0.05) the levels of IgG on day 21. Compared with PC, FWB-IV significantly decreased (p < 0.05) the levels of pro-inflammatory cytokines IL-6, IL-8 and CRP, and the levels of anti-inflammatory cytokines IL-4 and L-10 were significantly increased (p < 0.05). During the later period (days 22 to 42), there was no significant difference (p > 0.05) in serum immunoglobulins among the diet treatments. Compared with FWB-III, FWB-IV significantly decreased (p < 0.05) the levels of pro-inflammatory cytokines TNF-α, IL-6, IL-8 and CRP, but IL-4 and IL-10 were significantly increased (p < 0.05).The results of serum immunoglobulins and cytokines of broiler chickens are shown in It was reported that solid-state fermentation (SSF) is an effective approach for improving the nutritive value of grain and oil processing byproducts by reducing the cellulose content and increasing a variety of biological active substances ,19. The Phytogenic-derived phenolic compound and polysaccharide can exert growth-promoting effect by its antioxidant function . He et aLPS is widely used to induce immune and oxidative stress models, which are characterized by anorexia, fever, impaired growth performance, and increased proinflammatory cytokine production for animals . In the These results suggested that the nutritional properties, bioactive compounds and flavor properties of wheat bran were improved by solid-state fermentation, which may promote the growth performance and immune function of LPS-challenged broilers. Moreover, direct application of wet FWB can not only reduce drying cost and bioactive compounds, but also be more conducive to growth-promoting effect ,8. ThereBased on our result, we can conclude that the basal diet supplemented with 7% dry or wet FWB significantly improved the growth performance and strengthened serum immune performance of broilers, which effectively alleviated LPS-challenged damage. Moreover, the potential of wet FWB was better in broilers immunomodulation.


Microcystis harmful algal bloom (HAB) using 3rd generation sequencing with the Oxford Nanopore MinION device. We classified the eDNA reads taxonomically, and estimated the abundances of all taxa in each sample. While the taxonomic data showed evidence of significant human and E. coli contamination, we found abundant Mycrocystis, especially in the samples drawn from bloom environments. The raw sequence data are available in the Sequence Read Archive (SRA) under accession number PRJNA812770. HABs pose a significant and increasing risk, both to human health and to the Blue Economy, and genomic approaches to early detection promise to help mitigate these risks. As such, this dataset could be of interest to freshwater ecology research teams, or any stakeholders interested in the detection and mitigation of HABs.Here we describe a publicly available environmental DNA (eDNA) sequence dataset, consisting of samples collected from a National Oceanic and Atmospheric Administration (NOAA) Great Lakes Environmental Research Laboratory (GLERL) on Lake Erie. We sequenced samples drawn from before, during, and after a 2019  Specifications Table•Microcystis bloom.The data provide a snapshot of the microbiota present before and during a •Microcystis cause significant harm to human health and local economies.Harmful algal blooms, including •Understanding the taxonomic makeup of the freshwater microbiome can provide insights into the dynamics of bloom formation.•Microcystis blooms specifically may find these data to be of value.Any stakeholders interested in the dynamics of HABs in general, and 1Microcystis bloom. We sequenced the DNA using the ONT MinION, and classified the reads taxonomically using the ONT What's In My Pot (WIMP) pipeline This dataset consists of raw environmental DNA (eDNA) reads from before and during a E. coli contamination present in several samples that was not revealed until taxonomic classification. While we took steps to track down and eliminate the contamination for later samples, we were unable to conclusively discover the source. Based on the quantities observed in the negative controls and reagent blanks, these included at minimum, 1,400 to several thousand reads classifying as E. coli, and from 50 to several hundred reads classifying as H. sapiens, and hundreds more classifying to other taxa, including Shigella, Acinetobacter, and Microcystis (though never more than 10 reads in any of the blanks or negative control). In some of the bloom-drawn samples (those from GLERL WE13) the abundance of E. coli was greater than that of Microcystis despite the fact that these were Microcystis blooms. Note that the abundance of Microcystis in the WE13 samples appears lower than in WE02, though still noticeably higher than in any of the pre-bloom or control samples. This could be in part due to lower bloom severity at the WE13 bloom during that sampling year. Additionally, the putative contaminants showed higher relative abundance in the WE13 samples and could have had a skewing effect on the abundance results.The results of the taxonomic classification are presented in www.glerl.noaa.gov). Details on the protocols for the collections at these sites are provided in the methods section.Collection sites are show in the map in Fig. 2 . SigSci is a subawardee recipient of funding from the NOAA SBIR grant noted above, which supported a proof of concept study to establish the technical merit, feasibility, and commercial potential of a technology.


The current review investigates outcomes and failure rates associated with arthroscopic circumferential allograft labral reconstruction of the hip, both as a revision and primary procedure in treating femoroacetabular hip impingement and labral-related pathology.Numerous studies within the last decade have demonstrated excellent patient-reported outcomes, high rates of return-to-play in athletes, and low failure rates in patients having undergone arthroscopic circumferential allograft labral reconstruction of the hip. Removal of chronically diseased and injured labral tissue can eliminate a significant pain-generator from the hip joint. Additionally, circumferential reconstruction of the labrum restores the hoop fiber strength and fluid seal akin to what would be seen with native, healthy labral tissue. Recent research has shown that arthroscopic circumferential allograft labral reconstruction may be used not only in the revision setting, but as a primary procedure. Circumferential labral reconstruction should be considered when a surgeon feels that the labrum is irreparable or has failed previous repair.Arthroscopic circumferential allograft labral reconstruction of the hip can be utilized as treatment option not only in revision settings, but also in primary treatment for femoroacetabular impingement and labral pathology This article will provide a brief overview of our progress as a specialty, evidence to support current practices as they pertain to arthroscopic circumferential allograft labral reconstruction of the hip, and implications for future directions in hip preservation surgery.While initially designed to resect damaged soft tissue, the goal of modern hip arthroscopy has shifted focus to preserving and restoring function within the hip joint and improving its biomechanics. Labral repair, which has been considered the standard treatment for hip impingement and labral tears, is becoming much more commonplace. However, considering a growing body of evidence demonstrating positive outcomes and low revision rates, we contend that labral reconstruction can be considered a primary treatment for femoroacetabular hip impingement (FAI) and labral tears, especially in such circumstances in which the labral tissue is deemed irreparable. A number of factors can contribute to poor healing with labral repair, thus, decreasing the labrum’s ability to provide a functional fluid seal around the joint, and increasing the likelihood of continued pain, intra-articular injury, and joint dysfunction . As suchDue to an exponential surge in demand, the last two decades have seen unprecedented growth within the specialty of arthroscopic hip surgery. Used as a technique to treat intra-articular pathology, including FAI and labral tears, hip arthroscopy has become increasingly common. Data from 2006 to 2010 demonstrated a 600% increase in the number of arthroscopic hip surgeries being performed annually in the USA , with a 40. Furthermore, the labrum functions to preserve intra-articular fluid pressure within the femoroacetabular joint [Within the last decade, a number of cadaveric studies have demonstrated the importance of the acetabular labrum in preserving normal hip function –41. Compar joint –40. Labrar joint –39. Biomar joint –39.While initially seen as a salvage procedure, our recommendation for primary labral reconstruction reflects the culmination of growing evidence supporting its benefit in revision settings and as a primary procedure to restore and preserve function when the labrum is deemed irreparable. This may include circumstances when labral tissue is compromised or otherwise inadequate or inappropriate for a repair, such as would be seen with congenitally or acquired labral deficiency, hypertrophic labra, ossified labra, or labral tissue that has been damaged, scarred, or compromised from previous surgery. In a 2020 cadaveric study, for example, Storaci and colleagues  found thIn a 2019 systematic review , Al ManaThe technique for labral reconstruction which we described in 2016 [First and foremost, the fundamental flaw with segmental labral reconstruction is that a shorter, roughly 4-cm graft is often placed in the anterosuperior quadrant which is the highest stress zone of the acetabulum. In this location and because the graft is short, it does not have surrounding attachments posteroinferiorly and anteroinferiorly. As it lacks surrounding support, this location of the segmental reconstruction can make healing and incorporation of the graft more challenging. This disruption of the circular structure of the labrum results in decreased hoop fiber strength of the remaining labral remnant. Unfortunately, there is no way to affix the segmental graft to the native labrum as the fibers run in parallel directions and suturing between the two structures often does not hold. Longer, circumferential grafts negate these issues as they span the distance from the origin of the anterior transverse acetabular ligament to the postero-interior acetabulum and cover all four quadrants of the acetabulum. Much like a suspension bridge, circumferential grafts are stronger. They provide rigid fixation antero-inferiorly and posteroinferiorly to give greater support to the critical, high-stress antero-superior acetabulum. In contrast, segmental grafts are placed in this zone of the acetabulum without surrounding support. The circumferential graft can reproduce the native fluid seal around the femoral head and thereby can more evenly distribute the forces associated with weight bearing and range of motion. Second, by removing only a segmental section of the labrum, the remaining, unhealthy, and highly innervated native posteroinferior and anteroinferior labral remnants remain in the joint and are vulnerable to further tearing and pain generation.23. Likewise, in reviewing literature on labral reconstruction over the past decade, we found that the majority of study protocols, especially within the last few years, utilized allograft tissue  . Our prePerhaps the most critical aspect of the procedure involves the meticulous treatment of the boney morphology causing the femoroacetabular impingement (FAI). During this stage of the operation, the femoral head neck junction (cam morphology) is reshaped to an anatomic, natural shape that will fit properly into the acetabulum. This eliminates impingement of the femoral head neck junction against the labral graft and improves both flexion and rotation of the hip. The acetabulum is circumferentially excoriated to prepare for biologic incorporation of the labral graft and pincer morphology, or over coverage, is removed when appropriate Figures  and 2. IWith both sides of the joint reshaped and prepared, suture anchors are placed measuring 10 to 12 mm apart around the entire acetabular rim . Anchors are placed from one of two direct anterolateral (DALA) portals, which allow access to both the anterior and posterior acetabulum. To avoid eversion of the labral graft, suture anchors should be placed as close to the cartilage border as possible. It is helpful to place all suture anchors before the acetabular graft is introduced into the joint to allow for optimal visualization and anchor positioning. After all suture anchors have been placed, and before the graft is introduced into the joint, two small drill holes measuring approximately 0.6 mm in diameter are placed between each anchor site. The drill holes create vascular channels which foster osseous integration of the graft into the acetabular rim and are particularly helpful in areas where only the acetabular edge could be excoriated to avoid loss of cup volume Figure .Fig. 3VGraft length is determined by measuring the length of the labral defect from front-to-back with a 4-mm-wide elevator. Approximately 3 to 4 cm are then added to the measurement to account for the nonlinear contour of the acetabular rim. After graft length has been calculated, the graft is meticulously prepared by tubularizing the fascial tissue. This is accomplished by folding the fascia into thirds or quarters and then passing a 2-0 Vicryl suture through several small bites in an accordion-like manner at the end of the graft. These are tied, and the suture tails are then attached to a graft master to maintain adequate tension on the graft. Using a 2-0 Vicryl suture, a baseball stitch is run from front to back through the graft, using intermittent circumferential wraps, to compress the tissue.The preferred diameter of the final graft varies among surgeons. A larger diameter graft is easier to achieve a seal with the femoral head but takes up space during the operation and is harder to compress and achieve incorporation. In contrast, a smaller diameter graft is easier to compress and incorporate, but it is harder to obtain a final seal between the graft and the femoral head. Over the years, we have found that a final graft measuring 5 to 5.5 mm in diameter provides adequate material to establish a seal with the femoral head and can be rigidly fixed and compressed. This size also appears aesthetically proportionally appropriate.Using a cannula, the graft is brought into the joint via the anterior DALA portal. A suture limb from the most antero-inferior anchor is tied directly to the graft in figure of eight fashion. With the graft fixed to the suture, the anchor is used as a pulley to advance the graft into the joint. Once in the joint, the graft is placed in provisional position around the acetabular rim with a probe. Once positioned, a probe through the anteromedial (AM) portal allows the surgeon to maintain tension on the graft. Sutures are secured for the first two to three anchors. The most difficult area to create a seal between the graft and the femoral head is the antero-superior zone as it represents a challenging transition for the rigid graft from the vertical anterior wall of the acetabulum to the horizontal/lateral zone of the acetabulum. To offset the potential of not having a seal between the graft and the femoral head in this section, the suture anchors are passed, but not tied, at the anterior-superior, lateral, and posterior positions around the acetabulum. The graft is then fixed posteroinferiorly to tension the graft so that it can follow the curvature of the antero-superior acetabulum.After all sutures have been passed, the graft is tensioned and cut posteroinferiorly. It is important to note that the graft is cut in the joint to ensure that its length is appropriate. This is the advantage of the Front-to-Back technique as graft length is tailored in situ to avoid a mismatch between the graft length and the length and contour of the acetabulum . For lonAt the most posteroinferior aspect of the acetabulum, two anchors are placed only a few millimeters apart. Using an Elite Pass (Smith & Nephew), the sutures from the distal anchor are passed through the graft and are then tied, while the sutures from the adjacent anchor are tied circumferentially around the graft. With the graft now fixed antero-inferiorly and posteroinferiorly, the sutures in between can be secured. This is done in the peripheral compartment with the hip joint reduced or off traction. The camera is then moved to the AL portal and a cannula is positioned in the posterior DALA portal. The hip is then taken off traction which reduces the graft to the rim of the acetabulum and relieves tension on the anchors. Remaining sutures are tied in the peripheral compartment to ensure rigid fixation of the graft. It is absolutely critical that the graft form a perfect seal around the femoral head Figure . Once thPublications reporting on the outcomes of arthroscopic allograft labral reconstruction have continued to increase, with eight new studies having been published in 2020–2021, alone, on the topic ,16,19,46n = 15 re-repair, n = 98 reconstruction) which had previously undergone previous labral repair or debridement5 for an average of 2.4 (reconstruction) to 4.7 (revision repair) years. Hips that underwent revision arthroscopy with labral revision-repair were 4.1 times  more likely to fail treatment when compared to patients who underwent revision arthroscopy with labral reconstruction  [Those who have failed a previous hip arthroscopy represent a challenging patient population and there has not been an evidence-based consensus on the best approach for them. As such, in 2016 we compared outcomes between patients who underwent revision hip arthroscopy with iliotibial band allograft labral reconstruction versus labral re-repair . In our In regard to labral reconstruction and evaluation of clinical outcomes, it is of salient importance to mention that the lead author performed over 3,000 arthroscopic allograft labral reconstructions between July 2009 and February 2020 — both as primary and revision procedures . In 2012p<.01) [In 2018, we published a self-controlled cohort, or case-crossover, study investigating the differences in outcomes between primary labral repair and primary labral reconstruction in the same patient, performed on contralateral hips. We believe this study has provided some of the strongest evidence supporting primary arthroscopic labral reconstruction of the hip . In our p<.01) . The patIn addition to differences in treatment failure rates, we found that patients who had undergone labral reconstruction noted superior outcomes compared to repair on a number of patient-reported outcomes, such as the mHHS, LEFS, VAS, and with average pain with activities of daily living (ADLs) . Additiop < .0001) and an average 27-point increase in postoperative LEFS (p < .0001).In a 2016 study [49,50. Likewise, another recent study found that hips with a Tönnis grade of more than 1 had as much as a 133% increased risk of converting to THA [In the area of hip arthroscopy, literature as it pertains to outcomes and age has focused on labral repair, not reconstruction. Two recent systematic reviews focused on hip arthroscopy outcomes in patients ages 40 years and older found that while there was an improvement in PROs the conversion to THA was as high as 30%g to THA . The rolg to THA . While wn = 158), labral repair in patients 40 years and older (n = 93), and a control group of labral reconstruction in patients ages 30 to 39 years (n=112) [p = .02), and that there was no difference in failure rates between the 30 to 39 reconstruction group when compared to the over-40 reconstruction group  [p < .01). For example, patients in the over-40 reconstruction group reported a 37-point improvement on the mHHS, whereas the over-40 repair group reported a 28-point improvement.As such, in 2020 we published a study comparing outcomes of patients over the age of 40 who underwent primary labral reconstruction or primary labral repair. Our hypothesis contended that aged and chronically diseased labral tissue is compromised and does not heal well with labral repair. Improved results have been demonstrated when this labral tissue is removed, and the patient undergoes a circumferential labral reconstruction. In our “over-40” study, we followed 312 hips for approximately 4 years [Considering the positive evidence supporting labral reconstruction, the lead author performs only this procedure and is an advocate for labral reconstruction as both a primary and revision procedure as a more complete solution in the treatment of labral tears and FAI in the presence of irreparable labral tissue. It should also be regarded as the standard for revision hip arthroscopy. However, this stance is considered by some to be controversial as they still relegate labral reconstruction to a salvage operation. The decision to reconstruct the labrum should be made based out of respect for the operation and surgeon experience. When performed poorly, labral reconstruction has the potential to be catastrophic, especially in instances where the acetabular rim is over-resected, thereby resulting in iatrogenic dysplasia. Conversely, when performed well, current evidence has demonstrated a high likelihood of success with labral reconstruction as a primary operation. In direct comparison studies to labral repair, the outcomes with labral reconstruction are similar or better depending on the institution.The field of hip arthroscopy has grown exponentially in the last decade, and as a specialty, we have seen substantial advancement in the technical abilities of surgeons, and the evolution of the operation itself. Labral reconstruction plays a vital role in hip preservation by re-establishing normal anatomy and function especially in situations where the native labrum cannot be preserved. When compared to labral repair, research has shown that labral reconstruction has the potential to provide more significant improvements in pain and restoration of function. Labral reconstruction should have a role in the practice of every high-volume hip arthroscopist. Current evidence supports our recommendation that arthroscopic circumferential allograft labral reconstruction should be performed in all revision settings and in any instance when surgeons believe the labrum is irreparable. As evidenced by several recently published studies, patients across a broad spectrum of indications who undergo labral reconstruction, whether as a primary procedure or in the revision setting, demonstrate improved outcomes and low rates of failure. While a number of surgical techniques, including a variety of allograft options, have been described in the literature, our research and review of literature supports our recommendation for a circumferential, front-to-back fixation using an iliotibial band allograft, where the graft is measured and cut inside the joint , 15. The