Clinical outcome after successful reorientation of an upside-down implanted DMEK  graft 4 weeks after initial transplantation.A 71-year-old woman presented with Fuchs’ endothelial corneal dystrophy for DMEK. After initial DMEK the donor graft was fully attached and well centred during intracameral gas filling. When the gas bubble was fully resorbed the graft started to detach. Therefore, two intracameral gas injections were consecutively performed. During the second re-bubbling, an upside-down orientation was observed and so the graft was flipped, centred, re-attached and finally stabilized by an intracameral gas bubble. Three weeks after reorientation slit lamp examinations showed a well centred and attached graft, endothelial cells that started functioning and a patient’s visual acuity of 20/40. Visual acuity increased to a 20/32 vision in the observed eye three months later and further improved to 20/20 6 months after reorientation and stayed stable between 20/32 and 20/20 during the remaining 15 months of follow-up, with a clear and well-attached graft.Reorientation of an upside down DMEK graft was successful even 4 weeks after initial DMEK. Visual recovery and endothelial cell count increase were stepwise noticed during the first 6 months and 15 months after reorientation, respectively. Finally a favourable outcome with 20/32 to 20/20 vision at least 6 months after graft reorientation was achieved. Therefore, restoring full graft function could last several weeks or even months following (late) reorientation of an upside-down DMEK graft. In Fuchs’ endothelial corneal dystrophy (FECD) or other corneal endothelial disorders reduction of postmitotic endothelial cells results in a loss of cellular hexagonal morphology  and alteration in cell size. As a result of those alterations, the remaining endothelial cells are not able to maintain the normal pumping and leaking function. Corneal thickness and glare sensitivity increase, while visual clarity decreases.Over the past years, Descemet Membrane Endothelial Keratoplasty (DMEK) has evolved rapidly as a preferable alternative to penetrating keratoplasty in patients with endothelial cell disorders like FECD. DMEK enables selective transplantation of the endothelial monolayer including the Descemet membrane, promising favourable functional and anatomical results .Graft detachment is a potential complication after DMEK. Therefore, surgical interventions like air/gas re-injection inside the anterior chamber (graft re-bubbling) or graft reorientation of an upside-down graft can be considered. In cases of a detached graft and an assumed proper orientation of the transplant re-bubbling is the first treatment option to manage partially detached grafts after DMEK. The most important requirement for a successful reattachment is a correctly orientated graft .A large retrospective multicentre real-life study investigated 2485 eyes after DMEK and found a re-bubbling rate of 20.1% .Dirisamer et al. investigated the type of graft detachment in 150 consecutive DMEK cases and described clinical signs of an upside-down orientation of the graft like persistent stromal oedema and spike-shaped fibrous lesions between the donor and recipient interface .This is the first report describing a successful graft reorientation of an upside-down DMEK graft even 4 weeks after initial DMEK.A 71-year old woman presented to our eye clinic with bilateral FECD. The patient’s medical history revealed previous cataract surgery. No other prior ocular surgery was performed in the past and no other ocular diseases were known. The woman had polymyalgia rheumatica and psoriasis arthritis treated with oral prednisolone 5 mg  once a day combined with oral pantoprazole 40 mg  and subcutaneous adalimumab 40 mg  biweekly. Due to a previous stroke without any residual deficits, the patient was treated with aspirin 100 mg daily .Best-corrected distance visual acuity (BCDVA) was assessed in the decimal scale and converted to Snellen equivalent for reasons of better comprehensibility and clarity. The preoperative BCDVA was 20/63 in her right eye and 20/63 in her left eye.DMEK was performed on the right eye using a liquid bubble dissection technique  for lentThe patient was instructed to maintain a supine position as long as the gas bubble was present. Examinations revealed a well-positioned transplant with a clear centre and only few Descemet’s membrane folds without any signs of detachment under the gas bubble on the first day after surgery. During the following days, most of the gas tamponade was absorbed, and progressive stromal oedema observed. Descemet’s membrane folds and subtotal Descemet detachment have occurred. A second air-gas mixture bubble was injected inside the anterior chamber on the sixth day after the initial surgery. After re-bubbling the patient showed a stable, well-centred graft with an air-gas bubble inside the anterior chamber and was discharged home. Three weeks after initial surgery the patient presented with blurred vision (BCDVA counting fingers), stromal corneal oedema, Descemet’s folds and Descemet’s membrane detachment infero-temporally. We discussed our findings with the patient and the shared decision was made to perform a second re-bubbling with controlling the orientation of the graft. Twenty-nine days after initial DMEK the patient presented for the planned intervention. Prior to the second re-bubbling, the patient achieved 20/600 vision in the right eye and presented with unchanged corneal findings. Before the injection of another air-gas mixture bubble, the surgeon noticed the unusual behaviour of the graft. The graft was detached from the cornea and checked again for an upside-down orientation. This was done by looking at the double roll of the graft which seemed to face downwards and so the surgeon concluded that the graft could be in an upside-down position. Therefore, the surgeon reorientated the graft by inversion of the entire transplant and injected the air-gas mixtureinside the anterior chamber after unfolding the graft a2a).Marked reduction in corneal clearance, increased corneal thickness, and a reduced visual acuity should be considered as a sign of the upside-down orientation of the graft.Fernandez Lopez described a higher rate of graft detachment, increased graft stiffness and/or fibrosis when re-bubbling was performed several weeks after initial DMEK . In our An upside-down orientation of the DMEK should also be considered if re-bubbling is not sufficient and an inverse corneal clearance pattern is observed. Our case demonstrated, that even 4 weeks after initial DMEK a successful graft reorientation is a possible treatment option with good visual outcome .


In order to extend the shelf life of the fruit, improve appearance, and to keep all nutrition properties of the plum from diminishing, edible coatings comprised of wheat starch and wheat starch–whey protein isolate (in ratio 80/20) were created. Stand-alone films were produced to assess properties which helped to understand the phenomena occurring on the surface level of coated plums. The properties of coatings based on starch are similar to starch coatings containing oil because the natural epicuticular wax layer of plums merges with coating materials. Adding oil doubled the contact angle value and the dispersive component of the surface tension. The workings of adhesion and cohesion, spreading coefficient, water absorption, water content, and solubility in water of the films decreased. Similar processes were observed on the fruits’ surface. In appearance, the coating process is similar to polishing the plum surface for removing crystalline wax. The color parameters of coated fruits did not significantly change. Newly formed bonds or interactions established between starch, whey proteins, water, glycerol, and oil are displayed by Fourier transform infrared (FTIR) analysis. This work revealed how the interactions between the epicuticular wax on the fruit’s surface and the hydrocolloid-based coatings affect the efficiency of the coatings. Biodegradable packages have a significant impact on the environment by helping to reduce waste. Total biodegradability is the highest advantage of these materials. Films and coatings can replace conventional packaging in many applications, such as disposable packaging materials, single use bags, cups, plates, containers, egg cartons, lamination coatings, etc. ,2,3. PreHydrocolloids and lipids are two types of biomolecules generally used for the preparation of biodegradable packages. Among all of them, starch is one of the most promising materials from which to develop biodegradable films. Starch has excellent film-forming and gas barrier properties in dry conditions . It alsoOnce applied onto the product surface, the coating materials are one of the food components. They can occur as single or multilayers. Coatings used to reduce transpiration losses in citrus fruits, pears, plums, and apples consist of one kind of film-forming material because the second material (wax) naturally occurs on the surface of the fruit ,12,13.Compared to protein-based coatings, the barrier property of starch-based materials, especially against water vapor, is less pronounced . Starch Plums have a natural wax on their skins . This thFor this work, plums were chosen for their high nutrition value, and also their relatively short shelf life. The skin can be consumed with the flesh, so plums are an excellent material for covering with edible coatings. The main objective of these investigations was to demonstrate the interaction between starch-based coatings and components of the fruit surface. Physicochemical analyses were carried out to study their effects on selected properties of the coating material. Stand-alone films were made as a model for understanding the processes which cannot be measured on the surface of the plum directly.The surface and the cross-section pictures achieved from environmental scanning electron microscopy (ESEM), as well as light and polarizing microscopies are presented in Photographs of coated plums showed good adhesion of the starch–oil–starch films e–j probaPrunus domestica) in their experiments: cv. Hanita and cv. Ortenauer. One of the goals of the work was to compare color parameters of untreated and polished plums, where the epicuticular wax layer was removed manually. Results for color parameters obtained by these authors show that plum species played a significant role . Nevertheless, L, a, and b values for untreated fruit were lower than that for cv. Jojo plums, i.e., 37.3, 2.35, −13.9, respectively. The values after taking off the epicuticular wax were even less for lightness: 28.1 and higher for redness; 2.86 for blueness; −5.0 for untreated plums of the same variety. Total color difference (ΔE) was under the value 2.3, so the slight difference in color was not visible for the human eye [Color parameters  given in uman eye . This fauman eye , which mScanning microscope observations revealed round drops on the surface. The same round drops of oil are visible on the starch films with rapeseed oil, and on plum surfaces a few hours after being coated in starch and starch–whey protein film-forming solutions d,i. So, Colocation between starch-glycerol systems and waxes explains why the formation of a coating of two opposite materials (polar and nonpolar) is possible. Moreover, surface tension is the lowest for starch films with an oil addition, which thereby means that fat causes the higher stability than is the case of wheat starch and whey protein systems. It is also confirmed by absorption. The value of starch matrices is about one-third higher than for starch–oil matrices and films with proteins. This means films with oil addition have lower water affinity than pure starch films and starch films with 20% of proteins addition ,29,30. SLooking on sorption isotherms , it is nFourier transform infrared (FTIR) spectroscopy is a technique which illustrates the shifting of bonds by collecting high spectral resolution data over a wide spectral range. Hence, the coatings of two opposite materials, starch–glycerol systems (or starch–whey protein–glycerol films) and waxes (polar and nonpolar), can be explained by the results obtained from infrared spectroscopy.w/v). At 3430 m−1 (not showed onto the graph) is located a broadband with a very strong character. OH-groups come from the absorbed starch polymer, from water, and from glycerol corresponding with this peak. Thus, broadband with wavenumbers 2928 m−1, 1655 m−1, and 1373 m−1 either have weak character and they were assigned with hydrogen bond, the δCH, and the C=O, respectively. Peaks with wavenumbers of 1165 m−1 and 1084 m−1 are attributed to CH and δCO groups but, in contrast to previous bonds, they have stronger energy. Additionally, a peak located at 980 m−1 has energy and is associated with δC–O stretching vibrations [−1 [−1 in the spectrum which is assigned to free N–H groups [−1 in the spectrum of whey protein isolate is corresponding to hydrogen bonded N–H stretching [−1 are characterized for amide I. So, the peak at 1643 m−1 also correspond to the C=O stretching vibrations of amide I [−1. Aggregated proteins show a peak assigned to 1600 m−1 which is characteristic of intermolecular β-sheets [−1 is attributed to the C−N stretching and N−H bending of vibrations from amide II [−1 attributed to N−H bending and C−H stretching vibrations, it is amide III region [−1 are assigned to β-sheet structures [The most important interaction between molecules which describes the wheat starch system is hydrogen bond . The FTIions [−1 . In turnH groups . The pearetching . Wavenum amide I . The N−Hβ-sheets . Peak loamide II ,43. Thusamide II . WavenumI region . Thus, tructures .−1 which are associated with the vibration of the skeleton C–C. The broadband at 1045 102 m−1 corresponds to the stretching of the C–O linkage in C1 and C3, and the peak at 1117 m−1 is attributed to the stretching of C–O in C2 [Spectrum FTIR of pure glycerol reveals bands at wavenumbers 850, 925, and 995 m–O in C2 .−1 can be attributed to C–O stretching, are shifted to 1020 m−1 . Moreover, the peak located at 2908 102 m−1, corresponding to C–H vibrations, also shifted to a higher position to 2932 cm−1 for starch and whey protein films, and to 2938 m−1 for the films with oil. Broadband assigned to O–H bond at wavenumber 3320 m−1 is shifted to 3338 m−1 for starch and starch–whey protein films, and to 3356 m−1 for oil starch systems. The addition of glycerol influences the hydrogen bonding interactions among starch and glycerol, starch–whey protein and glycerol, and starch–oil and glycerol systems. Accordingly, the decreased intensity of the peak at 3293 m−1 is attributed to reactions between groups of starch and NH groups of whey proteins which shift the number of NH groups [The effect of glycerol, water, and oil can be analyzed by comparing the spectra of wheat starch powder and glycerol to the spectra of starch films of 50% glycerol and spectra of wheat starch powder, whey protein powder, and glycerol with starch–whey protein films with 50% of glycerol. Comparing wheat starch and whey protein isolate powders to the films made from 5% starch and 5% starch–whey protein (in ratio 80/20), in both matrices with 2.5% of glycerol, as seen on H groups .Wheat starch was supplied by Hortimex . The whey protein isolate  BiPRO was obtained from Davisco Foods International Inc. . Anhydrous glycerol (99.9% of purity) was purchased from Sigma-Aldrich .Prunus domestica L., cv. Jojo) were harvested from the technology garden (orchard) of Leibniz-Institute for Agricultural Engineering  at the commercial ripening stage and transported immediately to the laboratory. Afterwards, 180 plums (from 4 trees), with an average weight of approximately 30 grams, were split into three groups for the following treatments: The first group was the control, the second group was coated with a 5% film-forming solution of 80% wheat starch and 20% whey protein isolate, and the third group was coated with a solution of 5% of wheat starch. All fruit were weighed on a sensitive scale CPA225D  and stored in partly covered plastic trays in a climatic chamber  under controlled environmental conditions of 5 °C, 80% relative humidity (RH), and natural convection for 1 day before undergoing the coating process.Plum fruits (w/w). Glycerol was used as a plasticizer at 50% w/w of biopolymer dry weight . Wheat starch film-forming solutions were prepared by dissolving 5 grams of whey starch powder in 100 mL distilled water. Whey protein film-forming solutions were also prepared by 100 mL dissolving 5 grams of whey protein isolate in distilled water. The solutions were heated separately in a water bath under a 12 s−1, stirring at 85 °C for 30 min to denature the whey protein and to obtain complete gelatinization of the starch. Then the film forming solutions were cooled down to 40 °C. The plasticizer (2.5 g in each solution) was added and, thereafter, the solutions were mixed in the aforementioned ratios.Film-forming aqueous solutions were mixed by an Ergo Mixx blender  and wheat starch and whey protein isolate were added in the following proportions: 100/0% and 80/20% , m is the mass of the film-forming solution (g), ρ is the density of the film-forming solution (g cm−3), e is the mean thickness of the coating layer on each fruit (cm), FM is the mass of the fruit (g), A is the surface area (cm2) of the fruit.The mean thickness of the coating was calculated from the difference in mass between coated and uncoated fruit, the density of the coating material, and the surface area of the fruit:Equation (2) was established from preliminary experiments. Thirty-six plums were scanned using a camera supported by a 3D ScanBook scan system along with ScanWare Enterprise 3.8 software . The measured surface area was set into relation to fruit mass. This relation found by a regression analysis is valid only in the range between 30–80 g of fruit mass.The relevant mass of film-forming solution covering the fruit surface was determined from the difference in weight of the plums before and after applying the coating (dipping).Film thickness was measured with a PosiTector 6000  digital micrometer to the nearest 1 µm in 0–100 µm range and to the nearest 5 µm in the 100–1000 µm range. Prior to film thickness measurements, the electronic gauge was calibrated at 74 and 139 µm using standards to be close to the thicknesses of the samples. The thickness of each film was measured at five points at the center of each film and at four points around the perimeter. An average value was used in the calculations. At least ten replications of each formulation were made.Contact angle (θ) is the angle described as a relationship between the surface tension and one of three phases: Liquid phase L, solid phase S, or vapor phase V according to the Young equation . The conThe surface tension of the liquid tested (γL) was measured by the sessile drop method and Laplace–Young approximation ,51. The The surface tension, or surface free energy (t method  as evideIn Equation (4), appear two unknowns Work of adhesion , work of cohesion , and spreading coefficient  were calculated using  equationThe water content was measured in samples of 2 × 2 cm, by determination of the weight loss of the film after drying at 105 °C for 24 h in a drying oven UT20 , and is expressed as gram of water per gram of dry matter. All experiments were performed in triplicate.The absorption index was measured to assess the impact of immersion on water absorption. The samples were prepared from five different films of the same kind. They were cut into 2 × 2 cm pieces and weighed on an analytical balance CPA224S-OCE  with an accuracy of 0.0001. They were then immersed in distilled water (25 °C) for 2 min. Wet samples were wiped with filter paper to remove excess liquid before weighing. The amount of adsorbed water was calculated in percentages. The measurement was repeated for each type of film three times, and the average was taken as the final result.Water solubility was determined according the  method. The sorption isotherm of films was determined at 25 °C. Samples of films were cut into small pieces (2 × 2 cm) and weighed to the nearest 0.0001 g in preweighed vials. Films were stored up to equilibrium in ten desiccators, each containing a saturated salt solution which fixed the relative humidity. A wide range of RH was selected: Lithium chloride (11%), potassium acetate (22%), magnesium chloride (33%), potassium carbonate (43%), magnesium nitrate (53%), sodium nitrite (65%), sodium chloride (75%), ammonium sulfate (81%), and ammonium dihydrogen phosphate (93%).Film samples were weighted periodically, up to the equilibrium was reached and checked up to 9 months. The final (equilibrium) water content was measured by drying the films at 105 °C for 24 h. The amount of water absorbed is expressed as grams of water per gram of dry matter. Measurements were done in triplicate for each film recipe.−1 s−1 Pa−1) was calculated using the following equation:m/Δt is the moisture loss per unit of time (g s−1); A is the film area exposed to moisture transfer (8.04 × 10−4 m2); e is the film thickness (m); and, Δp is the water vapor pressure differential between the two sides of the film (Pa). Measurements were performed at least three times for each differential tested.The water vapor permeability of the films was measured gravimetrically according to  who adap−1 at atmospheric pressure. The increase in pressure in the downside chamber during the test period was assessed and displayed by an external computer. Data was recorded and permeance was calculated by GDP-C software (with temperature compensation). The sample temperature (25 °C) was adjusted using an external thermostat Haake F3 with water bath K . The desired RH was regulated in an external saturation system (53% and 75%), so humidified oxygen gas circulated in the permeation cell.The oxygen permeability was measured using the manometric method according the  standardE) and color chroma according to the following equations [L*, a*, b*, and C are the color parameters of a white standard used as the film background .The color of the films was determined using a colorimeter CR-300  using the CIE LAB color parameters: L, from black (0) to white (100); a, from green (−) to red (+); and b, from blue (−) to yellow (+) . The colquations :(10)ΔE=, using a colorimeter CM-2600d . Three determinations were performed.Film microstructure was observed using an environmental scanning electron microscope ESEM XL-30 . A 0.5 × 1.0 cm film was fixed on the support using double-sided adhesive, at an angle of 90° to the film surface which allowed the observation of the cross section of the film. All the film samples were cut with a new razor blade to limit morphological damage. Films were focused up to 15000×, and magnifications ranging from 800x to 8000x were selected, with an intensity of 8 kV and absolute pressure of 230 Pa in presence of water (RH ~30% at 5 °C). No special preparation, such as palladium or gold coating, was necessary for ESEM observation.Film structure was observed using the light microscope  with coupled camera . The uncoated plum skin, the skin of plums immediately after dipping, and the skin of plums coated and dried were focused up to 400x, and magnifications ranging from 10× to 40× were selected. No special preparation was necessary for microscopic observation.Film microstructure was also observed using a polarizing microscope BX51 . Therefore, small sections of plum skin (approximately 1.0 × 0.5 cm) were cut off with a sharp razor blade. One untreated skin sample was observed directly and two other samples were coated by dipping the skin into a solution of starch film-forming solution. Coated plum skin was observed directly and after air drying. For light microscopic analysis, the samples were placed on microscopic slides and illuminated from the top side with an external light source (100 Watt gooseneck lamp). For epifluorescence images, an Olympus U-MBFL3 reflected light brightfield mirror cube equipped with a built-in neutral density filter was used. Excitation was performed with an Exfo X-Cite 120Q lamp. Images were taken at magnifications of 1000×, 2000× and 4000×.2 m−1, from 650 × 102 to 4000 × 102 m−1, and determined at 25 °C on films equilibrated at 53% RH. This analysis aimed at determining the modifications at the molecular scale of the surface induced by the different coating component addition.The Fourier transform infrared spectra from each film were obtained using a spectrometer IFS 28  by means of attenuated total reflectance (ATR) using ZnSe crystal. Samples were cut from self-standing films by shape razor and scanned. All the spectra had an average of 64 scans at a resolution of 4 × 10All experiments were prepared using completely randomized designs. Statistical analysis was performed with Statgraphics Plus, version 5.0 . The analysis of variance (ANOVA) and Fisher’s least significant difference (LSD) multiple comparisons were performed to detect significant differences in the properties of the films. The significance level used was 0.05.The natural wax structures existing on the plum surface can act as a component of the starch-based coating. By alcohol groups, epicuticular crystal wax merges with the components of the starch–glycerol film-forming solution. Hence, starch and starch–whey protein coatings have the same properties as starch–oil films and probably starch–whey protein (80/20)–oil systems. ESEM and light microscopy confirmed that layered structures were combined. The epicuticular wax crystal coating was not visible few hours after coating, but still exists, dispersed in the starch–glycerol–water system. As such, a 3-layer structure did not appear on the surface of the fruit as expected, but a thin (~28μm) emulsion was evident instead. Resistance to oxygen and water vapor permeability was improved. To summarize, observation and quantitative data illustrate a positive relationship between the presence of a starch or starch–whey protein coating and the shelf life of plums.Further studies should focus on the key problem of how these findings can be used to extend the shelf life of the fruits. The weightings of the individual coating components have to be varied so that a maximum keeping quality of the fruits can be ascertained. In addition, research has to take into account several additional aspects such as consumer acceptability and costs of the coating material.


FMR1 gene in the normal population is unknown. In this study, we investigated the frequent of AGG loss, in one or two alleles within the normal population. The role of AGG in the FMR1 stability has been assessed by correlating AGG loss to the prevalence of premutation/full mutation in two ethnic groups differing in their consanguinity rate: high versus low consanguinity rate (HCR vs. LCR).The prevalence and the role of AGG interruptions within the The CGG repeat allele size and AGG presence were measured in 6,865 and 6,204 females belonging to the LCR (5%) and HCR (>45%) groups, respectively, by Tripled‐Primed‐PCR technique.A lower prevalence of the premutation was observed in the HCR (1:158) as compared to the LCR group (1:128). No full mutation was found in the HCR females while in the LCR group the prevalence found was 1:1,149. Homozygosity rate was higher in the HCR population compared to the LCR group.The overall AGG loss was higher in the HCR population than in the LCR and increased with increased CGG repeat number in both ethnic groups.Although we observed a significantly higher rate of homozygosity and AGG loss in the HCR group, this did not affect the prevalence of the premutation and full mutation in this population. Their prevalence was significantly lower than in the LCR population. Finally, we discuss whether the loss of AGG could be also a polymorphic event but not only a stabilizing factor. To further study the role of the AGG interruptions in the 22.1More than 850,000 habitants are living in the Negev region, among them, are the Bedouins which are represented by approximately 250,000 habitants  and Low Rate Consanguinity , respectively. The pattern of AGG loss was assessed in 12,769  of the 17,087 tested females.We also measured the CGG repeat allele size in 323 males that were admitted to our laboratory for Fragile X DNA testing. Of them 191 males belonging to the LCR group and of the 132 males belonging to the HCR group, 120 and 98 males were also tested for the loss of AGG interruptions. Written Informed consents were obtained from all the participants in this study.2.2We defined the index of the patterns of loss of AGG Figure  as follo11‐one AGG loss in the 1st position in one allele.12‐one AGG loss in 2nd position in one allele.111‐one AGG loss in 1st and 2nd position in one allele.1122‐AGG loss in one allele in the 1st position and in both alleles in the 2nd position  or QIAsymphony DNA Midi Kit (96) ‐931255 and the MagNa Pure or QIAsymphony machine according to the manufacturer's instructions.2.4FMR1 PCR assay  as previously described  and electropherograms were analyzed using GeneMapper 4.0 (4.1 for 3500xL data)  was amplified with the Amplidex 2.5FMR1 gene. Two of them, DXS548 and FRAXAC1 were microsatellite markers and were genotyped and visualized using capillary gel electrophoresis. In addition, the two SNPs downstream of the CGG repeat element (rs25714 and rs4949) were analyzed using Taqman SNP genotyping following the manufacturer's protocols. Detailed method described by , rs4949 (ATL1) located proximally and distally to the CGG repeats region of the  Tassone ).2.6p‐values less than .05 were considered statistically significant. pz was defined in this study as the probability of Z‐test and pchi as the probability of the Chi‐squared test. Correlations between the length of CGG repeats and the AGG loss for the different CGG categories were determined by Pearson correlation and by Spearman's Rho. r‐value of 1 by both correlation tests was considered as a positive perfect correlation and p‐values less than .05 were considered statistically significant.Statistical analysis was performed using SPSS. Chi‐squared test was used to assess the association between consanguinity and the loss of AGG sequences. Z‐ Score Calculations for 2 Population Proportions were used in order to determine whether the two groups differed significantly on some single  characteristic. 33.1n = 35) belonged to the 58–69 CGG repeat category, 9 females to the 70–89 CGG repeat category and only 6 females carried an allele greater than 90 CGG repeats. The prevalence of the premutation in this population was 1 in 158. None of them had the full mutation. The LCR population included 9,114 females and 99.1% of them were in the ≤ 57 CGG repeat length category and 72 females (0.78%) in the 58–199 CGG repeat category. The prevalence of the premutation in the LCR population was 1 in 128. Eight females had the full mutation (>200 CGG repeats) and hence, the full mutation prevalence was 1 in 1,149. The highest CGG repeat allele length prevalence in both populations was in the 28–32 CGG range  females were in the ≤ 57 CGG repeat length category and 50 (0.6%) in the 58–199 CGG repeat premutation category. Most of the females .All patterns of AGG loss in the heterozygous subcategories  in the HCR and LCR groups show that: (a) in both groups 28–32 CGG repeat length is the highest prevalent ; (b) a statistically significant higher rate of AGG loss  was observed in the HCR compared to the LCR group in the total CGG length repeat  were obtained for both HCR and LCR‐heterozygous subjects indicating that their statistic distributions are nonparametric.A linear correlation between the overall AGG loss which increases as the CGG repeat increases is shown in Table 3.5pchi = .000039, pz < .05).The different patterns of AGG loss in one allele  and in two alleles  in each ethnic group are shown in Table n = 265 and n = 183) was low we could not show statistical significance, yet, significant difference was observed in the pattern of AGG loss 21 in the HCR compared to LCR group in the 33–40 CGG repeat range .The patterns of AGG loss in both alleles revealed that in general, their prevalence in both ethnic groups was low: 11.2% versus 8.4% in HCR and LCR group. Since the number of subjects in each category . Within the 132 males belonging to the HCR group two had a premutation allele (60 and 67 CGG repeats). The AGG loss, determined for 98 males, was almost twofold higher in the HCR than in the LCR group (29% vs. 15%).3.7During our routine testing for FXS that occurred between 2016 and 2017, we found 16 cases of 5,994, with alleles of three different CGG repeat sizes and eight cases showed AGG loss. Specifically, the 111 patterns of AGG loss were observed in three cases,while, the 11 patterns of AGG loss were observed in five cases.3.8pz < .0001 in the HCR compared to the LCR group.No significant differences were observed between the HCR and LCR groups in the haplotype analysis performed on 147 and 144 samples from LCR and HCR groups, respectively. However the level of homozygosity was significantly higher , we observed the presence of an extra allele within the normal range in 0.27% of the cases . An AGG loss was found only in half (n = 8) of our cases. Another question regards the role of AGG on the stability of the FMR1 allele which was reported to be limited only up to approximately 90 CGG repeats, while, beyond this point no stabilization effect is observed . We expected an increase rate of AGG loss that according to the AGG stability theory should have increased the prevalence of the premutation/full mutation prevalence. However, in the HCR population, the prevalence of the premutation was 1 in 158 and no full mutation was detected. Specifically, 0.6% females carried an allele in the premutation range (58–199 CGG repeat) in 12% greater than 90 CGG repeats. In comparison the prevalence of the premutation in the LCR population was 1 in 128:0.9% of the females carried an allele > 57 CGG repeats and among them 11.5% were above 200 CGG repeats, with a the full mutation prevalence of 1 in 1,149.Inter marriage (consanguinity) within families decreases the genomic variability and increases the homozygosity rate. In our study, we looked at the role of AGG in the instability of the The premutation prevalence in Israel according to the Ministry of Health as described by Zlotogora et al.  for 44,5This is the first study describing consanguinity as related to the premutation prevalence in Israel. Compared to the different published prevalence it is the lower prevalence found in the HCR population in the Negev Region in Israel.The prevalence of all patterns of AGG loss observed was approximately 35%, which is much higher than that published by Weiss et al. . In theipchi = .00001, pz = .001) , followed by the pattern 111  and then by pattern 12 . However, this profile changed beyond 32 CGG repeat length, where, the most prevalent AGG pattern loss was 111 followed by pattern 11 and then by pattern 12 .The patterns of AGG loss 21, 211, and 22 are the result of mating of individuals carrying each one the same pattern of AGG loss. Indeed, we found a statistical significant association between these patterns of AGG loss and consanguinity .The loss of AGG was 4.8% higher in the HRC group compared to the LRC group. Although the association between consanguinity and homozygosity was expected, the low prevalence of the full mutation in the HCR population  is not in agreement with the theory of the AGG loss as an important factor in the instability of the FMR1 gene.We found that the homozygosity, as well as, the rate of AGG loss were statistically significantly higher in the HCR as compared to the LCR population . Finally, Weiss et al. (FMR1 gene.In summary, as expected, our results demonstrate that consanguinity affects the homozygosity as well as the prevalence of AGG loss. However, it did not affect the prevalence of the premutation and full mutation of the nd Kamal  aimed tos et al.  showed nFMR1 gene. Although we did not find difference in the haplotype analysis between the two groups, the involvement of an ethnic distinct stabilization factor could still play an important role. Our results also show that there might be a tendency in the pattern and rate of AGG loss positively correlated to the CGG repeat length.According to our results it could be suggested that the loss of AGG is polymorphic phenomenon in the general population that play also a role in the stability of the CGG repeat length in the FMR1 instability, which is, to date, still not fully understood.Finally, further studies are warranted to clarify these results as well as the mechanism of There is no author in this paper with conflict of interest.


External perturbation forces may compromise standing balance. The nervous system can intervene only after a delay greater than 100 ms, during which the body falls freely. With ageing, sensorimotor delays are prolonged, posing a critical threat to balance. We study a generic model of stabilisation with neural delays to understand how the organism should adapt to challenging balance conditions. The model suggests that ankle stiffness should be increased in anticipation of perturbations, for example by muscle co-contraction, so as to slow down body fall during the neural response delay. Increased ankle muscle co-contraction is indeed observed in young adults when standing in challenging balance conditions, and in older relative to young adults during normal stance. In parallel, the analysis of the model shows that increases in either stiffness or neural delay must be coordinated with decreases in spinal sensorimotor gains, otherwise the feedback itself becomes destabilizing. Accordingly, a decrease in spinal feedback is observed in challenging conditions, and with age-related increases in neural delay. These observations have been previously interpreted as indicating an increased reliance on cortical rather than spinal control of balance, despite the fact that cortical responses have a longer latency. Our analysis challenges this interpretation by showing that these observations are consistent with a functional coadaptation of spinal feedback gains to functional changes in stiffness and neural delay. Being able to stand still can be difficult when faced with an unexpected push. It takes the nervous system more than a tenth of a second to respond to such a perturbation, and during this delay the body falls under the influence of its own weight. By co-contracting their ankle muscles in anticipation of a perturbation, subjects can increase their ankle stiffness, which slows down their fall during the neural delay. Young subjects indeed adopt this strategy when they need to remain particularly still (for example when they stand in front of a cliff). Older subjects adopt this strategy even during normal standing. We present a model of standing balance that shows that this postural strategy provides partial compensation for the increase in neural delays with ageing. According to our model, increasing ankle stiffness only improves balance if it is accompanied by a decrease in sensorimotor gain. This provides a novel and functional interpretation for the decrease in spinal feedback observed during ageing, and observed in young subjects when they stand in challenging balance conditions. External perturbation forces may compromise standing balance . CurrentThe difficulty with this approach is that the neural feedback control of movement introduces delays. The shortest spinal delay after which a change in contraction can be observed following a direct electrical stimulation of the sensory nerve as in  is arounDuring this neural response delay after a perturbation, the movement of the body is entirely determined by the body and environmental mechanical properties, such as stiffness, inertia and weight. Ankle stiffness can be actively increased through co-contraction of the ankle muscles . Young sTo understand the relative contributions of ankle stiffness, neural delay and sensorimotor gain to balance performance, we analyze a widely used model of standing balance. We propose a novel method to determine how sensorimotor gains should adjust to changes in ankle stiffness and neural delay. We show that the most effective strategy to improve balance relies on the co-adaptation of ankle stiffness and sensorimotor gains. Increasing ankle stiffness in advance of a perturbation can improve robustness to perturbations, by reducing the amplification of perturbations during the neural feedback delay. However, this improves balance only if the increase in ankle stiffness is coordinated with a decrease in sensorimotor gain, to prevent over-compensation. This strategy is also the most effective for maintaining balance performance despite increasing neural delay.The decreased spinal feedback observed during normal standing in older subjects, and during standing in challenging balance conditions in young subjects, should therefore not be interpreted as the mark of a more cortical control of balance, but on the contrary as the sign that motor stabilisation is achieved through an increased reliance on ankle stiffness.τdelay = 0.14 s, the body centre of mass is accelerated by gravity. We model the body mechanics using the single inverted pendulum model, which is widely used to model human stance in the sagittal plane [Θ0 (detailed in the Methods), the ankle angle θ follows:After a perturbation of stance, the earliest change in muscular contraction occurs with a delay of around 100 ms in young subjects . The change in force due to this muscle contraction is only observed after an additional 40 ms . During al plane ,21. Afteτmech, which captures the effects of the rotational inertia around the ankles J, ankle stiffness k and weight mg. Any increase in ankle stiffness k up to the critical ankle stiffness Kcrit = mgLcos(Θ0) increases the mechanical time constant of the body. Ankle stiffness thus slows down falling during the response delay [S.The dynamics are thus governed by the mechanical time constant se delay . During We wish to determine how the neural feedback control of balance is affected by an increase in ankle stiffness, with and without an increase in neural delay. For this, we first assume a simple model for the control of standing balance, and show that both strategies require a decrease in sensorimotor gain. We then show that these results generalise to more complex neuro-mechanical models.We first assume that balance is achieved through delayed feedback control of position and speed. Such a control strategy has previously been suggested to model human balance in both the sagittal  and lateτmech) and neural response delay, there is a limited range of proportional gains p and derivative gains d that can stabilize the system [For any combination of mechanical parameters . This suggests that when subjects increase ankle stiffness to compensate for an increase in neural delay, they must decrease their sensorimotor gains.With aging, there is in increase in feedback delay . According to our model, with increasing feedback delay , the rantiffness , black aWithin the range of stable feedback, large gain and low damping lead to oscillations, whereas low gain and large damping result in slow compensation for perturbations (as illustrated in the Methods). To determine how sensorimotor gains are affected by an increase in stiffness or neural delay, we first determine the best combination of feedback gains and the resulting balance performance, for a given stiffness and delay.ω0, the fastest compensation without oscillations occurs for the critical damping ζ = 1. For such critical damping, the characteristic equation has a unique double root −ω0. If a perturbation brings the system away from its equilibrium position, then the system is returned to its initial position following the time-course We assume that the best combination of feedback gains results in fast compensation of perturbations without oscillations. Subjects typically adopt such feedback gains when responding to lateral perturbations of stance . In secoλ0. The characteristic time to return to equilibrium is then τbalance , and the corresponding critical feedback gains. We then consider that the delay is doubled  responses of subjects to visual and platform rotation perturbations, Peterka included integral feedback in their model of standing balance . We applFinally, standing balance in the sagittal plane is better modelled by considering not only ankle angle but also hip angle  and kneeWhen a subject is asked to stand in a situation in which it is particularly important not to move , a decrecrit and corresponding critical gains, indicated as black dots in However, our modelling results show that a decrease in spinal gains alone is not beneficial for supra-spinal control. Cortical and other supra-spinal contributions to balance have longer delays than the spinal stretch reflex. During this supra-spinal delay, the dynamics are determined only by the body mechanics and the spinal control. A decrease in spinal gains (without a corresponding increase in stiffness) causes the CoM to fall faster and further during this delay period . Our modelling results generalise to multi-dimensional systems and therefore predict that standing balance may also benefit from the stiffness of muscles acting at joints other than the ankle, if they slow down the amplification of perturbations during the response delay. This has indeed been shown by De Groote and colleagues, who asked human subjects to stand still despite external perturbations . They obWith aging, there is an increase in the H-reflex latency of the ankle muscle soleus by 10 to 15% . The rescrit. The increase in ankle stiffness with aging, when measured with relaxed muscles may be more than 20% [crit increase in ankle stiffness required by the co-adaptation strategy is achieved during normal ageing.The co-adaptation strategy requires a 30.6% decrease in proportional gain. This is consistent with the 30 to 36% decrease in soleus H-reflex occurring during aging –9. The mthan 20%  , but rather an adaptation for increased delays.We have presented a generic model of stabilisation with neural response delays, which highlights the importance of co-adapting ankle stiffness and sensorimotor gains. In situations in which it is particularly important to remain immobile , our model suggests that the most effective way to improve balance is to increase ankle stiffness and decrease sensorimotor gains. Indeed, the increase in ankle stiffness slows down the CoM fall during the neural response delay. As the CoM falls more slowly, a smaller increase in contraction is then required to return it to its initial position. This co-adaptation strategy allows perturbations to be cancelled faster, and with smaller excursions in both the CoM and CoP. We have reviewed experimental evidence that, in challenging balance conditions, young subjects indeed decrease spinal gains and at least partially increase ankle stiffness.During aging, the neural response delay to perturbations of stance increases. According to our modelling results, an increase in delay without adaptation of either stiffness or gains results in oscillations of the CoM, as well as increased excursions of both the CoM and CoP. To prevent such oscillations, the sensorimotor gains must be decreased. This gain adaptation strategy prevents CoM oscillations, and decreases CoP excursions, but further increases the CoM excursions. According to our model, the most effective way to maintain balance performance with increasing neural delay is to increase stiffness and decrease gains. This co-adaptation strategy prevents the increase in CoM excursions, and further decreases the CoP excursions. We have reviewed experimental evidence that in quiet standing, the spinal gains of older subjects are decreased relative to young subjects, and their ankle stiffness is increased.Interestingly, the co-adaptation strategy not only reduces the CoM excursions (or prevents their amplification with increasing delays), it also reduces the CoP excursions. In the simulations we have presented, the CoP range is assumed to be unlimited. In human stance however, the position of CoP is limited to the extent of the foot. By reducing the CoP excursions after a perturbation of a given amplitude, the co-adaptation strategy allows larger perturbations to be compensated for without requiring unrealistic torques. This may be particularly important for older subjects who have a reduced range of CoP motion ,33. In pThe decrease in spinal gains observed when standing in challenging balance conditions, or during aging, is commonly considered as the sign of an increased cortical control of balance. However, our modelling results show that decreasing spinal gains (without co-adapting ankle stiffness) does not provide any advantage for the cortical control of stance in challenging balance conditions. As supra-spinal control has longer delays than spinal control, if the spinal gain is decreased, then the CoM falls further and picks up more speed by the time the supra-spinal control intervenes. This increases the difficulty of the task for the supra-spinal control. The co-adaptation strategy on the contrary reduces the CoM excursions. Thus, independently of supra-spinal control, the task of standing balance becomes easier if spinal gains are decreased in combination with an increase in stiffness. We have also shown that a decrease in spinal gains is an effective strategy to compensate for age-related increases in delays, independently of any assumptions about supra-spinal control. According to our model, the most effective strategy is again a combined increase in stiffness and decrease in gains. We therefore argue that the reduction in stretch reflex observed during aging and in challenging balance conditions is not the hallmark of increased cortical control, but rather reflects an increased reliance on ankle stiffness.Brainstem- or cortically-mediated visual and vestibular responses to perturbations of stance also play an important role for maintaining balance ,36. ThesPrevious modelling studies have determined the stability regions for the feedback gains of the delayed PD controller  to quiet standing sway measures in young and older subjects . They acThe critical sensorimotor gains are close to the minimal gains required for stability ). If a sThis might explain the recent experimental observations that spinal gains are actually increased rather than decreased when standing at a very large height. When subjects stand at the edge of an elevated platform of 1.6 m height, the H-reflex is decreased (4). The longer latency (120–220 ms) response to a rotation of the ankle joint is however increased . MoreoveWhen standing normally, it might also be the case that young subjects adopt robust gains which are larger than the critical gains. In this case, the decrease in H-reflex observed in challenging balance conditions may reflect the subject’s attempt to adopt gains which are closer to the critical gains, thus improving the immobilisation of the CoM.The relation between H-reflex and muscle contraction has been extensively studied in a different experimental paradigm . In thisAnkle stiffness during stance is measured by imposing a rotation of the ankle and measuring the immediate change in ground reaction torque that ensues. Experimental measures vary from 40% to 90% of the critical stiffness ,35,44,45When standing in normal conditions, young subjects typically do not co-contract their ankle muscles; however, they are able to do so when it is particularly important to remain immobile ,6,19. LiWhen measured with relaxed muscles, ankle stiffness is larger in older adults relative to young subjects . AdditiomgL sin(Θ) . To understand the relative roles of body mechanical properties and feedback control in stabilisation, we decompose the ground reaction torque into a mechanical component K(Θ) due to ankle stiffness, which changes immediately upon a change in ankle angle, and a component C due to feedback muscle contraction with only changes after a delay τdelay after a mechanical perturbation. The sum of the external torques affects the person’s rotational momentum We consider the widely used model of human stance in the sagittal plane: the single inverted pendulum model presented in Θ0 with muscular contraction C0, and linearize around this equilibrium, introducing θ, k, c such that:We consider that the person is initially at equilibrium at an angle After linearization, the change in rotational momentum becomes:The dynamics thus follows:τmech corresponds to k = 0. With mechanical parameters corresponding to human stance ≈1), the minimal mechanical time constant is thus:The lower bound of τd,0 = 0.14 s.For the neural feedback control, we consider a minimal time delay θ and speed θ(t = 0) = θ0, and Suppose an initial perturbation shifts the CoM position t = τdelay, the initial perturbation amplitude θ0 has been amplified to the value Thus, by the time the nervous system intervenes at We consider that the neural response follows a delayed proportional-derivative feedback controller, such that the dynamics of the system follows:The characteristic equation of the system is therefore:ω>0:The stability boundaries for this equation have been previously published . A derivx = ωτdelay, D = dτdelay, x>0:We introduce the dimensionless variables: S. For a given S, the stability limits for the proportional gain p scale with d scale with τdelay. We therefore plot the stability limits in terms of the dimensionless gains D = d/τdelay. For each value of S, there is a limited range of feedback gains  that can stabilize the system. This range is larger for slower values of S, suggesting slower systems are more robust to an inappropriate calibration of the feedback gains. Additionally, with increasing S, the range of stable gains shifts towards higher values. This shows that if the system speed S changes, then the feedback gains must be adjusted to maintain stability.In The characteristic equation of the linearized inverted pendulum with delayed feedback control is:X = τdelayλ. The roots of the characteristic equation are given by:We introduce the dimensionless variable Critical damping is defined for second order systems governed by a characteristic equation of the form:To define critical damping for this system, we first introduce a rational function approximation for the delay, first proposed by Pade . With thThe first order Pade approximation of the delay is given by:A dynamical systems interpretation of this approximation is provided in The roots of the approximate characteristic equation are thus the roots of:ω0, the fastest compensation without oscillations occurs for the critical damping ζ = 1. For such critical damping, the characteristic equation has a unique double root −ω0. Higher damping results in slower compensation for perturbations, whereas lower damping results in oscillations. We generalize the notion of ‘critical damping’, and consider that critical feedback in an nth order system corresponds to the system having a unique negative root −ω0 of order n.In second order systems governed by a characteristic equation ω0.We therefore consider that our system is at criticality when is has a unique triple negative root −The coefficients of the characteristic equation must therefore correspond to the coefficients of the polynomial:crit, Dcrit for a given relative speed S, we solve for  the system of equations:To find the critical feedback gains Pω0 as a function of S by removing  from the equations:First we determine crit = 2−3ω0According to (3): Dcrit = 2D−S2−3ω02 = 4−6ω0−S2−3 ω02According to (2): PReplacing in (1):ω0+2)3 = 4(4−S2)>0Thus: (ω0+2), and two complex conjugate solutions. We take the real solution:This equation admits one real positive solution for (Replacing in (3): Replacing in (2): τdelay = 0.14 s with critical stiffness  the system of equations:For a given speed The resulting time to recover balance and critical gains as a function of S are shown in A such that:We consider that the system has acceleration feedback A, we must use the second order Pade approximation:To calculate the critical feedback gains with acceleration feedback With this approximation, the roots of the characteristic equation are given by:With critical feedback, this should have the same roots as:We numerically solve the system of equations:The resulting time to recover balance and critical gains as a function of S are shown in i such that:We consider that the system has integral feedback Introducing the dimensionless variable Using the first order Pade approximation, the characteristic equation becomes:With critical feedback, this should have the same roots as:We numerically solve the system of equations:θ and delayed feedback control C, whose dynamics are governed by:We consider an N-dimensional dynamical system with state ST of the dynamics matrix S is diagonalizable, and introduce the basis set  of eigenvectors of ST and their corresponding eigenvalues , such that for every i:We consider the generic case, in which the transpose θ into α, such that, for every i, αi is the dot product of the vectors ei and θ:We use this basis set to perform a transformation of coordinates of the state αi follows the dynamical equation:Each component The dynamics is thus decomposed into a set of N components, each of which follows a single-dimensional dynamics for which the analysis presented in the previous sections holds.τi such that When considering a model of human stance with several joints , the dynamics can be decomposed into such a set of components. Each of these components is a linear combination of the various angles, and its dynamics is governed by a mechanical time constant S1 Text(DOCX)Click here for additional data file.S2 Text(DOCX)Click here for additional data file.


Balkan Medical Journal, the official university/academic journal of Trakya University Faculty of Medicine, is under the ownership of the deanery of the faculty and aims to propagate the current medical scientific knowledge in the Balkan region. The rectorate of Trakya University logistically supports the journal. On the other hand, the editor-in-chief is scientifically independent and cannot be pressurized or manipulated in any way regarding the scientific contents and operation of the journal. Nonetheless, the editor-in-chief has obligations toward the journal owner and editorial board. His/her editorial independence, rights, and responsibilities are aptly described and documented by the international scientific publishing communities. These include the editor’s responsibilities, autonomy, and accountability of the journal’s scientific content ,2,3,4,5.Since its establishment, the Balkan Medical Journal has aimed to select and publish distinctive, original articles in general medicine to contribute to universal scientific knowledge and to share its experience through educational activities for authors, reviewers, and editors . DespiteA careful review of the literature reveals that the Balkan region has taken permanent steps towards institutionalization with the successful publication practices of journals. Croatian Medical Journal stands out as the most significant journal in this regard. A paper by Marusic et al.  in 2005 For a journal to be successful in the international arena, reasonably organized managerial processes are required. Clear definitions of work packages, workflows, duties and responsibilities, and assignment and dismissal criteria are essential for the transparency of managerial processes. The journal’s owner, editor-in-chief, editorial board, publisher, and authors effectively communicating with each other within the framework of rules will prevent chaotic and destructive conflicts that may harm the reputation and its mission. Furthermore, the inscription of such rules promotes scientific autonomy of the editor and prevents misuse of editorial freedom thereby resulting in the continuous success of a journal.The efforts to establish a legal regulation, an agreement between the owner and the editor, and an ombudsman began approximately five years ago for Balkan Medical Journal. A draft text of legal regulation was submitted for discussion to the editorial board at that time. Intensive efforts have been carried out to achieve a certain degree of maturity. Since no other university/academic general medicine journal in Turkey is also indexed in internationally published SCI-E and Medline, it took time for all stakeholders to draft a consensus document. Thanks to the previous editor-in-chief of Balkan Medical Journal and the current vice-rector of Trakya University responsible for scientific journals (Cem Uzun), who has encouraged all editors of Trakya University journal to establish legal regulations, we have a transparent scientific publishing atmosphere to discuss, write, and own individual legal regulation for each journal. Throughout this process, the journal’s management has endeavored to consult the opinions of the journal’s owner and the university legal regulation committee to establish a legal regulation for the journal that is ethical, practical, sustainable, and conforms to the national laws and international scientific values. We believe the journal legal regulation, finalized in January 2019 and presented below, serves as a role model for all scientific circles and the Balkans and will contribute to the advancement of institutional scientific publications of medicine in the region. Nonetheless, this legal regulation is open to being updated and improved whenever needed, as our other policies. The journal’s editorial board extends its gratitude to all current and past employees, the Dean’s Office of the Faculty of Medicine of Trakya University, and to the current rectorate of Trakya University for their valuable contributions and encouragement to building the development of the journal legal regulation to protect editorial freedom and provide successful journal management of all Trakya University scientific journals.


The aim of this paper was to produce a new composite material based on carbon and iron oxides, starting from soluble starch and ferric chloride. The composite material was synthesized by simple thermal decomposition of a reaction mass obtained from starch and iron chloride, in an inert atmosphere. Starch used as a carbon source also efficiently stabilizes the iron oxides particles obtained during the thermal decomposition. The reaction mass used for the thermal decomposition was obtained by simultaneously mixing the carbon and iron oxide precursors, without addition of any precipitation agent. The proper composite material can be obtained by rigorously adhering to the stirring time, temperature, and water quantity used during the preparation of the reaction mass, as well as the thermal regime and the controlled atmosphere used during the thermal decomposition. Synthesized materials were characterized using thermogravimetric analysis, X-Ray Diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infra-red spectroscopy (FT-IR). The performances of the obtained material were highlighted by studying their adsorbent properties and by determining the maximum adsorption capacity for arsenic removal from aqueous solutions. Iron oxides are widely used in chemical synthesis, in the wastewater remediation process, as well as in the treatment of drinking water. Beside these examples, iron oxides are used as catalysts in various industrial processes ,2, as adHowever, the actual usage of such small particles is really difficult because it is a hard job to separate such small particles from the reaction environment. Therefore, an alternative of this would be to attach such nanoparticles to different macro-sized supports, which must be chemically, mechanically and thermally stable. One of the most utilized supports in real application is represented by carbon, in the form of graphite or activated carbon, which presents a large surface area, as well as being a relatively cheap and environmentally friendly material. A green way to produce such a support material can be represented by the pyrolysis of various organic compounds with large molecular size, such as cellulose, starch, and bio-polymers . The activity of such material can be increased by attaching iron oxide particles onto the support material, a complex process which can be accomplished in several stages. Such a process has a relatively high resources consumption, and at the same time it is difficult to implement physical–chemical methods ,14,15.Surprisingly until the 2000s no natural polymer had been considered for the production of carbon based support materials . Since tSimultaneously, the thermal decomposition temperature controls the particle size and stability, as well as the pore size and volume. One other factor which controls the pores form and volume is represented by the natural ability of amylose and amylopectin chains from natural polymers (e.g.starch) to assemble in a supramolecular lamellar structure with nanometric dimensions, presenting crystalline and amorphous areas . At the Zhang et al.  and LianAmylose is a natural polymer with a molecular mass between 10,000 and 340,000 Daltons and is formed from maltose units linked by 1-4 α glyosidic bonds, while having a linear structure, with a spiral form. Each spiral consists of six fragments of glucopyranosyl groups. Amylose is a compound relatively soluble in cold water, which does not form a sticky dough with hot water. d-glucopyranose linked by 1-4 and 1-6 α-glycosydic bonds. This compound is not soluble in cold water, but can be dissolved in hot water which is responsible for dough formation [The structural units of amylopectin are maltose and isomaltose having a branched structure, an amylopectin chain is formed from α-ormation .In acidic environment hydrolysis of soluble starch occurs by breakage of the glycosidic bonds forming glucose molecules. Experimental data confirm that the acidic hydrolysis of starch takes place preferentially in amorphous areas, areas rich in amylose, while areas rich in amylopectin are less susceptible to hydrolysis .3), sodium trimethyl phosphate, epichlorohydrin, acetic acid, adipic acid [Starch can be modified by using acidic media with slow boiling, forming oxidized starch, crosslinked starch, partially esterified or etherified starch, or even starch converted to cationic derivatives. Production of new bonds occurs at the surface of the starch dough, which reacts with bi- or poly-functional specific reagents like phosphorus oxychloride , followed by the thermal decomposition in a nitrogen atmosphere (second stage). Employing such a synthesis route, the production of iron oxide particles is then possible without adding any precipitation agents. After production, the new composite material was characterized and tested as an adsorbent for arsenic removal from aqueous solutions.Composite material was synthesized as described in Production of the desired composite material using starch and iron chloride as precursors can be understood if we consider that by dissolution of anhydrous iron chloride in water a hydrolysis process takes place.By simple mixing of starch with water a starch dough is produced without any changes in the starch structure. By dissolving anhydrous iron chloride in water, iron (III) cations and chloride anions are produced, with a concomitant decrease of the solution pH.Hydrolysis behavior of iron (III) cations in diluted solutions at low pH values leads to the formation of iron oxy-hydroxides and iron hydroxides according to the mechanism below ,33:Fe[) ,22.Fe[, this speed ramp also avoided destruction of the carbon structure.Based on data obtained from the analyzed thermogravimetric curves it was established that the thermal treatment of “starch–iron chloride” paste must be carried out in a controlled nitrogen atmosphere using a temperature of 400 °C. These conditions were chosen in order to preserve the carbon structure of the obtained material. Experimental data confirm that iron oxide was obtained when the sample was heated by using a relatively low speed ramp ; the presence of some peaks specific for Fe–O bonds, peaks are located in the interval 1000–450 cm−1; an increase of the C–C, C–O and Fe–O specific bands is located between 2000 and 1000 cm−1; and that by comparing the FT-IR spectra of Am-Fe material with the spectra recorded for pure starch, we notice that the starch IR vibrations are shifted towards higher wave numbers having lower intensities [From the data depicted in ensities .Recorded FT-IR spectra confirm that thermal decomposition of Am-Fe material leads to the formation of a carbon structure which contains iron oxide particles bonded onto the carbon surface by oxygen bonds.The recorded XRD spectra obtained for three synthesized materials are depicted in 3O4 particles. By looking at these spectra, we can also conclude that the stirring time has no effect over the species that appear in the final product.From spectra depicted in Recorded SEM micrographs are depicted in Based on data presented in the SEM pictures we can observe that the magnetite particles have a relative uniform size. From Data obtained for sample 3, which was obtained using a low stirring time (20 min) but with the highest decomposition temperature (58 °C), confirm that a heterogeneous material containing few magnetite particles with higher size distribution was produced. Increasing the stirring time at this temperature has no beneficial result due to the occurrence of gelatinization which favors the separation of iron oxides from the starch particles, leading to partial separation during the paste drying.An important application of the newly synthesized materials is represented by their usage as adsorbents for removal of different pollutants the high affinity of arsenic ions for iron being well known ,43,44,45Taking into account the procedure, from each sample a quantity of 0.1 g was measured, which was mixed with a set of 25 mL solutions containing variable arsenic concentrations. The adsorbent material was kept in contact with the arsenic solution for one hour in a thermostatic shaker bath at 200 rotations per minute. After one hour, the solution was filtered and the residual arsenic concentration was measured in the solution obtained after the adsorption process was performed.The maximum adsorption capacity obtained from the experimental studies is presented in On analyzing data presented in When a magnetite content decrease was observed then the maximum adsorption capacity of the newly produced adsorbent material decreased. In the case of sample S3, the maximum adsorption capacity was 26 μg As(V) per gram of adsorbent, which is seven times lower than the maximum adsorption capacity obtained for sample S2. Also, based on data presented in The composite material was synthesized using the following route: 20 g of soluble starch (Merck—designated as Am) was mixed with 25 mL of distilled water, in triplicate). Each obtained sample was heated at a different temperature, 45, 55, and 58 °C. When the starch mass attained the desired temperature, over it under continuous stirring a solution of ferric chloride (Merck) was poured with a concentration of 33.33 % (2.5 g of anhydrous ferric chloride was dissolved in 5 mL of distilled water). The reaction mass was stirred continuously for various time intervals  until transformed into a paste. These quantities of precursors were calculated in order to obtain a final ratio of Fe:C equal to 1:10. This ratio was chosen because the experimental data showed that the usage of higher quantities of iron chloride was not recommended because it leads to starch gelatinization, simultaneously it was proved that smaller ratio leads to a composite material with a lower efficiency in various applications .−1, in nitrogen atmosphere), X Ray Diffraction (using a Rigaku Ultima IV XRD diffractometer), scanning electron microscopy (SEM—using a Quanta FEG 250 scanning electron microscope), and Fourier transform infrared spectroscopy (FT-IR—using a Bruker Platinum ATR-QL-Diamond spectrometer).Raw material used for thermal decomposition and the composite material obtained (designed as Am-Fe) were characterized through thermogravimetric analysis , for one hour in a Julabo bath shaker with 200 rotations per minute. The stock solution of arsenic ions was prepared using a standard solution containing 1000 mg As (V) L−1 in 2% nitric. Arsenic residual concentration in solution resulted after the adsorption was determined using mass spectroscopy inductively coupled plasma (ICP-MS Bruker Aurora M90).Synthesized composite material was used as adsorbent material for arsenic removal. So, during experimental work, the adsorbent properties of material were studied by determining the adsorption capacity for arsenic ions. Maximum adsorption capacity was determined by keeping in contact a well measured quantity of adsorbent material (0.1 g) with a volume of 25 mL of solutions with variable arsenic content  L−1) was calculated using the following equation:o—initial concentration of arsenic (V) from solution, (μg L−1); Cf—the residual arsenic concentration (V) from solution, (μg L−1); V—volume of solution, (L); m—mass of adsorbent material, (g).Material adsorption capacity q  particles present a low size distribution, being homogenous, and having a low size.−1, in a nitrogen atmosphere. Such a material can be used as an efficient adsorbent for the removal of arsenic ions from water solutions. The maximum adsorption capacity has a value of 261 μg As(V) per gram, higher then the majority of the adsorption capacities found in the scientific literature.The optimal synthesis parameters are as follows: synthesis temperature 45 °C, stirring time 240 min, thermal decomposition of synthesized paste 400 °C, heating speed rate of 5 °C min


The year 2019 will probably remain a semantic cornerstone in the evolution of the European Society for Gynaecological Endoscopy (ESGE). A new era is beginning in the scientific face of our Society with its new official journal Facts, Views & Vision in ObGyn (FVVO), fulfilling the vision of our pioneers. Since its foundation in 1994, our Society has aimed to encourage the exchange of clinical experience, scientific thought and investigation among European gynaecological endoscopists and practitioners in related techniques.It was obvious from past experience that the official journal of our Society has to provide broad visibility of the published material: our new journal, being open access, is freely accessible to all readers. But not only that; with its 10 years successful history, FVVO succeeded to be included in Pubmed. Thus, the scientific content of the journal is searchable in the main scientific database used by those interested in being informed on a topic of our field.The open access mode and the inclusion in Pubmed are both crucial for the official journal of our Society and ESGE members as the research published in FVVO is visible to the wider scientific community and ultimately will be cited.A clear advantage of the new official journal is that it combines the electronic open access mode with a printed version: this was a longstanding wish of many of our members. The printed version of the journal is really unique as the artist Koen Vanmechelen designs it: the front page is always an art work and selected papers are also covered by special art pieces. Furthermore, the ESGE members and the members of the ESGE Corporate Member Societies will have the opportunity to have the printed version only at the mailing cost.Being an open access journal, it is usual that the authors have to pay for their publication. This is indeed the case for the non-ESGE members. But the ESGE members and the members of the Corporate Member Societies can publish their work, following peer reviewing, free of charge: the Society will cover the expenses of their publication.However, most importantly, our new official journal will be owned by ESGE. This will provide a steady environment for the scientific work in our field and will also give the opportunity to our Corporate Member Societies to be expressed officially by a journal. It is noteworthy that “The Walking Egg” will continue to be present in the journal and that the International Society for Gynaecological Endoscopy (ISGE) has selected FVVO as its official journal.We would like to underline that our intention is to combine the heritage from the successful history of FVVO with the ESGE scientific production and reputation. Therefore, the current Editor in Chief, Prof. Willem Ombelet will continue his successful leadership, supplemented by the ESGE Editor Prof. Ertan Saridogan.We encourage the whole ESGE community to support this new era in the scientific evolution of our Society.ESGE PresidentGrigoris Grimbizis


In this work, the influence of various rolling temperatures and thickness reductions on the dynamic recrystallization (DRX) behavior of AZ31 magnesium alloy sheets was investigated. Meanwhile, the texture variation controlled by DRX behavior was analyzed. Results suggested that, with the help of DRX behavior, reasonable matching of rolling temperature and thickness reduction could effectively refine the grain size and improve the microstructure homogeneity. Using the grain refinement and microstructure homogeneity as the reference, the critical rolling process parameters were 400 °C—30%, 300 °C—30%, and 250 °C—40% in the present work. In terms of basal texture variation, the occurrence of twins produced the largest maximum texture intensity. However, for the sheets with DRX behavior, the maximum texture intensity decreased sharply, but would steadily increase with the growth of DRXed grain. Additionally, for DRXed grains, the <11-20>//RD (RD: rolling direction) grains would gradually annex the <10-10>//RD grains with the growth of DRXed grains, which finally made their texture component become the dominant texture state. However, when the deformation continued, the <10-10> in DRXed grains would rotate toward the RD again. Weighted by the fracture elongation of AZ31 magnesium alloy sheet, the critical thickness reductions were 30–40% under the rolling temperature of 400 °C. Magnesium alloys are recognized as the lightest structure metal and, due to their high specific strength and stiffness, they have received significant attention in many fields, such as electronics, automotive, and aerospace ,2. HowevZ  [Z during plastic deformation. Recently, Liu et al. studied the microstructure of as-rolled AZ31 magnesium alloy sheets with various thickness reductions from 10% to 60%, under similar lower rolling temperature, and pointed out that the 10% sheet consisted of deformed grains, while the 60% sheet completed DRXed grains [Grain refinement was an effective way to improve the room temperature formability of magnesium alloys ,6,7. Manonstant) ,17. Thisd grains . This ded grains . The abod grains ,9,19. Ind grains ,21. For d grains . This fuZ was closely related to the strain rate and rolling temperature. According to the literature [H is the initial thickness, h is the thickness after hot rolling, ν is the rolling speed, and r is the semi-diameter of the roller). Clearly, h was directly affected by thickness reduction. Moreover, H, ν, and r were deemed to be constants in this work. Thus, in the present work, thickness reduction and rolling temperature were the variables which influenced DRX behavior. Based on this, the initial materials were rolled in a single pass with thickness reductions of 20%, 30%, 40%, and 50% under the rolling temperatures of 400 °C, 300 °C, 250 °C, and 200 °C, respectively. Moreover, in order to improve the workability of initial material, all sheets were preheated to 400 °C for 30 min prior hot rolling. Hot rolling was performed on a two-high mill with 110 mm in roll semi-diameter at a roll velocity of 5 m/min. The main rolling parameters, including rolling temperature and thickness reduction, are summarized in Hot-rolled AZ31 magnesium alloy sheets with 4 mm in thickness were used as the initial material in this work. The nominal composition of this initial material was Mg-3wt.% Al-1wt.% Zn-0.2wt.% Mn. As previously described, Zener–Hollomon parameter terature , the rolAs shown in −3 s−1 at room temperature. In this work, the sheets rolled at 400 °C were used as the typical example to investigate the effect of thickness reduction on the mechanical properties of magnesium alloy sheets. Tension samples were cut by wire-cutting from the sheets rolled at 400 °C, along the rolling direction and the transverse direction. Their gauges were 25 mm in length and 6 mm in width. Three samples were each tested for thickness reduction, in order to ensure the repeatability of the results.The samples for microstructure characteristics were cut with the broad surface parallel to the RD–TD plane . After grinding with SiC paper and polishing with 0.05 μm silica suspension, these samples were etched with the use of a solution consisting of picric acid (5.5 g), acetic acid (2 mL), distilled water (10 mL), and ethanol (90 mL). Then, a OLYMPUS GX71 optical microscope (OM)  was applied for the metallographic observations. The mean linear intercept method was used to measure average grain size. However, according to the study by Chen , the relIn terms of the effect of rolling temperature on DRX behavior, it played a similar role to the thickness reduction. Generally, lower deformation temperature led to increased critical deformation for DRX. This meant that DRX only occurred in some regions where the deformation was high enough, while not in other regions if the deformation was below the critical deformation, especially in the core of coarse grains. In view of this, it could be seen in the 200 °C—30% sheet n that thZ [As described above, DRX behavior was a comprehensive outcome of many factors, instead of depending on single factor. Microstructure controlling via DRX behavior was of importance in fabricating magnesium alloy sheets with high performance. This required a reasonable matching for rolling process parameters (thickness reduction and rolling temperature in this work). It is well known that fine grain size and homogeneous microstructure can improve the mechanical properties in magnesium alloys ,28. ThusZ ,30. MoreIn the Orientation Imaging Microscope (OIM) software, the grain average misorientation (GAM) was determined by calculating the misorientation between each neighboring pair of points within the grain . This waIn addition, two points should be noted. First, for the 20% sheets, the fraction of lower GAM value region (blue region) increased significantly with the increase of rolling temperature . Under this thickness reduction, although DRX rarely occurs, dynamic recovery still appeared, which would reduce the dislocation density by dislocation climb, cross-slip, and glide. Moreover, this dynamic recovery generally occurred more at higher deformation temperature. Therefore, the 400 °C—20% sheet exhibited a larger fraction of lower GAM region (blue region). Second, in the sheets with DRX behavior (thickness reduction >30%), the fraction of lower GAM region (DRXed grain) increased with the increase of thickness reduction. This could be attributed to two factors. First, higher thickness reduction produced more nucleation sites, which realized completed DRX more easily; second, higher thickness reduction made new DRXed grains with low dislocation density grow more easily (as described before).As mentioned earlier, under the rolling process parameters of 400 °C—30%, 300 °C—30%, and 250 °C—40%, AZ31 magnesium alloy sheets possessed optimal grain size and microstructure homogeneity. However, for magnesium alloys, their HCP structure was liable to cause strong basal texture, which greatly influenced the mechanical properties ,31. DuriGenerally speaking, the microstructure after hot rolling process could be divided into DRXed grains, subgrains, and deformed grains, and according to many studies, their texture states were different, especially the crystal orientation along the RD ,9. In orIn this work, the DRX behavior of AZ31 magnesium alloy sheets was investigated under various rolling temperatures and thickness reductions. Results indicated that reasonable matching of rolling process parameters was conducive to realizing the grain refinement and improving microstructure homogeneity. According to the grain size and microstructure homogeneity, the critical rolling process parameters were 400 °C—30%, 300 °C—30%, and 250 °C—40% for AZ31 magnesium alloy sheets. In terms of the basal texture variation, with the increase of thickness reduction, the competition between twinning and DRX resulted in an obvious difference in maximum texture intensity. The former produced the largest maximum texture intensity in 400 °C—20% sheets. However, due to the occurrence of DRX, the maximum texture intensity decreased sharply, but would steadily increase with the growth of DRXed grain. In addition, for DRXed grains, <11-20>//RD texture component would gradually become the dominant texture state, owing to their growth. However, when the deformation continued, <10-10> would rotate toward the RD again. From the fracture elongation perspective, the critical thickness reductions of AZ31 magnesium alloy sheet were 30–40% when the rolling temperature was 400 °C.


Prof. Dr. Ayhan Okçuoğlu Çavdar passed away on June 24, 2019 at Ankara. She served as a researcher and mentor in the Department of Pediatric Hematology and Oncology at Ankara University School of Medicine for over 40 years. After her education at Erenköy Kız Lisesi, Prof. Çavdar graduated from Ankara University School of Medicine in 1953 and completed pediatric medicine residency in 1958. She received a pediatric hematology and oncology fellowship at Washington University. On returning to Turkey in 1961, she established pediatric oncology and hematology units at Ankara University for the first time in Turkey. She made several contributions to oncology in the field of leukemia, particularly orbital granulocytic sarcoma, Hodgkin’s disease, and Burkitt’s lymphoma in Turkish children. Additionally, she researched about pica syndrome in Turkey, thalassemia, hemoglobinopathies, and zinc deficiency in several conditions, especially in pregnancies and new born children with congenital abnormalities.UNESCO Satellite Trace Elements Center in 1998. Prof. Çavdar received many awards from professional societies, including the TUBİTAK Science Award (1976), Pediatric Oncology Service Award (1984), Prof. Dr. Nusret Fişek Public Health Service Promotion Award (1998), and the International Network for Cancer Treatment and Research Award (2007). She published over 475 articles (approximately 197 in Turkish) and authored three books.Dr. Çavdar wrote several research papers on these subjects that were published in international and national periodicals and books. She was elected as the first Turkish member of the Société International d’Oncologie Pediatrique - International Society of Pediatric Oncology. After receiving the Pediatric Board Certificate in 1962, Çavdar became the first Turkish member of the American Pediatric Academy. She was named as the first Turkish pediatric hematologist in the book “Hematology, the Blossoming of a Science,” written by Prof. Dr. Maxwell Myer Wintrobe in 1985. She established three research units, namely, the “Pediatric Hematology and Oncology Research Center,” “Zinc Deficiency Unit,” and “Pediatric Leukemias and Lymphomas Unit,” supported by the Scientific and Technological Research Council of Turkey (TUBITAK). She also constituted Dr. Çavdar defended the idea that the primary role of universities was to educate young generations and to conduct research. On attending international congresses, she shared the scientific knowledge with her colleagues. She was a prominent Figure in the department, rendering full support to us, over the years, in good and bad times. It was an honor and privilege for me to work with her.Çavdar will always be remembered for establishing Pediatric Oncology in Turkey (1961) and as a member of The Turkish Society of Hematology (1967), Mediterranean Blood Club (1975), and The Turkish Academy of Sciences (1993). Her contributions will be greatly appreciated and will evolve where she left.


Japan Primary Care Association has launched an international session at annual conference since 2012. Longitudinal characteristics of the presentations were investigated.Abstracts of the session were analyzed regarding type of presentation , category , and presenters' geographical location.The numbers of oral and poster presentations significantly increased. The most frequent presentation was research with cross‐sectional survey. Majority of presentations were clinical and less educational research, most of which were by Japanese.The numbers of presentations in the international session increased, but the session should be used more rigorously to enhance primary care. The CIA considered active international interaction critical to the responsible mission of the JPCA and determined that presentation of clinical and research work in English would provide opportunities for researchers to enhance their skills in domestic and international healthcare environments. Session organizers and chairs use the CIA sessions as opportunities for meaningful interactions among primary care physicians and trainees in Japan and other countries.Trends in the presentations and the characteristics of the participating presenters are important factors to the CIA in its yearly preparation for the annual conference. Thus, the history of the international sessions provides useful insights for the CIA. The CIA encourages active participation in the sessions by domestic and international primary care scholars, and it believes that it is important to share the session goals and the accomplishments derived from the international session to support the participating scholars. This activity is believed to help them use the international session as a future resource.2The data used for the analysis were derived from the abstracts of the presentations given at the international session of the JPCA annual conference from 2012 through 2018. The conference organizer provided the abstracts. Every abstract was reviewed by the primary investigator (N.H.), and the outcome of the review was confirmed by another investigator (N.B.). Following characteristics were identified: (a) type of presentation , (b) category of presentation , and (c) presenters' geographic area. The research design was also categorized. A descriptive analysis on the type of presentation, category of presentation, and geographic area was performed. Using regression analysis, the study assessed trend on: (a) the total number of presentations across time, (b) oral or poster type, and (c) presenters' national affiliation.3β1 = 4.71, p < .05, R2 = .71). There is also an increase in the number of oral presentations by year .There were eight international sessions at the JPCA annual conference. Initially, they were limited to oral presentations and the poster presentation format was not added until 2015. Altogether, 105 presentations were given: 57 were oral Table  and 48 w3.1Research was the most frequently presented in the session, followed by case report, activity report, and review Figure . CategorMajority of the research were clinical (71%), and the remainder was educational (21%), and the rest included cost‐effective analysis, evaluation of professional behaviors and evaluation of practice. The most frequent type of research design was cross‐sectional (69%), followed by experimental and cohort design . Majority of the educational research employed questionnaire surveys of medical trainees or participants (75%). Regarding the analysis, 55% was quantitative, 29% was qualitative, and the remaining 16% included mixed methods and those with no analysis (methodology presentation).3.2Table 4This study found that the numbers of presentations at the international sessions increased every year Figure , suggestBased on the results, it is timely to consider higher quality research activities for presentation in the international session. Methodological robustness is important to research quality, but the designs used for primary care research might not conform to the established approaches, such as randomized controlled trials and prospective cohort studies. Our results found that more than two‐thirds (69%) of the research used cross‐sectional questionnaire. These designs clearly can be improved to produce higher quality evidence. Further, more methodological variety is needed to answer many of the clinical questions in the primary care field because of the breadth of primary health care. A previous study found that an international research network contributed to research capability.Although the numbers of presenters from around the world with Japanese majority increased, Japanese presentations increased more than foreign ones Table . We shou5We are encouraged by the significant annual increases of international presentations in the international session of the JPCA annual conferences, but several concerns emerged from this study that need to be solved. The international session should be used more for scholarly networking, as methodological learning opportunities, and to present educational research. These weaknesses might be addressed by enriching future sessions, which also might enhance primary care.The authors have stated explicitly that there are no conflicts of interest in connection with this article. Click here for additional data file. Click here for additional data file. Click here for additional data file.