H-pyrazole (EH7) showed the highest potency against MAO-B with an IC50 value of 0.063 µM. The potencies against MAO-B were increased in the order of –F (in EH7) > –Cl (EH6) > –Br (EH8) > –H (EH1). The residual activities of most compounds for MAO-A were > 50% at 10 µM, except for EH7 and EH8 . EH7 showed the highest selectivity index (SI) value of 133.0 for MAO-B, followed by EH6 at > 55.8. EH7 was a reversible and competitive inhibitor of MAO-B in kinetic and reversibility experiments with a Ki value of 0.034 ± 0.0067 µM. The molecular dynamics study documented that EH7 had a good binding affinity and motional movement within the active site with high stability. It was observed by MM-PBSA that the chirality had little effect on the overall binding of EH7 to MAO-B. Thus, EH7 can be employed for the development of lead molecules for the treatment of various neurodegenerative disorders.Halogens have been reported to play a major role in the inhibition of monoamine oxidase (MAO), relating to diverse cognitive functions of the central nervous system. Pyrazoline/halogenated pyrazolines were investigated for their inhibitory activities against human monoamine oxidase-A and -B. Halogen substitutions on the phenyl ring located at the fifth position of pyrazoline showed potent MAO-B inhibition. Compound 3-(4-ethoxyphenyl)-5-(4-fluorophenyl)-4,5-dihydro-1 L-DOPA, which is considered a safe therapy for PD . Ma. Ma11]. R and S forms [R and S forms from the racemic mixture of 2-pyrazoline is a considerable task in the identification of biologically more active isomers [Pyrazoline, or dihydropyrazole, is the partially reduced form of pyrazole, and 2-pyrazoline is the most stable tautomeric from in this class . The pre S forms . The rin S forms . The sep isomers . Many st isomers . At the  isomers ,33,34,35Regarding the role of halogens in MAO inhibition, it is generally known that FDA-approved MAO inhibitors like clorgyline, moclobemide, safinamide, and lazabemide are chlorine-containing drugs . The eviThe objective of the present study was focused on the in vitro evaluation of MAO-A and MAO-B inhibitions of pyrazoline/halogenated pyrazoline derivatives. The lead molecules were further studied for kinetics, reversibility, and blood–brain barrier (BBB) permeation. Detailed molecular dynamics established precisely the protein–ligand binding interactions of the halogenated molecules in their respective active sites of enzyme targets and the role of chirality towards MAO-B inhibition.EH sequence had potent inhibitory action against MAO-B, with residual activities of about 50% , and bromine-containing EH8 (IC50 = 0.69 µM). Comparing EH1 (IC50 = 5.38 µM) with EH7, the potencies for MAO-B greatly increased, i.e., by 85.4 times, based on their IC50 values, with the presence of –F instead of –H. Most of the compounds weakly inhibited MAO-A at 10 µM, with residual activities of >50%, except EH7 and EH8. Among them, EH7 showed the highest selectivity index (SI) value for MAO-B with 133.0, followed by EH6 with 55.8.At 10 µM, all derivatives in the bout 50% . Fluorinpara-position of the B ring of ethoxylated pyrazolines. Among the halogens, fluorine had the greatest impact on MAO-B inhibition with a sub-micromolar level range. It is worth noting that a highly electronegative fluorine atom-containing compound is two times more potent than marketed drugs for the inhibition of MAO-B, such as reversible-type lazabemide (IC50 = 0.11 µM) and irreversible pargyline (IC50 = 0.14 µM). Recent reports state that the anisotropy of charge circulation around the fluorine atom, which is the reason for the directional and stabilizing property of the active site, proves that halogen enhances the analogy of drugs of new molecules [The enzyme inhibition results obtained in the current study revealed that all of the halogenated pyrazoline derivatives showed good selective MAO-B inhibitory activity. The study mainly confirmed the effect of halogens on the olecules .50 values of 0.57, 0.053, and 2.01 µM, respectively [Recently, we evaluated MAO-B inhibitory activities for ethoxylated head of chalcone derivatives and found that chlorine-, fluorine-, or bromine-containing chalcones had good ICectively . CyclizaEH6 and EH7 were performed. Lineweaver–Burk plots and secondary plots showed that EH6 was a competitive inhibitor of MAO-A and MAO-B  to 89.5% . The recovery value was similar to that of the reversible reference lazabemide (from 35.3% to 75.7%); however, it was different from that of the irreversible inhibitor pargyline (from 41.3% to 44.9%), which was not recovered  . From th−6 cm/s . All halEH6, MAO-B_EH7, and MAO-B_EH8 attained equilibration at around 20 ns  can be used to estimate the conformational stability of a system during a production run . The difnd 20 ns . Howeverectively .EH1, EH6, EH7, and EH8 binding revealed that EH6 and EH7 had the highest active site-stabilizing effect on MAO-B, with overall average RMSD_AS values of 1.61 and 1.49 Å, respectively.Zeroing on structural influence of the ligands on the mechanistic behavior of the active site could provide more clues regarding their activity. The RMSD of the MAO-A and MAO-B active sites upon EH7 elicited some motional movement within the active site, which made it flexible enough to interact with crucial amino acid residues, it also exhibited good stability—just enough to be adequately anchored for overall binding  is a common molecular dynamic simulation analysis used in the estimation of the essential production run of a system when computed on a low-dimensional free energy space . This es systems A. This fnd MAO-A B.EH1, MAO-B_EH6, MAO-B_EH7, and MAO-B_EH8 were 0.88, 0.83, 0.68, and 0.75 Å, respectively  computes the residual deviation of a protein over a period of time using a reference position, such as the averaged residual position. RMSF estimates the level of fluctuation of residue from its original position . As depiectively C, while ectively D.The estimation of free binding energy using an MM/PBSA protocol can be employed to gain insight into the models and further obtain the semiquantitative values of their stability. We therefore used a representative snapshot and active site residue decomposition to further explore the time-wise bond interaction occurring between major residues in the active sites of MAO-A/MAO-B and the ligands. MM/PBSA has found useful application in the drug design space used in the estimation of binding affinity between ligands and biomolecules .EH7 had higher binding affinity  towards MAO-B than the other ligands, while the binding of EH1, EH6, and EH8 was recorded to be −4.11, −11.5, and −5.48 kcal/mol, respectively. Of note is the surprising high binding affinity  of EH7 for MAO-A. This could be due to the high electronegativity potential of fluorine, which is characteristic of compound EH7. In agreement with the IC50, the binding affinity of the compounds also followed the same trend.As indicated in EH7 was contributed by electrostatic interactions of Arg42, Arg87, Arg98, Lys296, and Lys386, with energy values of −28.4, −12.8, −15.3, −41.9, and −20.6 kcal/mol, respectively , EH6 (B), EH7 (C), and EH8 (D) with MAO-B is shown in The individual energy contributions of the active sites were determined using the MM/PBSA per residue energy decomposition protocol . The higectively C. In theectively B. Despitf Trp386 C. The liEH7 as a representative structure of the compounds, taking into consideration the fact that it has the best IC50 and total free energy values. We explored the differential binding and estimated the total free binding energy of the R isomeric form of EH7 (MAO-B_EH7R) and its S counterpart (MAO-B_EH7S) using MM/PBSA calculations. Our calculations revealed that MAO-B_EH7R and MAO-B_EH7S possess binding energies of −36.29 and −36.17 kcal/mol , which contribute van der Waals binding energies of −2.93, −1.58, and −1.00 kcal/mol, respectively  was refluxed with (0.0629 M) hydrazine hydrate under ethanol medium. After 14–16 h of reflux, the product was cooled, acidified with dil. HCl, washed thoroughly with water, filtered, dried, and recrystallized from ethanol. The progress of the formation of pyrazoline product was monitored by thin-layer chromatography . The synthetic route is depicted in EH1)3-(4-ethoxyphenyl)-5-phenyl-4,5-dihydro-1H-pyrazole  δ: 1.47–1.45 , 4.15–4.13 , 3.76 , 4.02 , 5.51 , 7.89–7.10 , 9.21. MS (ESI) m/z calcd. for C17H18N2O: 266.3376, found 266.3345.EH6)5-(4-chlorophenyl)-3-(4-ethoxyphenyl)-4,5-dihydro-1H-pyrazole  δ: 1.47–1.45 , 4.14–4.12 , 3.75 , 4.01 , 5.51 , 7.99–7.08  9.24 . MS (ESI) m/z calcd. for C17H17N2OCl: 284.3280, found 284.3276.EH7):3-(4-ethoxyphenyl)-5-(4-fluorophenyl)-4,5-dihydro-1H-pyrazole  δ: 1.47–1.45 , 4.15–4.13 , 3.72 , 4.04 , 5.51 , 7.96–7.03  9.23 . MS (ESI) m/z calcd. for C17H17N2OF: 266.3376, found 266.3345.EH8):5-(4-bromophenyl)-3-(4-ethoxyphenyl)-4,5-dihydro-1H-pyrazole  δ: 1.47–1.45 , 4.15–4.13 , 3.76 , 4.02 , 5.51 , 7.89–7.70 , 9.24. C17H17N2OBr: 345.2336, found 345.2340.m . The enzymes and chemicals were purchased from Sigma-Aldrich  [MAO activities were assayed as described previously, using recombinant human MAO-A and MAO-B and kynuramine (0.06 mM) and benzylamine (0.3 mM) as substrates, respectively; both substrate concentrations were 1.5 × KMO, USA) .50 values for MAO-B by compounds showing residual activities of <50% were determined. Kinetic studies were performed on the most potent inhibitors, i.e., EH6 and EH7 for MAO-A and MAO-B, at five concentrations of the substrates and three inhibitor concentrations, as previously described [The inhibitory activities of the four compounds against MAO-A and MAO-B were first observed at a concentration of 10 µM. ICescribed .EH7 were analyzed using a dialysis method after preincubating with MAO-B for 30 min, as previously described. The concentrations used were EH7 at 0.15 µM, lazabemide (a reversible MAO-B reference inhibitor) at 0.20 µM, and pargyline (an irreversible MAO-B reference inhibitor) at 0.30 µM. The relative activities for undialyzed (AU) and dialyzed (AD) samples were compared to determine the reversibility patterns [Reversibilities of patterns .EH1, EH6, EH7, and EH8; afterwards, geometrical and structural optimizations of these ligands were carried out using B3LYP/6-311 ++G  and the UFF forcefield of Gaussian 16, respectively [EH1, MAO-A:EH6, MAO-A:EH7, MAO-A:EH8, MAO-B:EH1, MAO-B:EH6, MAO-B:EH7, and MAO-B:EH8). The simulation process was carried out in accordance with an in-house protocol. The CPPTRAJ module of Amber18 software was used to analyze the trajectories emanating from the simulation process [The 3D structures of MAO-A (PDB ID: 2Z5X and MAO-B: 2V5Z), hereafter referred to as MAO-A and MAO-B, respectively, were retrieved from the Protein Data Bank (PDB) ,47. Moleectively . Active ectively . The sysectively . Eight s process ,54.EH7 were assessed at http://www.swisstargetprediction.ch (access on 23 May 2021) [The biological target and ADME predictions of the lead molecule of ay 2021) ,56.EH1, EH6, EH7, and EH8) was synthesized from their corresponding chalcones. All four compounds were explored for their MAO-A and MAO-B inhibitory activities. On the basis of previous studies regarding the role of halogens in various scaffolds with MAO inhibition activities, we mainly focused on the effect of various halogens on the pyrazoline core with the ethoxylated head. According to the results of this study, the synthesized halogenated pyrazolines exhibited good selective MAO-B inhibition. Fluorine-containing pyrazoline (EH7) exhibited an improved selectivity profile against the MAO-B isoform. Kinetics and reversibility studies revealed that EH7 is a selective and competitive inhibitor of MAO-B, with a recovery value similar to that of the reversible reference drug lazabemide. Pyrazolines are said to be optically active compounds, and, from the MM/PBSA values obtained, it can be clearly observed that chirality has minimal influence on the binding of the compound to MAO-B. It may be suggested that the absence of bulky groups on the N1 position of pyrazoline limited the role of chirality in the binding mode of the lead molecule in the current study.A series of four halogenated pyrazolines with ethoxylated heads (


Raphanus sativus L.) is a common Japanese ingredient, there are few reports of IgE-mediated immediate food allergy caused by Japanese radish.Although Japanese radish (A 48-year-old woman developed urticarial lesions on her hands after grating Japanese radish and also developed lip edema and oral itching when she ate a salad composed of raw Japanese radishes. Skin prick testing was positive to extract of grated Japanese radish. Moreover, immunoblotting analysis showed IgE reactivity in the patient’s serum to a single band at the 18 kDa in grated Japanese radish, suggesting that the heat-labile 18 kDa protein of raw Japanese radish may be a radish-specific antigen.To the best of our knowledge, this is the first case report of a patient with hand urticaria, lip angioedema, and oropharyngeal pruritus to raw Japanese radish through IgE-mediated immediate allergic reaction. Raphanus sativus L.), belonging to the mustard family (Cruciferae or Brassicaceae), is a common edible vegetable; the taproot of white radish is widely consumed in East Asia, both fresh and dried. Although mustard allergy is a well-known food hypersensitivity in children [Radish . Within a few minutes of grating raw JR, she developed urticaria on her hands at sites of contact. Although no symptoms developed when she ate boiled JR, after eating a salad containing raw JR she reported immediate onset oropharyngeal pruritus. Lip angioedema developed within minutes of ingestion. These manifestations improved spontaneously after 1 to 2 h. The laboratory test showed that the serum total immunoglobulin E (IgE) level was 138 IU/mL. Serum specific IgE antibodies against house dust mites and cedar pollens were positive (chemiluminescence enzyme immunoassay class 2). Although the result of the skin prick test (SPT) for extracts of boiled JR and normal saline were negative , a positive SPT was observed for the extract of raw JR and raw grated JR and histamine  . Equal amounts of the extracts (50 μg) were separated by SDS–polyacrylamide gel electrophoresis and blotted onto polyvinylidene difluoride membranes. After blocking, blots were incubated overnight with the patient’s serum or control. The serum of patients with Japanese cedar pollen allergy was used as control. The membranes were then incubated with appropriate alkaline phosphatase-conjugated goat-anti human IgE antibody , followed by detection with BCIP/NBT‐purple liquid substrate system for membranes . Immunoblotting analysis of non-denatured and denatured raw-grated radish showed IgE reactivity in the patient’s serum to a single band at 18 kDa and 2 bands at 18 and 65 kDa  while cooking and following a meal [Raphanus sativus L. var niger) [Sin a 1, Sin a 2, Sin a 3, and Sin a 4) from yellow mustard  [This is the first case of hand urticaria, lip angioedema, and oropharyngeal pruritus to raw JR through IgE-mediated immediate allergic reaction. Since 1974, 6 cases involving radish allergy have been reported in medical literature, besides the current case  , 4. A Jar niger) –6. Howevr niger) –13. The r niger) –13. Althalba L.) . HoweverBrassica nigra L.), and is known to have antimicrobial activity against bacteria and fungi [The mustard family vegetables, including JR, generate isothiocyanates, which are known as the pungent principle, through enzymatic hydrolysis of the corresponding glucosinolates . Allyl ind fungi –18. Althnd fungi . The punnd fungi , which iOral allergy syndrome (OAS) is defined as the symptoms of IgE-mediated immediate allergy localized in the oral and pharyngeal region and caused by contact with the acid and heat-labile antigen of raw fruits and vegetables –23. ApprIn conclusion, to the best of our knowledge, this is the first report of a patient with hand urticaria, lip angioedema, and oropharyngeal pruritus to raw JR through IgE-mediated immediate allergic reaction. Immunoblotting analysis suggests that the 18-kDa protein of raw JR may be a radish-specific antigen.


Currently, an increasing number of medical centers are treating mandibular head fractures surgically. Dedicated screws for compression osteosynthesis have been developed. However, due to the very limited size of the fractured bones and the considerable technical difficulties accompanying the execution of the fixation, there is little room for correction of the positioning and reinsertion of the screws. Therefore, knowing the optimal position of the fixation material is crucial for therapeutic success. The aim of this study is the evaluation of fixation screw position on the mandibular ramus height obtained in the treatment of the condylar head fracture. A total of 57 patients were included in this study. The loss of mandibular ramus height on computed tomography twelve months after mandibular head osteosynthesis was evaluated in relation to the initial distance of the screws from the fracture line, the angle of insertion of the screw into the bone, and the size of the protrusion to the inner side of the condyle. The relationship of the proximity of the screw to the fracture line, angulation, and the size of the protrusion with the loss of ramus height was confirmed. Conclusions: the optimal location for the superior screw is approx. 4 mm below the fracture line (with any angulation), inferior screw is approx. 8 mm (with any angulation), and anterior screw position is approx. 4–5 mm distant from fracture line with the best angulation of 130 degrees to the lateral mandible ramus surface in the coronal plane. Facial injuries are frequent trauma and most common in males 56.8–92.8%) ,2,3,4, a.8% ,2,3,Open treatment involves extracting a small fragment  of the mandibular head from the infratemporal fossa using raspators. The reduction of it is conducted with good visibility of the posterior surface of the mandibular head, followed by fixation with the larger fragment . This fixation can be performed with long screws of different constructions ,18,19,20The aim of this study is the evaluation of the fixation screw position on the mandibular ramus height obtained in treatment of the condylar head fracture.Patients were admitted as part of the in-patient emergency service, transferred from other hospitals, or the patients themselves reported to the outpatient clinic. After assessment of the general condition and possible respiratory and circulatory stabilization, treatment with maxillofacial surgery was planned. All patients had taken a multislice computed tomography (CT). Mandible head fracture classifications according to Neff were applied  19]..19].Inclusion criteria: recent trauma, condylar head fracture type B or C, fragment angulation > 10°, fragment overlapping > 2 mm, preauricular approach, and complete radiological documentation. Patients operated upon on Tuesdays and Thursdays received titanium fixation, while patients operated upon on Wednesdays and Fridays received magnesium fixation . Exclusion criteria: type A head fractures, fragment angulation ≤ 10°, fragment overlapping ≤ 2 mm, close  treatment, and failure of the patient to report for follow-up examinations.The clinical material consisted of 39 patients with single fractures of the mandibular head and 18 patients with bilateral fractures, making a total of 74 mandibular head fractures. Type B represented 19 fractures and type C represented 55 fractures. The study group was predominantly male (61) and the age of the patients ranged from 12 to 72 years (mean was 37 ± 18 years). There were 13 smoking patients and 22 internal medicine patients. Nicotinism was related to the existence of internal medicine diseases in patients.In the patients studied, there were 18 cases with dental fractures; 33 cases were accompanied by a mandibular body fracture, 12 cases were accompanied by a zygomatic bone fracture, and 13 cases had other distant injuries .Post-traumatic shortening of the mandibular ramus  of 8.1 ± 4.5 mm was noted. The deviation of the proximal  fragment from the axis of the mandibular condylar process was 37° ± 22°. The number of free mandibular head fragments  was 2.3 ± 1.1. Dislocation in the temporomandibular joint was observed in 62 cases and in the remaining 12 there was only displacement. Delay to treatment was 6.8 ± 8.8 day.Patients were operated in antibiotic prophylaxis under general anesthesia with intubation through the nose, floor of the mouth, or tracheostomy. In cases of concomitant mandibular fractures, mandibular head fractures were fixated first from the preauricular approach. Efforts were made to insert 3 screws each time for the osteosynthesis of one mandibular head. Headless and solid compression screws made of titanium alloy and magnesium alloy  were used. The lengths of the screws used were 14 mm, 16 mm, or 18 mm and the system was: 1.8 or 2.2 .An attempt was made to fix the fracture with three screws: the first in the superior position (S), the second in the inferior position (I), and the third in the anterior position (A) in relation to the other two . All scrNr is the number of series of pixels with density level i and length k; Ng—number of levels for image optical density; p is probability, log is common logarithm [Nr—number of pixels in series; [The structure of the bone union was also examined by means of digital texture analysis on coronary sections of the mandibular head . ROI covogarithm ; Nr—numb series; ,27. The  series; , which r series; ,23.t-test) or median (W-test) comparison, χ2 tests of independence, analysis of regression, and one-way analysis of variance. Detected relationships were assumed to be statistically significant when p < 0.05. Statgraphics Centurion version 18.1.12  was used for statistical analyses.The statistical analysis includes the feature distribution evaluation, mean . A similar relationship exists for the inferior screw  and the anterior screw , as shown in The mean values of the screw distance from the fracture line, the screw insertion angle in relation to the lateral plane of the mandibular ramus, and the extent of penetration to the internal surface of the condylar process obtained for the study group are shown in 2 = 51%; p < 0.05). In this study, this angle varied from 96.5 degrees to 130 degrees . Such a weak relationship was observed for the inferior screw protrusion . For anterior screw incidences of protrusion, the relation was moderately strong . The collective results are presented in A total of 21 cases of superior screw protrusion were found in this study; the range of the protrusion was from 1 mm to 5.8 mm. A total of 29 cases of medial protrusion were noted for the inferior screw position with a range of 1.00–8.32 mm. Only five cases of medial protrusion were found for the anterior screw with a range of 2.25–2.50 mm. A relatively weak relationship between the superior screw protrusion and ramus height loss was found . The value of the correlation coefficient indicates a relatively weak relationship between the variables.When examining the quality of the union, it is important to note that the optical density at the consolidation site is higher than that of the surrounding bone 12 months after surgery. In addition, structural analysis of the image of this bone at the post-fracture site (BI), indicates a bone different from the cancellous bone . Bone de2 = 8%; p < 0.05).Only the distance of superior screws from the fracture line is related to BI: the closer to the fracture line of the inserted screw, the closer the bone structure in the union is to the cancellous bone  and anterior screw , whose inclination is associated with BI values . The other two screw locations do not show a relationship of screw tip protrusion size with the Bone Index.The amount of superior screw protrusion to the medial side of the mandible is directly proportionally weakly related to an increase in BI values . The case is similar for the inferior screw , where the distance was 9.42 ± 3.52 mm. The quality of consolidation as measured by the BI value in type B fractures was 0.62 ± 0.28. The study also noted that larger insertion angles of the superior screw were associated with an increasing loss of ramus height. This relationship is moderately strong . The amount of ramus height loss was the same in type C fractures (3.93 ± 3.09 mm) than type B (4.07 ± 2.61 mm).Assessing only type B fractures, a superior screw distance of 6.30 ± 2.83 mm from the fracture line and its moderately strong association with the loss of ramus height was noted  and a moderately strong inverse proportional association of anterior screw distance from the fracture line (5.46 ± 0.89 mm) with the amount of ramus height loss . A moderately strong inverse relationship was observed for the size of the anterior screw insertion angle (118° ± 12°) and the amount of ramus height loss . On the other hand, in type C fractures (as opposed to the absence of such a relationship in type B), an association of the protrusion of the screw to the inner side was noted. For protrusion in the superior screw (1.21 ± 1.96 mm), a weak inverse relationship with ramus height loss was noted . For the inferior screw, a relatively weak direct relationship to ramus loss was also found . A small number of fixing material protrusion was not allowed for the statistical analysis of the anterior screw protrusion with ramus loss.As far as only type C fractures were considered, BI was similar (0.67 ± 0.29), as with type B fractures. It was noted that there was a weak direct proportional association of the superior screw distance (3.20 ± 1.15 mm) with the amount of ramus height loss . The primary post-traumatic overlap of the fragments (ramus shortening) was directly proportionally associated with the final treatment outcome . However, this relationship was relatively weak.Between one and four screws per osteosynthesis were used in the study group of patients. However, two screws or three pieces were most commonly used. Combining these subgroups separated patients with fixations with a smaller number of screws (one or two used) accounting for 61 fixations and a group treated with a larger number of screws (three or four screws) accounting for 13 osteosyntheses. The scanty-screw (3.81 ± 2.81 mm) vs. multi-screw (4.69 ± 3.61 mm) subgroups did not differ in the amount of ramus height loss or in the Bone Index, i.e., union quality . A moderately strong direct proportional relationship was detected between the post-traumatic angulation of bone fragments and the final resultant loss of mandibular ramus height  in relation to the fracture line leads to a large loss of ramus height. It can be assumed that the mechanical fixation of the screw in the distal fragment  is too weak and that masticatory forces can cause a gradual downward tilt of the medial pole of the mandibular head. On the other hand, an inverse relationship can be observed with regard to the influence of the location of the interior screw. The closer the inferior screw is to the fracture line, the better the long-term outcome (less loss of ramus height). The regression lines intersect at a point corresponding to a distance of approximately 6.5 mm from the fracture line. The superior and inferior screw cannot be inserted in the same place. The observation of the two regression lines reveals a flattening of the regression lines, which allows indicating the optimal distance for each screw. For the superior screw, the optimal insertion site is about 4 mm below the fracture line, while for the inferior screw, the optimal insertion site is 7–8 mm below the fracture line. A guideline from these studies for the location of the anterior screw is that it is best placed 4–5 mm below the fracture line. Sometimes only two screws are possible to place . It woulIn light of these studies, the role of the anterior screw seems important in the osteosynthesis of the mandibular head. If there is room for maneuver, better long-term anatomical results will be obtained when the screw insertion angle is close to 130 deg. (i.e high) in the coronal plane (and worse results when it is closer to 96 deg.). As far as the angulation of the other screws is concerned, the operator is free to act and can be guided by the assessment of the intraoperative situation in choosing the insertion angle of the superior and inferior screws. All angles from the range 90°–140° can be utilized without any negative influence on later mandible ramus height.p < 0.05). However, there is a tendency that the closer the superior screws are, the higher is the BI union. If one assumes that the appearance of the bone at the fracture site is different for years than the intact bone [The relationship between the amount of protrusion of the superior screw and the amount of loss of the mandibular ramus height is opposite to that of the inferior and anterior screws. The amount of protrusion of the latter two screws on the inner side of the mandibular condylar process is directly proportionally related to the amount of ramus height loss after 12 months . This seems easy to explain. The more the screw protrudes on the medial side of the mandible , the more it damages, e.g., presses on tissues that supply blood to the mandibular head, i.e., the lateral pterygoid muscle . Surprisact bone , one couThus, it seems that not only the screw diameter , length A weakness of the study is the difficulty in assessing the role of the anterior screw due to the few patients fixated by three screws (this is a requirement for the existence of a screw in the anterior position). Despite the inclusion of 57 patients and 74 fractures, this material needs to be enlarged and especially continued in a multicenter dimension.Based on the collected clinical material and the analysis of the mandibular ramus height loss at 12 months after surgery, it can be noted that the optimal location for the superior screw after the reduction of a mandibular head fracture appears to be approx. 4 mm below the fracture line (with any angulation), together with the inferior screw approx. 8 mm (with any angulation), and anterior screw position approx. 4–5 mm distant from fracture line, with the best angulation of 130 degrees in the coronal plane. The use of wrong angles and too long screws lead to the protrusion of the screw tip on the mesial/anterior side of the mandible head and may worsen the long-term results of the treatment of head fractures.


Signalling of the epithelial splicing variant of fibroblast growth factor receptor 2 (FGFR2b) triggers both differentiation and autophagy, while the aberrant expression of the mesenchymal FGFR2c isoform in epithelial cells induces impaired differentiation, inhibition of autophagy as well as the induction of the epithelial‐mesenchymal transition (EMT). In light of the widely proposed negative loop linking autophagy and EMT in the early steps of carcinogenesis, here we investigated the possible involvement of FGFR2c aberrant expression and signalling in orchestrating this crosstalk in human keratinocytes. Biochemical, molecular, quantitative immunofluorescence analysis and in vitro invasion assays, coupled to the use of specific substrate inhibitors and transient or stable silencing approaches, showed that AKT/MTOR and PKCε are the two hub signalling pathways, downstream FGFR2c, intersecting with each other in the control of both the inhibition of autophagy and the induction of EMT and invasive behaviour. These results indicate that the expression of FGFR2c, possibly resulting from FGFR2 isoform switch, could represent a key upstream event responsible for the establishment of a negative interplay between autophagy and EMT, which contributes to the assessment of a pathological oncogenic profile in epithelial cells. For MTOR and PKCε silencing, clones were transiently transfected with MTOR small interfering RNA (MTOR siRNA) , or stably transfected with PKCε Plasmide shRNA (h) vector , or an unrelated siRNA/shRNA as a control, using Lipofectamine 2000 transfection reagent . For growth factor stimulation, cells were left untreated or incubated with FGF2  100 ng/mL for 24 hours at 37°C. For inhibition of FGFR2c tyrosine kinase activity, cells were pre‐incubated with a specific FGFR2 tyrosine kinase inhibitor, SU5402 25 µmol/L  for 1 hour before treatments with growth factors (GFs). To inhibit AKT or MTOR, cells were incubated with AKT‐specific inhibitor AKT‐I‐1/2  or with the specific MTOR inhibitor rapamycin , respectively, for 1 hour at 37°C before being treated with FGF2 in the presence of each inhibitor.2.2t test, and significance level has been defined as P > .05.Migration assay was performed using 24‐well transwell migration Boyden chambers  pre‐coated with matrigel  as reported.2.3t test was performed, and significance levels have been defined as P < .05.HaCaT clones, grown on coverslips, were processed as previously reported.2.4t test was performed, and significance levels have been defined as P < .05.Cells were lysed, and total proteins were collected, separated and blotted as reported.2.5Oligonucleotide primers necessary for target genes and the housekeeping gene were chosen by using the online tool Primer‐BLAST 2.6RNA was extracted and retrotranscribed using the iScriptTM cDNA synthesis kit  as previously reported.2.7t test was performed, with significance levels defined as p values < 0.05. *Real‐time RT‐PCR was performed using the iCycler real‐time detection system (iQ5 Bio‐Rad) with optimized PCR conditions as reported.33.1Since MTOR is the main pathway involved in the inhibition of autophagy induced by aberrant FGFR2c expression in human keratinocytes,We then investigated the impact of MTOR inhibition on the invasion ability, displayed by pBp‐FGFR2c clones,In order to further assess the outcome of forced reactivation of MTOR‐dependent autophagy on FGFR2c‐mediated EMT in epithelial context, MTOR protein depletion was carried out in HaCaT clones by specific siRNA transfection. Western blot analysis showed that, similarly to what observed in the presence of rapamycin, in HaCaT pBp‐FGFR2c cells stimulated with FGF2, MTOR silencing was not only sufficient to reverse the decrease of LC3‐II in an increase and to block SQSTM1 accumulation Figure , confirmSince AKT is the substrate acting upstream MTOR in FGFR2c‐mediated inhibition of autophagy,Since FGFR2c‐triggered EMT is driven by PKCε‐dependent induction of Snail1, STAT3 and FRA1,3.2The possibility that FGFR2c could orchestrate a fine interplay between autophagy and EMT in epithelial context is also sustained by the evidence that protein kinase C isozymes, including PKCε, are also key regulators of the autophagic pathway.As last aim, to confirm the central role of FGFR2c in regulating all the observed, intersected effects between autophagy and EMT, we used the FGFR2 tyrosine kinase inhibitor SU5402. In HaCaT pBp‐FGFR2c clones, the presence of SU5402 was sufficient to abolish all the responses to FGF2, not only in terms of FGFR2c, PKCε and MTOR/S6K phosphorylation Figure , as expeOverall, our results represent the first indication that, at least in the context of human keratinocytes, the aberrant expression of FGFR2c, usually stemming from altered FGFR2 isoform switch, could be the upstream event leading to the activation of oncogenic signalling pathways intersecting with each other and cooperating in the establishment of the negative loop between EMT and autophagy, which contributes to the early steps of tumour development.The authors declare that they have no competing interests.Danilo Ranieri: Conceptualization ; Investigation (lead); Writing‐original draft (lead). Monica Nanni: Data curation ; Investigation . Luisa Guttieri: Investigation . Maria Rosaria Torrisi: Conceptualization ; Writing‐review & editing . Francesca Belleudi: Conceptualization ; Project administration (lead); Writing‐review & editing (lead).Fig S1Click here for additional data file.Fig S2Click here for additional data file.


Yersinia pestis substrate-binding protein YfeA contains two polyspecific metal-binding sites, and site 2 is capable of collaborating with other protein molecules for inter-protein metal coordination. The inter-protein metal coordination can occur by multiple coordination geometries and may be flexible enough to enable metal transfer to multiple metal-binding proteins.The  Yersinia pestis, the causative microbe of plague. YfeA folds into a monomeric c-clamp like other substrate-binding proteins and has two metal-binding sites (sites 1 and 2). Site 2 is a bidentate surface site capable of binding Zn and Mn atoms and is a unique feature of YfeA. Occasionally, the site 2 residues of two YfeA molecules will cooperate with the histidine tag of a third YfeA molecule in coordinating the same metal and lead to metal-dependent crystallographic packing. Here, three crystal structures of YfeA are presented at 1.85, 2.05 and 2.25 Å resolution. A comparison of the structures reveals that the metal can be displaced at five different locations ranging from ∼4 to ∼16 Å away from the canonical site 2. These observations reveal different configurations of site 2 that enable cooperative metal binding and demonstrate how site 2 is dynamic and freely available for inter-protein metal coordination.The substrate-binding protein YfeA  is a polyspecific metal-binding protein that is crucial for nutrient acquisition and virulence in  Fractions containing the eluent were pooled and loaded onto a HiTrap Q HP column , washed and eluted using a linear NaCl gradient from 0.0 to 1.0 M NaCl. Fractions containing the eluent were pooled and further purified by gel filtration on a HiLoad 26/600 Superdex 200 pg column . The final purified protein was concentrated to 18 ± 5 mg ml−1 in 20 mM bis-Tris propane pH 6.3, 50 mM NaCl, 0.05%(w/v) NaN3 for crystallization.Recombinant YfeA-His2.2.2 triggered YfeA oligomerization, 5 mg ml−1 YfeA was incubated with and without 10 mM MnCl2 in 20 mM bis-Tris propane pH 6.3, 50 mM NaCl, 0.05%(w/v) NaN3 for 2.5 h at room temperature. The mixture was analyzed by gel filtration using a Superdex 200 10/300 GL column .To determine whether MnCl2.3.10 were grown by the hanging-drop vapor-diffusion method at 20°C using a 1:1 ratio of 32%(w/v) PEG 4000, 20 mM bis-Tris propane pH 6.3, 50 mM NaCl, 0.05%(w/v) NaN3, 10 mM MnCl2 and 18 ± 5 mg ml−1 YfeA-His10. X-ray diffraction data were collected at the X-ray absorption-edge energy (K edge) for zinc or iron as described previously  was used as a search model  Superpose in Coot , whereas generally the YfeA crystals had one YfeA molecule in the asymmetric unit and belonged to an orthorhombic space group with unit-cell dimensions a = 41, b = 52, c = 113 Å or a = 62, b = 66, c = 66 Å . Structural alignment of each YfeA protomer from PDB entries 5uyg and 5uxs  yielded an r.m.s.d. of 0.54 ± 0.09 Å for 269–272 pairs of matched Cα atoms, indicating that the YfeA protomers here are highly similar to YfeA monomers. The r.m.s.d. among the present YfeA protomers is 0.47 ± 0.16 Å for 271–281 pairs of matched Cα atoms, indicating that the YfeA protomers are highly similar to one another. Some protomer pairs had more matched Cα atoms than others due to the resolution of additional residues in the carboxy-terminal His10 tag in some protomers. YfeA can also bind ancillary metals at additional surface sites . These metals are coordinated by multiple protomers and enable crystal contacts in the lattice, but have incomplete coordination spheres due to missing solvent molecules or other nonprotein ligands . Three histidine residues of the His10 tag are resolved in one protomer and no histidine residues are resolved in the other protomer in the asymmetric unit. The arrangement of ligands results in square bipyramidal coordination geometry, where the His314 ligand is at the apex of the pyramid and the other four ligands form the square base. Square pyramidal coordination geometry is commonly observed in protein–manganese . Therefore, two site 2 motifs participate in metal coordination. Five histidine residues of the His10 tag are resolved in one protomer and no histidine residues are resolved in the other protomer in the asymmetric unit. The four ligands form bond angles of ∼110–115° in this arrangement, which is consistent with tetrahedral coordination geometry. Tetrahedral coordination geometry is widespread in protein–transition metal complexes and is used at site 1 to coordinate zinc, manganese and iron  Yfe is required for iron transport and full virulence of plague infection  thc). Similarly, five histidine residues of the His10 tag are resolved in one protomer and no histidine residues are resolved in the other protomer in the asymmetric unit. There are no ancillary metals present in the structure.In YfeA site 2 inter-coordinated structure 3, the metal is coordinated by the same ligands as in structure 2 to achieve tetrahedral coordination geometry Fig. 2c. SimilM MnCl2 for 2.5 h and the mixtures were seperated by gel filtration to determine whether MnCl2 triggered YfeA oligomerization in solution. The elution position under both conditions is consistent with monomeric YfeA, indicating that the perceived oligomerization is a property of crystallization  in an internal translocation relay before iron is incorporated into a growing mineral for nutrient storage  and Y. ruckeri (GenBank accession AJI94945.1) were found to contain a site 2 lysine substitution for glutamate or a lysine insertion between the glutamate and histidine, but the histidine is conserved in all sequences . This metal-binding behavior is consistent with the characterization of YfeA as a zinc sink  or multiple site 2 motifs . Only one site 2 motif is needed to bind the metal, and the metal can be coordinated by backbone or side-chain ligands, making a potential metal handoff between proteins feasible. Participation of the YfeA His10 tag in metal coordination is intriguing because the zinc-binding SBP ZnuA has a histidine-rich loop near ZnuA site 1 that has been implicated in metal transfer to/from site 1  and global (for example buried versus surface) environments that impact metal binding and possibly function. The c-clamp fold contributes all of the coordinate interactions at site 1, limiting metal binding to a single configuration in the buried site. On the other hand, protein and bulk solvent combine to achieve all of the coordinate interactions for the site 2 surface site. Nine YfeA ancillary metal-binding sites have previously been identified . Therefore, His-tag zinc coordination shows that (i) the site 2 metal site is sufficiently close to the surface to participate in intermolecular metal coordination and (ii) the His-tag histidine is likely to mimic the metal coordination geometry that could occur with an as-yet-unidentified biological binding partner.4fi3; Korkhov et al., 2012et al., 1998The possibility of metal transfer from YfeA site 2 to the Yfe transporter was considered and an alignment of YfeA against BtuF in complex with the Btu transporter (PDB entry YfeA oligomer crystal 1, form 1, 7me1PDB reference: crystal 2, form 2, 7me2PDB reference: crystal 3, form 2, 7me3PDB reference:


Rab proteins are small GTPases that act as molecular switches for intracellular vesicle trafficking. Although their function is mainly regulated by regulatory proteins such as GTPase-activating proteins and guanine nucleotide exchange factors, recent studies have shown that some Rab proteins are physiologically phosphorylated in the switch II region by Rab kinases. As the switch II region of Rab proteins undergoes a conformational change depending on the bound nucleotide, it plays an essential role in their function as a ‘switch’. Initially, the phosphorylation of Rab proteins in the switch II region was shown to inhibit the association with regulatory proteins. However, recent studies suggest that it also regulates the binding of Rab proteins to effector proteins, determining which pathways to regulate. These findings suggest that the regulation of the Rab function may be more dynamically regulated by phosphorylation than just through the association with regulatory proteins. In this review, we summarize the recent findings and discuss the physiological and pathological roles of Rab phosphorylation. The Rab family is a member of the Ras small GTPase superfamily, which consists of more than 60 proteins in humans (reviewed in ). Rab faWhen a Rab protein is biosynthesized, it is first recognized by Rab escort proteins (REPs) in the cytosol and then recruited to the Rab geranylgeranyl transferase (RabGGTase) complex, by which it undergoes geranylgeranylation modification ,3 FigurA. GeranyOnce inserted into the lipid bilayer, each Rab controls different vesicular trafficking pathways depending on its localization. The localization of a Rab protein is thought to be defined by its activator, a guanine nucleotide exchange factor (GEF). GEF activity accelerates the dissociation of GDP from the Rab protein, and this then induces the binding of cytoplasmically abundant GTP to the Rab protein (reviewed in ). The GTThe destination of vesicular trafficking is defined by multiple factors. One factor is the combination of a soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) protein present on the vesicle membrane (v-SNARE) and multiple SNARE proteins on the target membrane (t-SNARE) that promotes the fusion of the membranes (reviewed in ). AnotheAlthough it has been textbook knowledge that Rab is regulated by GEFs, GAPs and GDIs, as outlined above, recent studies have revealed that the Rab function is also regulated by phosphorylation. The phosphorylation of Rab proteins has been shown not only to inhibit their interaction with GEFs and GDIs, but also to let them acquire different effector proteins, suggesting the existence of alternative trafficking routes activated by Rab phosphorylation B,C. In tLeucine-rich repeat kinase 2 (LRRK2) is one of the responsible gene products for familial Parkinson’s disease (PD) ,10. PD iLRRK2 is a large protein kinase consisting of 2527 amino acids, harboring a GTP-binding domain called a Ras-of-complex proteins (ROC) domain followed by a carboxyl-terminal of the ROC (COR) domain, an auxiliary domain of the ROC domain and a protein kinase domain  Figure . PathogeIn 2016, Steger and colleagues discovered that several Rab proteins are phosphorylated by LRRK2 under physiological conditions . They idSteger and colleagues further showed that LRRK2 phosphorylates fourteen Rab proteins, namely Rab3A/B/C/D, Rab5A/B/C, Rab8A/B, Rab10, Rab12, Rab29, Rab35 and Rab43, upon co-overexpression in HEK293 cells  Figure . Among tIn a paper published in 2019 by Berndsen and colleagues, the endogenous phosphorylation of Rab10 and Rab12 in R1441C LRRK2 knock-in MEFs was shown by immunoblotting using phospho-specific antibodies . In thisA similar result was reported by other groups. Jeong and colleagues showed the phosphorylation of a panel of Rab proteins by recombinant wild-type (WT) LRRK2 in an in vitro assay using isotope-labelled ATP . They foBased on these results, it is now widely accepted that LRRK2 is a Rab kinase, and its physiological and pathological relevance has attracted much attention.Since the discovery of Rab phosphorylation by LRRK2, researchers have been focusing on detecting Rab phosphorylation in human body fluid samples as a potential method for diagnosis. Thirstrup and colleagues employed a combined proteomics and phosphoproteomics analysis on human peripheral mononuclear cells (PBMCs) and identified the LRRK2-dependent phosphorylation of Rab10 and Rab12 at Thr73 and Ser106, respectively . As PBMCIn a more recent report by Fan and colleagues, the authors compared various types of blood cells and found, by immunoblotting, that neutrophils are superior to PBMCs in the detection of Rab10 phosphorylation . As neutAnother report by Karayel and colleagues failed to find a difference in the levels of Rab10 phosphorylation in neutrophil samples between healthy controls and idiopathic PD patients, either using immunoblotting or using mass spectrometry . HoweverGiven that neurodegeneration occurs in PD, it is important to investigate whether Rab10 phosphorylation can be detected in the brain and whether there is a difference between the controls and the PD patients. Di Maio and colleagues examined the Rab10 phosphorylation in postmortem brains using immunohistochemistry . They deThe reason for this discrepancy between reports is not clear, but it may be due to the varying levels of Rab10 phosphorylation among individuals. Therefore, further studies with a larger number of subjects and different cohorts are required to clarify the usefulness of Rab10 phosphorylation in diagnosing PD using blood samples, as well as to examine the change in Rab10 phosphorylation in PD patient brains.Rab proteins are phosphorylated by LRRK2 via the highly conserved switch II region, which regulates the hydrolysis of GTP and coordinates the binding to various regulatory proteins . TherefoThe phosphorylation of Rab8 and Rab10 by LRRK2 is reported to play an important role in lysosomal homeostasis. The treatment of cells with lysosomotropic agents ) results in lysosomal overload, in which lysosomes become enlarged. In mouse macrophagic Raw264.7 cells, in response to the overload, lysosomal contents are actively released to the exterior of the cells. Intriguingly, Eguchi and colleagues have shown that the phosphorylation of Rab8 and Rab10 by LRRK2 is required for this release, as the treatment of cells with LRRK2 inhibitors or knockdown of Rab8 or Rab10 caused an enlargement of lysosomes and a concomitant decrease in the amount of extracellularly released Cathepsin D, a lysosomal enzyme ,36,37. TIn macrophages, the activation of LRRK2 is also observed upon endomembrane damage caused by an infection of pathogens or lysosomotropic drug treatment . EndomemThere is also a study reporting that LRRK2 phosphorylates Rab10 on early immature macropinosomes in macrophages . Rab10 oSeveral studies have reported the potential role that the LRRK2-Rab pathway plays in blocking the formation of primary cilia. A prerequisite for primary cilia formation is the dissociation of centrosomal protein of 110 kDa (CP110) from the mother centriole, which becomes the basal body in ciliogenesis. Tau-tubulin kinase 2 (TTBK2) is a kinase required for this dissociation, and it is recruited to the pericentriolar region through the interaction with centrosomal protein 164 (CEP164), a centriole protein. It was shown that this recruitment of TTBK2 to the pericentriolar region is inhibited by the hyperphosphorylation of Rab10  Figure D. In addThis link between LRRK2 and ciliogenesis may also affect the sonic hedgehog (Shh) signaling. The Shh signaling requires the formation of primary cilia, to which the downstream transcription factors, such as Gli1 and Gli2, are localized ,46. LRRKThe effects of Rab phosphorylation on its interaction with its effector have also been investigated in the context of structural biology. In 2020, Waschbüsch and colleagues reported the structure of phosphorylated Rab8A (pRab8A) in complex with the RH2 domain of RILPL2 using X-ray crystallography . RILPL2 In PD, the aggregation of α-synuclein into inclusion bodies called Lewy bodies is a prominent pathological feature ,14. It hA number of studies have looked into the effects of LRRK2 mutations on the accumulation of α-synuclein. In these reports, dopaminergic neurons differentiated from induced pluripotent stem cells (iPSCs) prepared from PD patients harboring the G2019S LRRK2 mutation consistently showed increased amounts of cytoplasmic α-synuclein, presumably due to posttranslational alterations ,54,55,56Numerous studies have also looked into the roles that LRRK2 and Rab may play in α-synuclein propagation. However, there has been conflicting evidence, and it remains controversial whether or not LRRK2 is involved in α-synuclein pathology.In 2018, Henderson and colleagues reported that LRRK2 activity only has minor effects on α-synuclein pathology . PrimaryIn a paper published in 2019, the same group further reported that in non-transgenic mice, LRRK2 inhibition does not lead to increased protection from α-synuclein pathology or neuron death . They maLrrk2-targeting guide RNAs in primary neurons cultured from Cas9 transgenic mice. Conversely, in primary neurons from transgenic mice expressing G2019S LRRK2, an increase in α-synuclein aggregation was observed. Furthermore, LRRK2 G2019S transgenic mice exhibited an acceleration of α-synuclein aggregation and the degeneration of dopaminergic neurons in the SNpc increased degeneration-associated neuroinflammation and behavioral deficits. This was further validated in a human context by establishing a human α-synuclein transmission model using iPSC-derived neurons (iNs). In the iNs derived from the PD subjects, the G2019S mutation led to the enhanced aggregation of α-synuclein, while the loss of LRRK2 decreased it. Altogether, the authors suggested a strong link between the LRRK2 activity and α-synuclein propagation.On the other hand, Bieri and colleagues published a paper in 2019 reporting that LRRK2 does modify α-synuclein pathology and spread in mouse models and human neurons . They peCaenorhabditis elegans exhibited reduced levels of α-synuclein aggregation compared to the wild-type, and this could be rescued with the ectopic expression of Rab35, but not with the T72A mutant of Rab35, which cannot be phosphorylated by LRRK2. Furthermore, the reduced α-synuclein propagation phenotype in lrk-1 mutant C. elegans could be overridden with the expression of the constitutively activated Q67L mutant of RAB35, accompanied with worsened degenerative phenotypes including nerve degeneration and life span. Lastly, using an α-synuclein transgenic mouse model, the authors showed that the administration of an LRRK2 kinase inhibitor led to a decrease in α-synuclein aggregation, accompanied by an increasing interaction of α-synuclein with the lysosomal enzyme Cathepsin D, suggesting an enhanced association with the lysosomal degradation pathway. This prompted the authors to suggest that the LRRK2-mediated RAB35 phosphorylation positively regulates α-synuclein propagation.In 2018, Bae and colleagues also reported that LRRK2 plays a role in α-synuclein propagation, proposing a more detailed mechanism in which one of the substrates of LRRK2, Rab35, participates in α-synuclein propagation . They shCollectively, although the above studies examining the expression levels of α-synuclein in iPSC-derived dopaminergic neurons appear to conclude that G2019S LRRK2 exacerbates α-synuclein accumulation, it remains controversial whether LRRK2 plays a role in the propagation of α-synuclein pathology, especially in an in vivo setting. The reason for the discrepancies is unclear at the moment, but it may be due to multiple factors including differences in the methods for the preparation of α-synuclein seeds  has been implicated in PD pathogenesis as several genome-wide association studies on sporadic PD found that a genomic locus containing Rab29 is associated with an increased risk of developing PD . IntriguFirstly, Rab29 is a physiological substrate of LRRK2. LRRK2 phosphorylates Rab29 at Thr71 or Ser72, or both, in the switch II region ,24,63. TSecondly, the overexpression of Rab29 in cultured cells activates LRRK2. When Rab29 was co-overexpressed in HEK293 cells with WT LRRK2, the autophosphorylation of LRRK2 at Ser1292 as well as Rab10 phosphorylation was dramatically increased . This acThe dependence of LRRK2 activity on Rab29 was also supported by a paper by Eguchi and colleagues . As mentRab29 in cells or mice has no impact on the phosphorylation of LRRK2 substrates. In a paper by Kalogeropulou and colleagues, they generated Rab29 KO mice and found no effect on the phosphorylation of LRRK2 substrates in vivo [However, the mechanism of how Rab29 activates LRRK2 remains unclear. Several reports have shown that the knockout (KO) of  in vivo . Moreove in vivo .Collectively, it requires further investigation to elucidate the connection between Rab29 and LRRK2, especially in the context of PD pathogenesis.Phosphatase and tensin homolog deleted from chromosome 10 (PTEN)-induced kinase 1 (PINK1) is a responsible gene for a type of autosomal recessive juvenile parkinsonism . The loss-of-function mutations in PINK1 have been found in PARK6 patients. It has been well documented that PINK1 functions as a sensor of mitochondrial depolarization and facilitates the degradation of damaged mitochondria by recruiting and phosphorylating Parkin, another gene responsible for AR-JP (PARK2), as well as ubiquitin, leading to the degradation of the damaged mitochondria via autophagy  (reviewed in ). TherefLai and colleagues found that the phosphorylation of Rab1B, 8A, 8B and 13 is regulated by PINK1 . These RAs Rab8A Ser111 is within the RabSF3 motif in close proximity to the switch II region containing Thr72, phosphorylation on either site may influence the efficiency of the other phosphorylation. Indeed, Rab8A harboring phospho-Ser111 seemed to be less efficiently phosphorylated by LRRK2 at Thr72 than WT Rab8A in vitro as well as in cultured cells . These rLeucine-rich repeat kinase 1 (LRRK1) is a paralog of LRRK2. These proteins share a similar domain architecture including ankyrin repeats, leucine-rich repeats, ROC, COR, a protein kinase domain and carboxyl-terminal WD40 repeats A,B. LRRKIt has been reported that LRRK1 regulates the mitotic spindle orientation by phosphorylating cyclin-dependent kinase 5 regulatory subunit-associated protein 2 (CDKRAP2), a centrosomal protein that plays a role in the maturation of centrosomes. The interaction of CDK5RAP2 with γ-tubulin was promoted upon phosphorylation by LRRK1, leading to an increase in the microtubule (MT) formation activity. When LRRK1 was knocked down, astral MTs were markedly decreased, and the MT nucleation activity of the centrosomes was downregulated, eventually leading to a loss of spindle orientation control .Glued/dynactin-1 to the MT plus ends. Additionally, activated EGFR is known to migrate on early endosomes along MTs as the endosomes undergo maturation. It was reported that the LRRK1-dependent phosphorylation of CLIP-170 enhances the interaction with p150Glued/dynactin-1, thereby facilitating the transport of EGFR [LRRK1 was also shown to phosphorylate cytoplasmic linker protein 170 , a protein involved in dynein-mediated transport, thereby playing a role in EGFR trafficking . CLIP-17 of EGFR .Importantly, Hanafusa and colleagues recently provided evidence that shows that GTP-bound active-state Rab7A is also phosphorylated by LRRK1 at the endosomal membrane . Rab7A iMalik and colleagues recently reported that LRRK1 and LRRK2 have distinct substrate spectra . LRRK1 aMalik and colleagues also found that the phosphorylation of Ser72 in Rab7A was most significantly affected when LRRK1 was knocked out in MEFs, which suggests that LRRK1 is the major kinase for the phosphorylation of Ser72 in Rab7A, at least in MEFs. Of note, the authors showed that the phosphorylation of Rab7A did not change its interaction with RILP. Therefore, whether the effector protein of phosphorylated Rab7A plays a role in EGFR signaling remains controversial. Meanwhile, the authors also showed that stimulation of MEFs with phorbol esters, which activates protein kinase C (PKC), increased LRRK1-mediated Rab7A phosphorylation, and that this was inhibited by the treatment with Gö6983, a PKC inhibitor. Collectively, these reports imply that LRRK1 may be involved in the EGFR signaling pathway downstream of PKC via its kinase activity A.Another study employing a system-wide spatiotemporal characterization of the ErbB-family receptor signaling complex identified LRRK1 in the EGFR receptor signaling complex as well as the cytosolic receptor-free subcomplex .TBK1 (TANK-binding kinase 1) had been known to facilitate efficient mitophagy by phosphorylating optineurin (OPTN), Nuclear domain 10 protein 52 (NDP52)/Calcoco2 and Sequestosome-1 (SQSTM1), which are adaptors that link the ubiquitin chain formed on the mitochondrial outer membrane to the autophagy machinery. The TBK1-dependent phosphorylation of OPTN was shown to promote its binding to ubiquitin chains, while facilitating TBK1 activation, the retention of OPTN on mitochondria, and thereby efficient mitophagy, working as a positive feedback mechanism . The impIn 2018, Heo and colleagues further showed that upon mitochondrial depolarization, TBK1 phosphorylates the Ser72 residue of Rab7A in a Parkin-PINK1 dependent manner . This reThe phosphorylation of Rab7A was shown to be antagonized by PTEN, a dual-specific phosphatase that can dephosphorylate both lipid and protein substrates . It was A paper published in 2020 by Ritter and colleagues also reported the involvement of Rab7A phosphorylation in tumor progression . It had Collectively, it has been well established that the biological activity of Rab7A is regulated through phosphorylation by LRRK1 and TBK1, although it remains elusive whether there is a crosstalk between these two signals.In recent years, new aspects of the regulation of the Rab function by phosphorylation have become apparent, some of which suggest a role in diseases such as PD, ALS and cancer. This review focuses on recently published articles, with a particular focus on phosphorylation in the switch II region of Rab proteins. It is desirable for future studies to shed light on the more detailed role of Rab phosphorylation in regulating its function, and on the crosstalk between disease-related signal transduction and intracellular vesicle trafficking, in which Rab kinases may play an important role.


Sea-ice contamination in the antenna field of view constitutes a large error source in retrieving sea-surface salinity (SSS) with the spaceborne Soil Moisture Active Passive (SMAP) L-band radiometer. This is a major obstacle in the current NASA/Remote Sensing Systems (RSS) SMAP SSS retrieval algorithm in regards to obtaining accurate SSS measurements in the polar oceans. Our analysis finds a strong correlation between 8-day averaged SMAP L-band brightness temperature (TB) bias and TB measurements from the Advanced Microwave Scanning Radiometer (AMSR2) in the C-through Ka-band frequency range for sea-ice contaminated ocean scenes. We show how this correlation can be employed to develop: (1) a discriminant analysis that is able to reliably flag the SMAP observations for sea-ice contamination and (2) subsequently remove the sea-ice contamination from the SMAP observations, which results in significantly more accurate SMAP SSS retrievals near the sea-ice edge. We provide a case study that evaluates the performance of the proposed sea-ice flagging and correction algorithm. Our method is also able to detect drifting icebergs, which go often undetected in many available standard sea-ice products and thus result in spurious SMAP SSS retrievals. These instruments measure the electromagnetic passive microwave emission from the Earth’s surface and atmosphere at a frequency of 1.41 GHz or wavelength of 21 cm. Retrieval algorithms for these sensors, which turn brightness temperature (TB) measurements into SSS, have been developed and successively refined [ ESA’s SoThe retrieval accuracies of the L-band sensors strongly degrade in polar oceans. A major factor in this degradation is the decreased sensitivity of the L-band surface emission to SSS measurements at low sea-surface temperatures (SST). This results in a decreased signal-to-noise ratio in cold water (SST < 10 °C). To be specific, at colder SSTs, an error of +1 K in the TB measurement translates into an error of about –4 psu in the SSS retrieval. As such, improving SSS retrievals in or near polar regions has been identified as a major future science goal in ocean microwave remote sensing –6.ice, which is defined as the sea-ice concentration weighted by the antenna gain pattern [ice is the sea-ice concentration (SIC) at the Earth’s surface. The integration in B,c in ice of only 1% will result in a positive TB bias error of about +1.25 K, which, in turn, translates into a fresh bias error of about –5 psu in the SSS retrieval. At L-band frequencies, the sea-ice TB depends on ice-thickness, which can vary over location and season, in particular in the N hemisphere [ice in ice at the Earth boresight or within the main beam is zero. For both the Aquarius and the SMAP sensors, a sizeable portion of the antenna gain pattern G falls onto the sidelobes.In this context, SSS retrievals close to the sea-ice edge have turned out to be particularly challenging. Even very small sea-ice fractions in the antenna field of view result in large spurious salinity biases. The degree of sea-ice contamination in the antenna field of view can be characterized by the quantity g pattern :(1)gimisphere . It intrB,c in This discussion demonstrates the necessity for (1) developing a reliable procedure to detect small sea-ice concentrations in the field of view of the L-band sensor antenna; and (2) developing a correction for it. The second task amounts to estimating the contamination ΔTThe purpose of our study is to develop, train, and test sea-ice flagging and sea-ice correction algorithms for the SMAP SSS retrievals that can be implemented in future version updates. Our methods will not use external ancillary SIC fields due to their aforementioned insufficient accuracy in regards to low SIC near the sea-ice edge. Instead, both the SMAP sea-ice flagging and the sea-ice correction algorithms will use TB measurements from the JAXA Advanced Microwave Scanning Radiometer AMSR2), which takes measurements at frequencies between C-band and W-band and both vertical (V-pol) and horizontal (H-pol) polarizations. This idea was inspired by , which tThe spatial resolution of the SMAP footprint (≈40 km) and the lowest frequency AMSR2 frequency channel at C-band (≈50 km) are close. When multi-day aggregates are considered, in which several footprints are averaged together, then both sensors, SMAP and AMSR2, see approximately the same Earth scenes. Observations at the higher AMSR2 frequencies can be spatially resampled to the coarser C-band resolution.Compared to the SMAP L-band channels, the AMSR2 channels are insensitive or only very weakly sensitive to SSS. The AMSR2 ocean measurements are affected by sea-ice contamination but not by signals such as freshwater river plumes or melt run-off. This enables us to separate the spurious sea-ice signal from the real freshwater signals.ice in the SMAP 40-km footprint of about 1.5% or more. This is a much lower detection threshold than the sea-ice edge, which is commonly defined as the location where the SIC is 15%. The low detection threshold is mandated by the aforementioned high sensitivity of the SSS retrieval to sea-ice contamination. On the other hand, it is also essential to keep the false alarm rate as low as possible. False alarms will either result in unnecessary data loss or overcorrection. It is quite evident in The proposed sea-ice flagging algorithm is designed to detect sea-ice concentrations gThe vast majority of the scientific data users of the NASA/RSS SMAP SSS data utilize the eight-day or monthly time-averaged product from the Level 3 files. The Level 2 data are too noisy for most science applications. We have therefore decided to optimize our sea-ice flagging and sea-ice correction algorithms to perform best for the eight-day time-averaged SSS products. Thus, the analysis presented herein will focus on eight-day aggregate SMAP and AMSR2 TB observations.In our study, we will present results for the SMAP observations at the 40-km sensor resolution. The standard NASA/RSS SMAP SSS product has been smoothed to a spatial resolution of 70 km, which results in a noise reduction over open ocean scenes that makes the product more useful for most scientific applications . In rega2.2.1.2.1.1.B0(SMAP). It is calculated from the SMAP antenna temperature (TA) measurement after removing (1) antenna effects (antenna emissivity and cross-polarization contamination); (2) celestial radiation coming from the cosmic microwave background, galaxy, sun, and moon; (3) Faraday rotation in the Earth’s ionosphere; (4) atmospheric attenuation by oxygen, water vapor, and clouds; and (5) the wind induced rough surface emission [B0 maps.The source for the SMAP TB are the L2C files from the NASA/RSS SMAP SSS V4.0 Release , which hemission –15. We s2.1.2The source for the AMSR2 TB are the L1A AMSR2 TA measurements. Our study uses observations from the 6.93, 10.65, 18.7, 23.8, and 36.5 GHz V-pol and H-pol AMSR2 channels. We do not use the 2nd C-band channels (7.3 GHz) in our study as they contain no additional information to the 6.93 GHz channels in regard to developing our sea-ice flagging and correction algorithms. We also do not use the 89 GHz channels, as they are heavily affected by clouds and rain, which could introduce errors in our analysis. The observations of all channels are spatially resampled onto the largest footprint, which is one of the C-band (6.93 GHz) channels (62 km 35 km) . The spaB,TOA(AMSR2), using the calibration procedure outlined in [B0(AMSR2). In the calculation of the atmospheric attenuation, we use the simplified bulk expressions derived in [0(AMSR2) of the AMSR2 channels are determined by:S is the SST in Kelvin.The AMSR2 TA are first transformed into the top of the atmosphere (TOA) TB, labeled Tlined in . We empllined in  and the lined in  for the rived in  that depThe AMSR2 observations are screened for degraded conditions by removing measurements that are contaminated by land, sun-glint, known sources of radio frequency interference (RFI), and rain. When removing land contaminated measurements, we require that the land fraction within the AMSR2 6.93 GHz footprint is less than 1%.B,TOA(AMSR2), and the specular surface E0(AMSR2).Finally, the resampled AMSR2 TB are averaged in space and time into 8-day aggregate 0.25° maps, which comprise the same time periods as the SMAP TB map. When gridding the AMSR2 swath observations into regular 0.25° maps, we use a simple straight average (drop in the bucket). We will create and use separate sets of maps for the TOA TWe note that the value of 1% for the land fraction threshold is the maximal land fraction that is allowed within a single AMSR2 C-band footprint. In each 0.25° cell several of these footprints are averaged together, so the actual land contamination is in general considerably less than 1% in the 0.25° cells, even at locations closest to the coast or near islands. The land fraction does not change over time, so by careful inspection of the cases used in this study we can check that land contamination is not a major error source in our analysis. We refer to the 2.1.3.B0,exp, is the result of the RTM calculation of the expected specular surface emissivity E0,exp multiplied by TS. The expected surface emissivity E0,exp is obtained from the Meissner–Wentz model of the dielectric constant (permittivity) of sea-water [B0,exp for SMAP, and expected E0,exp for AMSR2.The expected specular surface TB, labeled Tea-water ,20,22. OB0,exp are the basic metric for training and testing our proposed SMAP sea-ice flagging and sea-ice correction algorithms. At L-band frequencies, both TB0,exp and E0,exp depend on SST and SSS and, thus, their calculation requires an ancillary reference SSS field   as ancilCross Calibrated Multi-Platform (CCMP) V2.0 wind speeds and directions –28 for tAtmospheric profiles for pressure, temperature, relative humidity, and cloud water density as well as total columnar water vapor and total columnar cloud water from the General Forecast System (GFS) of the National Centers for Environmental Prediction (NCEP) . These fPrecipitation rates from the Integrated Multi-satellitE Retrievals for the Global Precipitation Mission (IMERG) Version 6  for rainWe note that these ancillary fields are the same that are already being used in the V4.0 NASA/RSS SMAP salinity retrieval algorithm for the purpose of turning measured SMAP TOA TB into surface emissivity. Extensive analysis has been conducted and shown that they are well suited for that purpose ,31.2.1.5.Our study is based on several training and test sets of SMAP and AMSR2 data, which are summarized in As discussed in The training of the algorithms turns out to be very robust and using scenes from the Northern hemisphere is not harmful. For training the sea-ice detection and flagging algorithm, we have found that it is actually slightly beneficial to use Arctic scenes, but only if the observations in known areas of freshwater river outflows are discarded.2.1.6.We impose a climatological monthly sea-ice mask as a-priori condition for the sea-ice detection and correction algorithms. This climatological sea-ice mask was compiled from many years of historical SIC data before the launch of SMAP and AMSR2. Its sole purpose is to provide a simple indicator for locations in the ocean where sea-ice can form. The mask also includes areas where drifting icebergs have been detected. 2.2.2.2.1.The first and biggest step of the sea-ice detection and flagging algorithm employs the Fisher Discriminant Analysis, which is a well-established method in pattern classification for distinguishing two different classes of data points in a higher-dimensional space. 1 = 0.4 K, e2 = 2.0 K, e3 = 4.5 K. The RMS difference for 8-day aggregate ΔTB0(SMAP)V-pol over open ocean scenes is found to be about 0.2 K, so any observation that falls within Class 2 is very likely contaminated by sea-ice. The sea-ice detection threshold e2 of 2.0 K corresponds to an approximate sea-ice concentration gice of 1.5%. If the discriminant analysis determines that an observation falls into this class, then it is flagged as sea-ice. Observations that fall within Class 1 might still contain some sea-ice contamination, and we address them in the second step of the sea-ice flagging algorithm. It is necessary to leave a large enough separation between the upper threshold e1 of Class 1 and the lower threshold e2 of Class 2. If the values for these thresholds are too close together, the discriminant analysis is less effectively able to distinguish between the two classes, which would result in an increased number of missed detections, false alarms, or both. The reason for imposing an upper threshold e3 in the definition of Class 2 will be discussed shortly.When training the discriminant analysis, we draw data from two classes both of which are subsets of the 4 training sets listed in X of the 10 AMSR2 TB channels, each of which represents a point in a 10-dimensional data space. We consider two cases for the AMSR2 data points X: Case 1 uses the TOA AMSR2 TB. Case 2 uses the measured–expected AMSR2 surface emissivities:eff = 273.15 K serves to convert the dimensionless emissivity values into quantities with dimension K. The index k in The inputs to the discriminant analysis are the measurements W within this 10-dimensional space, which results in the best separation between the two classes after the data are projected on it. The elements of the projection vector W can be regarded as optimal weighting coefficients of the 10 AMSR2 channels. W that were determined from the training Sets 1–4 from B0. The discriminant is given by the projection of the data vector X onto the projection direction W, which is the scalar product WT · X of the two vectors. For the determination of the criterion to distinguish between the two classes, we plot the probability density functions (PDFs) or histograms of the discriminant WT · X for each class  exceeds the value e3 into Class 2, thus classifying it correctly as a sea-ice contaminated observation.Including an upper threshold value e2.2.2.2 in the class definition (5) would achieve that, but it would also result in an increased number of false alarms, which is not desirable. In order to catch the majority of these low SIC cases, we add a NN flagging as a 2nd step of the sea-ice detection algorithm. The basic assumption is that most of the sea-ice contaminated observations that went undetected in the 1st step are in close spatial proximity to at least one of the events that have already been flagged. Our sea-ice detection flags both the nearest neighbors and the next to nearest neighbors of any cell that the discriminant analysis has put into Class 2. This amounts to flagging the 24 surrounding cells of the 0.25° grid together with any cell from Class 2.The discriminant analysis used in the 1st step of the sea-ice detection algorithm is designed to safely detect sea-ice contaminations of 1.5% or more as part of Class 2. However, observations that the discriminant analysis does not put into Class 2 can still be contaminated by sea-ice and thus might need to be flagged as well. Lowering the threshold value e2.3.ice (OSI-SAF) exceeds about 1%.The combined discriminant and NN sea-ice flagging scheme allow us to define sea-ice zones, which classify the severity of sea-ice contamination for each observation. This will become helpful for developing and evaluating the sea-ice correction algorithm. 2.4.B,c for the sea-ice flagged observations , which act as noise sources in the detection and correction algorithms. B,0(SMAP V) and the scaled AMSR2 6.93 GHz V-pol specular TB bias λ ΔTB,0(AMSR2 6V) for the test set scenes  and the sea-ice correction terms ΔTB,corr,1 and ΔTB,corr,2 that were derived in A pivotal point for the performance of the proposed flagging and correction method is the very good correlation between the SMAP and the AMSR2 measured–expected TB for the 8-day aggregate observations near the sea-ice edge. The ΔTt scenes  within st scenes . Similart scenes  for the We note that the lowest frequency AMSR2 channels (C-band and X-bands) correlate best with SMAP, though the higher AMSR2 channels (Ku–Ka-bands) still contain valuable information when it comes to sea-ice contamination, and it is, therefore, beneficial to include them in our method.3.2.ice in each zone.B,0 in each of the zones. The values show that this goal has been largely achieved. It is also encouraging to see reduced values for the standard deviations. The relative improvement compared to the uncorrected cases is strong for Zone 3 observations and even stronger for Zone 4 observations. We should note, however, that the remaining errors after the correction for Zone 3 and Zone 4 will still result in large errors in the SSS retrievals. A TB error of 1 K translates into an SSS error of about 4 psu in cold water  that remain after the sea-correction is applied in more detail in 3.4.3.5.Very large drifting icebergs are a potential source of serious errors in the SMAP SSS retrievals. Even though icebergs are not sea-ice, the electromagnetic properties of sea-ice and icebergs result in a similar degree of contamination in the SSS measurement. The a-priori climatological ice mask  delibera2 of undetected contaminated SMAP SSS data over the course of several weeks. A good example of this detection and correction is provided by the very large iceberg A-68 , which wWe note that neither of the products ,10 shows4.This section discusses the uncertainty and error sources in our methodology and its limitations.Radiometer noise in the AMSR2 TB measurements is a source of error. However, the noise in the individual measurements is strongly reduced in the eight-day averages. Thus, the impact of AMSR2 radiometer noise on our detection and correction algorithms is small.ice and decreasing distance from the sea-ice edge. The ancillary data sets for these parameters are Level 4 products that rely on microwave satellite observations as input. When approaching the ice edge, the microwave satellite observations are flagged, and, thus, fewer satellite input data become available. This can result in lower accuracy of the ancillary Level 4 product [A larger uncertainty source are errors in the external ancillary fields, which might not get suppressed in the eight-day averaging. This impacts the algorithms from Case 2, which use the ancillary fields to remove atmospheric and wind roughness effects from the AMSR2 TOA TB. We expect the errors in the ancillary fields for SST, wind speed, and direction to increase with increasing sea-ice fraction g product .One of the major assumptions when using the AMSR2 TB to flag and correct the sea-ice contaminated SMAP TB was that the spatial resolutions of the SMAP and the AMSR2 TB measurements are similar. When averaged over eight days, the different orientations of the antenna footprints at a certain location have less of an impact. This means that SMAP and AMSR2 see approximately the same Earth scene. That assumption is of course only approximately fulfilled, and in reality, there is some spatial mismatch between the SMAP and the AMSR2 observations. It is possible to reduce this uncertainty source by resampling the SMAP and all the AMSR2 channels to the same target location and the same spatial resolution by employing the Backus-Gilbert optimum interpolation technique . This reThere are scenarios that can result in a degradation of the good correlation between the SMAP and the AMSR2 TB. The most important one is thin sea-ice, which impacts the higher AMSR2 frequencies more than the lower ones or the SMAP L-band. We note that, at L-band, the sea-ice penetration depth is approximately 0.5 m, and the penetration depth grows with the electromagnetic wavelength. Large areas of thin ice tend to form in the Arctic oceans during fall. In the S hemisphere, this problem is less prevalent. Further studies are necessary to examine if and how much this poses a problem for our flagging and correction method.Undetected RFI in the AMSR2 TB also constitutes an error source.A problematic case is sea-ice close to the coast, which can result in simultaneous land and sea-ice contamination of the SMAP antenna field of view. Our proposed sea-ice detection and correction algorithms were not designed to handle this case. Likewise, we expect that the sidelobe correction for land contamination that is currently applied in the NASA/RSS SMAP SSS retrievals ,14 is noThe flagging method is not able to detect observations of very small sea-ice fractions that are significantly lower than the detection threshold of the discriminant analysis (1.5%) and that are not NN or next to NN of an observation that has been flagged by the discriminant analysis.B,0(SMAP) in each sea-ice zone, which they do  , which isupport data


Recent studies suggest that higher Body Mass Index (BMI) is associated with reduced inhibitory control in contexts of palatable food. However, due to limitations of previous studies, it remained the question whether this reduction is specific to food contexts, and whether it generalizes to other contexts of reward, such as money. This main question was addressed in the current study. In addition, we explored the effect of maladaptive eating and stress regarding inhibitory control across the contexts that differed in terms of reward.In total, 46 participants between 19 and 50 years old  with an average BMI of 23.5 (SD = 3.9) participated. Participants filled out questionnaires and performed a go/no-go task (indexing inhibitory control) with three conditions .Relatively high (above median) BMI was associated with challenged inhibitory control in the food relative to the neutral context, but not in the money relative to neutral context. Explorative analyses suggested that maladaptive eating and stress were associated with reduced inhibitory control in the food context. Only rumination was associated with reduced inhibitory control in the money context.The effects of BMI, maladaptive eating behavior, and stress on inhibitory control were specific to the food context, and did not generalize to a non-intrinsic reward condition, operationalized with money pictures. Our results imply that (research on) interventions directed at improving inhibitory control in relation to overweight and obesity, should consider food-reward context. Overwei30 kg/m2 . Certain30 kg/m2 , 5, and 30 kg/m2 . In that30 kg/m2  and unco30 kg/m2 , as well30 kg/m2  and rumi30 kg/m2 , especia30 kg/m2 . The opeThough several studies have focused on the effect of inhibitory control on BMI, other studies suggest that increased BMI may predict challenged inhibitory control, suggesting a bidirectional causal relationship. Pertaining to the mechanism of the latter, normally, perceiving reward related stimuli  results in increased striatal dopaminergic neurotransmission, and makes approach behavior more likely , 13. MeaStudies that focused on behavioral indices of inhibitory control in relation to BMI are consistent with the above. Firstly, no consistent relationship between BMI and inhibitory control in neutral contexts is evidenced, but the relationship depends on both subject and environmental (reward context) variables . With reOne open question that has not yet been thoroughly addressed, is whether the challenged inhibitory control is specific to a food context, or whether it generalizes to other reward contexts as well. The latter may be expected following incentive sensitization theory which predicts that over time, sensitization to one type of reward(-related stimulus) extends to other reward(-related) stimuli as well through cross-sensitization , 19. TheWith respect to eating behaviors, it should be emphasized that especially uncontrolled/binge eating  and emotIt should be noted that stress may also have a moderating role regarding the relationship between BMI and inhibitory control, which may be most prevalent in a food context. In other words, the relationship between BMI and inhibitory control may vary as a function of stress. The study of Nederkoorn et al.  is notewIn the current study, we measured inhibitory control using a go/no-go task modeled after Wessel , that inIntegrating the above, it was hypothesized that higher BMI as well as related maladaptive eating patterns would be associated with reduced inhibitory control in reward contexts relative to a neutral context. In addition, the same was hypothesized with respect to rumination and stress, while stress was also hypothesized to moderate the relationship between BMI and inhibitory control, especially in a food context.2), . In total 34 participants had normal BMI  with a gender distribution of 40% males and 60% females respectively, and 12 participants  presented with overweight (n = 8) or obesity (n = 4). The study was approved by the Research Ethics Committee and conducted following the declaration of Helsinki. Written informed consent was obtained from all participants.Participants were recruited predominantly via advertisements on social media . Participants were excluded if they had any known current mental disorder, if they currently used drugs affecting cognitive functioning, if they had color blindness and if they were pregnant. Participants had to be between 18 and 50 years old. Participants who did not complete the experiment were excluded. The final sample for final analyses consisted of in total forty-six participants , ranging in age from 19 to 50 years old  and Body mass index BMI (kg/mThe go/no-go task  was emplThe participants eating behavior was assessed by the short version of the three-factor eating questionnaire (TFEQ-R18). Previous research has shown that the TFEQ-R18 yields a good measure of uncontrolled and emotional eating and has a good factor structure and reliability with Cronbach’s alpha ranging between 0.78 and 0.94 . The TFE(“What am I doing to deserve this?”) when experiencing low mood. Items are answered on a 4-point Likert-type scale, ranging from 1 "almost never" to 4 "almost always"; higher scores reflect higher rumination. In our study Cronbach’s alpha was 0.943.The RRS is one of the most popular instruments to assess ruminative responses , and hasThe DASS-21 is a widely used instrument to assess levels of depression, anxiety and stress by self-report, also in relation to clinical symptomatology. The instrument has a good overall reliability with a Cronbach’s alpha of 0.91, 0.84 and 0.80 for the Depression, Stress and Anxiety subscale, respectively . Each ofThe assessments were completed online, via Psytoolkit , 35. AllRepeated measures ANCOVAs were performed for the primary analyses, with alpha level set at 0.05. Analyses were similar to the ones performed in Houben et al. , and oneDescriptive data for age, BMI, eating behavior, rumination, stress and subjective hunger level is shown in Table p = 0.622, partial η2 = 0.011. The secondary analysis, including BMI as categorical variable based on split by median (= 22.43) yielded a BMI (above median/below median) × condition  interaction that trended to significance, F = 2.73, p = 0.071, partial η2 = 0.058. Post-hoc analyses indicate that above median BMI relative to below median BMI was associated with reduced inhibitory control in the food condition relative to the neutral condition, F = 4.12, p = 0.049, partial η2 = 0.086  = 1.01, p = 0.369, partial η2 = 0.022.The relationship between BMI and inhibitory control did not significantly vary as a function of condition , F2,88) = 0.48, ,88 = 0.4As shown in Table Our results indicate that higher BMI is associated with reduced inhibitory control in a food context relative to a neutral one. In addition, explorative correlational analyses seem to indicate that maladaptive eating and negative effect were associated with reduced inhibitory control in the food context. In contrast to our hypotheses, effects with the exception of rumination, did not generalize to the money context.In a previous study, it was already reported that higher BMI was associated with reduced inhibitory control in a food context relative to a neutral context. However, one important limitation challenged the interpretation of effects. More specifically, in Houben et al.  counterbIn line with previous reports , the expAs mentioned, it was expected that the negative association between BMI, maladaptive eating, stress and rumination on the one hand and inhibitory control on the other in the food context relative to the neutral context, would be mirrored in the contrast between the money context relative to neutral context. However, except regarding rumination, our results did not support that notion. One might argue that this implies that the challenged inhibitory control is restricted to food contexts and may not generalize to other reward contexts. However, one limitation of the current study may be the operationalization of the non-food, reward condition operationalized by stimuli representing money. We employed the money condition because stimuli of money are obviously different than food, but are also known to trigger primary reward related activity . HoweverOn the other hand, results from Godefroy et al. , may sugIt should be noted that the relation between BMI and inhibitory control in the food context relative to the neutral context was only statistically significant in our exploratory analysis when including BMI as a categorical (median split) variable. Following Houben et al. , we deciOne other limitation, at least perceptually, is that the study was conducted online. However, it should be emphasized that recent studies consistently show that online cognitive psychological experiments can generate reliable and valid data, comparable to those conducted in a controlled lab environment , 42. Of Furthermore, the use of the go/no-go task as opposed to the stop signal task, to measure inhibitory control, could be viewed as a limitation. Firstly, there is substantial heterogeneity of go/no-go task implementations in terms of task specifics. Until recently, it was not entirely clear what task characteristics were essential to induce inhibitory control. However, as shown in Wessel et al. , a fast-Our results indicate that higher BMI, maladaptive eating patterns  as well as rumination are associated with challenged inhibitory control when perceiving palatable food related stimuli relative to neutral stimuli. We could not confirm that the effect generalizes to a different, and non-intrinsic reward context operationalized by money. The results have clinical implications in that interventions aimed at improving inhibitory control may benefit from a focus on inhibitory control in a context of palatable food. Specifically, it has been shown that strengthening inhibitory control can assist individuals with obesity in regaining control over the consumption of palatable foods, as well as making them less susceptible to the temptations of palatable food , 44. Our