Three-dimensional photonic body surface scanners (3DPS) feature a tool to estimate total body volume (BV) from 3D images of the human body, from which the relative body fat mass (%BF) can be calculated. However, information on validity and reliability of these measurements for application in epidemiological studies is limited.® device using equations by Siri and Brozek. Three scans were obtained per participant . Validity was evaluated based on the agreement of 3DPS with ADP using Bland Altman plots, correlation analysis and Wilcoxon signed ranks test for paired samples. Reliability was investigated in a separate sample of 18 participants  aged 25–66 years using intraclass correlation coefficients (ICC) based on two repeated 3DPS measurements four weeks apart.Validity was assessed among 32 participants  aged 20–58 years. BV and %BF were assessed using a 3DPS (VitusSmart XXL) and air displacement plethysmography (ADP) with a BOD PODMean BV and %BF were higher using 3DPS compared to ADP, , yet the disagreement was not associated with gender, age or body mass index (BMI). Reliability was excellent for 3DPS BV  and good for 3DPS %BF . Results were similar for the standard scan and the relaxed scan but somewhat weaker for the exhaled scan.Although BV and %BF are higher than ADP measurements, our data indicate good validity and reliability for an application of 3DPS in epidemiological studies. Obesity is a major risk factor for non-communicable diseases like type-2-diabetes mellitus and cardiovascular diseases, and accounts for a substantial proportion of disability-adjusted life years and mortality worldwide .The definition of obesity is based on the body mass index (BMI) , 3; howeThree-dimensional photonic body surface scanners (3DPS) provide an opportunity for a standardized acquirement of data on total body volume (BV). This is promising particularly for large-scale epidemiological studies, where standardized, fast, accurate, and precise assessment methods are key issues for a valid estimation of disease risk. In contrast to the abovementioned methods to assess the body composition, the 3DPS examination does not require the participants to be in a fasting state or to restrict physical activity beforehand. In Germany, measurement of body size has been recently conducted for the clothing industry in more than 13,000 individuals using a laser-based 3DPS  . Within The aim of this study was therefore to assess the validity of 3DPS total BV and %BF measures against air displacement plethysmography (ADP) measures and to identify factors potentially contributing to a measurement disagreement in the general adult population. Further, as we have already shown a good short-term reliability of the 3DPS , we examParticipants were recruited from February 2016 to June 2016 as part of the larger-scaled MetSScan study, which targeted a sample size of 500 participants and was conducted by the Molecular Epidemiology Research Group of the Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin. The MetSScan study aims to develop algorithms for the assessment of parameters of the metabolic syndrome [Participants for the main study were recruited based on institution's internal mailing lists, local press releases, newspaper articles, and advertisements as well as public postings. Inclusion criteria were age 18–79 years, sufficient German language skills and the ability to give informed consent. Exclusion criteria were current pregnancy or breastfeeding, known hemophilia, anticoagulant medication, dependency on medical appliances for stand and movement, or current wearing of a plaster cast.For assessment of validity of total BV measurements, ADP measurements were performed on 32 participants in addition to the main study protocol. ADP measurements were not taken in persons with claustrophobia.To assess longer-term reliability  of BV and %BF, the 3DPS measurements were repeated after approximately four weeks in an independent sample of 18 participants under identical conditions.The study was approved by the Ethics Committee of the Charité—Universitätsmedizin Berlin and by the local data protection officer. Informed written consent was obtained from all participants.All participants completed a study protocol at the study center comprising measurement of height and weight, the 3DPS and ADP examination. Due to the original aim of the MetSScan study, we needed a fasting blood sample to determine blood glucose. Thus participants were in a fasting state for at least 8 hours prior to the investigation. All examinations were taken on the same day with a maximum interval of one hour between the 3DPS and ADP measurements.For all examinations, participants wore minimal, tight-fitting, unpadded underwear, no jewelry or eyeglasses and a bathing cap. The measurement of body height followed standardized procedures according to WHO guidelines  using a 2 within a scan time of 10 seconds. Based on the 3D point cloud, 153 anthropometric measures are computed by the software according to international standard DIN EN ISO 20685 [The 3DPS examination was performed with a VitusSmart XXL and the software Anthroscan Professional version 3.3.0 . The measurement principle is based on optical triangulation, whereby the scanner uses eight eye-safe laser sensor heads and cameras to create a 3D point cloud of the human body surface. The measuring range covers 2100 mm height x 1000 mm width x 1200 mm depth. The accuracy is within a level of ±1 mm and the scanner reaches a density of 27 points/cmSO 20685 . FurtherThe study protocol for the 3DPS measurement consisted of three consecutive scans in three different postures: 1. standard position: upright posture with legs shoulder-wide apart and, if possible, no contact of thighs; arms slightly bend and away from the body; hands making a fist with thumbs showing forward; and head positioned compliant with the Frankfort horizontal plane (StdScan), 2. relaxed position: legs hip-wide apart, arms hanging relaxed without contact to the body, hands making a fist, head aligned in Frankfort horizontal plane (RlxScan), and 3. relaxed position after maximum expiration (ExhScan). Except for the ExhScan, participants were asked to breathe normally during the scan procedure. All scan images were immediately visually checked by the study personnel. A scan was repeated if the scan quality did not reach pre-defined standards, e.g. with regard to image artifacts, deficient resolution, or if the posture was not correct. The scan measurement series was cancelled in case the quality remained insufficient after three repeated attempts in the same individual. The 3DPS was calibrated prior to the first examination of each day with a cylinder tube of defined height and width provided by the manufacturer.® , which shows high short-term reliability [® was calibrated each day with a 50 l calibration cylinder, allowing for a maximum standard deviation (SD) of 50 ml in a series of five volume measurements. Body weight was measured with an integrated electronic scale  connected to the BOD POD®, which was calibrated every 14 days with two pre-specified 10 kg weights. Body height was taken from the above described stadiometer measurement. Directly before each measurement, an obligate 2-step calibration was carried out: first, the volume of the chamber was measured without participant inside and then again with the 50 l calibration cylinder placed inside. After the participant entered the chamber, two measurements of raw BV were executed. If these two measurements differed by 150 ml or 0.2% of BV, a third measurement was performed and the two closest measurements within these agreement criteria were averaged. The estimate of raw BV was adjusted for thoracic gas volume (TGV) predicted from gender, age, and height by the software (8) and a body surface area artifact, which arises due to the apparent negative volume of air under isothermal conditions related to the skin surface [corr), body density was calculated as:2) [2) [The assessment of BV by ADP was performed with a BOD PODiability –22, and  surface . Using t0 kg/m2)  or with /m2) [2) :Siri:%B® software.All calculations were automatically executed by the BOD PODThe 3DPS derived BV was corrected for TGV for the StdScan and RlxScan, and residual volume (VR) for the ExhScan, respectively. TGV was calculated according to the following equations by Brozek et al.  and CrapVR was calculated with equations developed by Crapo et al. :Men:VR=%BF was calculated for all corrected BVs of 3DPS scans using Eqs  and (3).2) and was used to define non-overweight , overweight (BMI≥25 to <30 kg/m2) and obesity (BMI≥30 kg/m2) [Data are presented as mean ± standard deviation (SD). BMI was calculated as body weight in kilogram (kg) divided by body height in meter squared (m0 kg/m2) . The com0 kg/m2) , 28. Q-Q0 kg/m2) . Since BMultiple regression analyses were used to predict ADP BV based on 3DPS BV including gender, age, BMI, and an interaction term of 3DPS BV*gender. The final models retained only the significant factors after a stepwise exclusion procedure.For the assessment of reliability, the differences in the duplicated 3DPS measurements (BV and %BF), calculated as scan at date 1 –scan at date 2, and the intraclass correlation coefficients (ICC) ± 95%CI were calculated.P-values presented are 2-tailed and p<0.05 were considered statistically significant. The analyses were performed using SPSS 18  and SAS Enterprise Guide 4.3. .No technical issues or refusal of participants occurred during the completion of the study and quality checks revealed formal plausibility of the data obtained. Basic characteristics of the two subsamples including 32 and 18 participants are summarized in For the comparison of %BF, one participant was excluded from the analysis due to invalidly low %BF (<1% BF) given by ADP. %BF based on 3DPS StdScan and on ADP were highly correlated  as reference methods. A study from Wells et al. with 22 adults using a Hamamatsu Bodyline Scanner reported an underestimation of 0.3 ± 2.7 L against ADP and overestimation of 0.5 ± 2.4 L for UWW, respectively . SimilarTaken together, the calculation rather than measurement of TGV  could have resulted in discrepancies from actual TGV values on an individual level, for which we could not account for. Though this similarly affected both ADP and 3DPS, the ongoing approaches to adjust for TGV differed between methods, e.g. ADP BV was adjusted for 40% of TGV, whereas it was completely deducted from 3DPS BV.The 3DPS does not follow the standardized WHO measurement protocol for assessment of height. In this regard, we previously showed that the VitusSmart XXL 3DPS overestimates body height by on average 0.9 cm compared to manual measurements on basis of standardized WHO guidelines . HoweverFurthermore, other factors independent of the scanner’s ability to validly estimate BV could have contributed to the overestimation of BV. 3DPS visually scans the body surface and thus, any body-enlarging object, like clothing and hair, amplifies BV. Even though this influence was minimized by wearing tight-fitting underwear and a bathing cap, it could not be completely eliminated, e.g. if the participant had a large hair volume, a beard, thick body hair, or padded underwear. In addition, participants may have had the tendency to hold their breath or breathe flatly during the scanning procedure in order to not blur the picture, which could have led to a higher air volume in their lungs.While we observed an acceptable difference between 3DPS and ADP for BV, the agreement between methods found for %BF was lower. A non-zero intercept and non-identity slope in the Bland Altman plots reflected the estimation of %BF on the basis of 3DPS BV to yield higher values than estimation based on ADP. Similar graphical findings were also reported in a study by Wang et al. using underwater weighing as comparison method to evaluate validity of 3DPS based BV and %BF . Bearing2 (as well as for persons with BMI >30.0 kg/m2) Brozek’s [Furthermore, in contrast to BV, for %BF we found a correlation between the difference and the mean of 3DPS and ADP measurements, with higher estimates based on 3DPS than for ADP observed for lower %BF, while with increasing %BF the opposite was true. A similar dependency of the difference between methods and the mean %BF was also found by Wang et al. . This skBrozek’s  instead Brozek’s ; using tIn respect to the reliability of the 3DPS, it should be mentioned that a single assessment of a biomarker, as has often been done in baseline examinations of large-scale epidemiological studies, may be susceptible to short-term variation and not reflect true long-term exposure. Therefore, random measurement error generally tends to decrease correlation and regression coefficients in epidemiological studies toward 0 and bias relative risks toward 1. However, in our analysis, the 3DPS scanner revealed an excellent reliability after a time period of four weeks. To our knowledge, reliability over a longer-time period has not been analyzed previously; however, other studies with consecutive measurements showed very good short-term reliability , 30–32. Our study has some strengths and limitations. ADP has been shown to be an accurate method for assessment of BV, and it is probably the method that is most often used in experimental studies –39. HoweIn conclusion, our study showed good agreement between 3DPS and ADP for measurement of BV and %BF, although measurements were slightly higher for 3DPS compared to ADP. Further, we found high ICCs for BV and %BF when measured with 3DPS four weeks apart. These data indicate good validity and excellent four-week reliability for an application of 3DPS to assess BV and %BF in an epidemiological study.


First of all, let me wish you a healthy and wonderful year in 2018.When we look back on the past year, all of us are proud to announce that 2017 has been a highly fruitful year for our journal. This is wonderful news, rewarding the continuous support offered and confidence expressed by all our authors, readers, reviewers, members of the editorial board, as well as the efforts of our editorial team. Pharmaceuticals obtained a score of 4.90 for 2017, which propels it to #8/168 in the category Pharmaceutical Science and #21/158 in the category Molecular Medicine.The most noteworthy fact is, undoubtedly, the release of the CiteScore™ values  by ElsevImportantly, also, our journal is now indexed in the Emerging Sources Citation Index .Pharmaceuticals. Incidentally, have you noticed that each of the four issues of Volume 10 (2017) is now illustrated by an elegant colorful cover? [Consequently, it is not surprising that the abstracts of publications in 2017 have been seen more than 400,000 times [l cover? .Atanas G. Atanasov ;Connor R. Caffrey ;Antoni Camins Espuny ;Michael Danilenko ;Christophe Dardonville ;Gunars Duburs ;Jesus Jimenez-Barbero ;Carlos Alberto Manssour Fraga ;Susan Matthews ;Mary J. Meegan ;Pascal Sonnet ;Arpad Szallasi ;Peter Timmerman .During the past year, twelve distinguished scientists accepted to act as editorial board members, namely:  have joined Annie Mayence  as Associate Editors.Joachim Jose  and Klaus KopkaEmerging theranostic tracers in the context of radiopharmaceutical drug development , K. KopkNutraceuticals and botanicals: bioactive molecules and therapeutic properties for human health , D. DonnSelected papers from the 2nd International Electronic Conference on Medicinal Chemistry , J.J. VaGlycosaminoglycans and proteoglycans , B. MullTissue protective agents: new drugs and technologies , S. CollEpilepsy and neurodegeneration: current therapeutic implications , A. CamiIn 2017, we completed six remarkable Special Issues on the following subjects:An Updated View on an Emerging Target: Selected Papers from the 8th International Conference on Protein Kinase CK2” [Identification and Characterization of Antimicrobial Peptides with Therapeutic Potential” [Two books have been released: “ase CK2”  . hemistry . The Conhemistry . MentionThe topical section “Choices of the Journal” , createdMaintaining the high quality of our publications and increasing the visibility of our journal remain essential targets on our road map. We are already working in that direction and we are collecting applications for a Travel Award for a PhD student to attend a conference in 2018 . Moreove


Esterification is one of the most important methods to alter the structure of starch granules and improve its applications. Conventionally, starch esters are prepared by conventional or dual modification techniques, which have the disadvantages of being expensive, have regent overdoses, and are time-consuming. In addition, the degree of substitution (DS) is often considered as the primary factor in view of its contribution to estimate substituted groups of starch esters. In order to improve the detection accuracy and production efficiency, different detection techniques, including titration, nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis/infrared spectroscopy (TGA/IR) and headspace gas chromatography (HS-GC), have been developed for DS. This paper gives a comprehensive overview on the recent advances in DS analysis and starch esterification techniques. Additionally, the advantages, limitations, some perspectives on future trends of these techniques and the applications of their derivatives in the food industry are also presented. Starch is an inexpensive, abundantly available, widely used, and naturally produced polysaccharide molecule found in fruits, seeds, stems, tubers and roots for the storage of solar energy . It exisThese dual modification techniques, based on conventional methods, have been optimized by the use of different cultivars , pH 11]11], fattThe reaction mechanism of esterification is shown in There have been few publications focusing on the manufacture and application of starch esters within the food industry. Both Zukowska et al.  and AbbaThis review, attempts to redress the balance, summarizes recent advances in esterification procedures, including conventional and dual modification, such as catalyst addition, ball milling activation, high temperature/pressure, microwave assist, pulsed electric fields, chemical synthesis involving cross-linking, and oxidation.A commonly used parameter when considering the quality and performance of starch esters is the DS, which relates to the average number of hydroxyl groups replaced by acetyl groups or other derivatives per glucose unit and serves as an indicator of the its properties and applications ,38. The 2 that is present in the atmosphere, as it can dissolve into the solution causing non-avoidable experimental error. Back-titration, however, relies on the saponification of the product using NaOH or KOH to hydrolyze the acetyl groups followed by titration of the excess alkali which thus avoids disturbing of CO2 in the atmosphere therefore removing this source of error [Titration and back-titration based on color change of the indicator are both widely accepted and used in industrial, as well as laboratory, settings ,39,40. Mof error , however1H-NMR and 13C-NMR may be employed for evaluating succinate, alkenylsuccinate and some acetate esters by adding sodium 3-(trimethylsilyl) propionate 2,2,3,3-D4 as an internal standard [−1) related to A2929 of starch CH2 vibration (2929 cm−1) were regressed to predicted other samples. 1H-NMR and FT-IR analyses require complicated procedures however, more and more researchers employ this method to determine the precise DS value [NMR and FT-IR are systematic methods that have been used to measure the DS of starch esters. standard ,41. FT-Istandard . ResearcDS value ,43,44. TDS value .2, which was produced by neutralization of carboxyl groups and the added bicarbonate solution in sealed vials. Therefore, HS-GC is another simple and automated method for determining DS of anhydride-modified cellulose, which can be used in assessing the DS of starch esters.Another method that has been employed to estimate DS of anhydride modified cellulose is the combination of headspace and gas chromatography (HS-GC) proposed by Zhong et al. . The resStarch esters modified by conventional method have already been widely applied in industry by converting the hydroxyl groups of starch into diverse derivatives. There is a mass of factors affecting the end products with different DS, such as starch cultivar, reactant concentration , acid or alkali, reaction time, temperature, pH moderation, reaction medium and the presence or absence of a catalyst, which is illustrated in Different starch cultivars exhibit different amylase/amylopectin ratios. As early as 1996, Fringant et al.  studied In recent years, in order to develop more applications of esterified starch, researchers have focused on health care applications, as well as new composite materials for nanocomposite film development ,56. FoodResearch has used these starch esters with lower DS of 0.01–0.20 in applications for food formulation and other areas, such as film forming, binding, adhesion, thickening, stabilizing and texturing ,50,59. SDual modification is the combination of conventional modification and other kinds of techniques to improve the properties of starch esters. These dual modification methods include chemical reaction in the presence of a specific physical environment or an enzymatic treatment to increase the yield of derivatizations or improve the DS . The comA catalyst is an active factor that accelerates reaction rates and shortens reaction times. According to the origin of the catalyst, it can be divided into two types involving chemical reagents or enzymatic materials.p-toluenesulfonic acid as a catalyst to produce starch esters had a great effect on the DS value, which played an essential role on increasing the collision times of molecules among starch and acetic anhydride, resulting in higher DS.Acetylated starch with a low DS of 0.01–0.20 is usually studied in an aqueous medium in the presence of alkaline catalyst. In recent years, the introduction of a catalyst-used reaction was proposed to increase DS by Wang et al. . Sodium Use of an enzyme as a catalyst is an alternative way to accelerate reaction rates and improve reaction efficiencies. Singh et al.  considerTo sum up, even though the use of an organic solvent as catalyst can improve the reaction efficiency, the challenges of solvents’ poor environmental image and high cost make it difficult to expand extensive application into industry. By contrast, the enzyme catalyst-assisted methods are environmentally friendly with few by-products and can therefore be used directly.Ball milling, as a lower-cost and physically-modified method, has been used to enlarge the surface area of molecules. Friction, collision, impingement, shear or other mechanical actions are induced during ball milling , these mOther effective, physical methods to active starch granules utilize harsh circumstances, such as high temperature or high pressure. Shogren  discoverMicrowaves are known to be non-ionizing energy, which can generate heat in the penetrated medium by the “molecular friction” in an alternating electromagnetic field. When microwave technology was used in esterification, Jyothi et al.  combinedMore researchers and entrepreneurs are choosing to employ ecologically friendly procedures that use less volatile organic solvents, avoid toxic solvent consumption, and provide better economic benefits. In view of the above research, due to its energy saving, high conversion rate, speed of reaction and existing applications, microwave-assisted esterification has many advantages that make it a popular, emerging and promising technique.Pulsed electric fields (PEF), as a promising non-thermal technique, has been widely used for inactivation of microorganisms and enzymes in food applications. In addition to this, however, researchers have reported the effects of PEF treatment on starch modification with different types, including A-type (corn starch), B-type (potato starch) and C-type (cassava starch) ,4. TheseThus, PEF, combined with esterification, could be a novel and promising technique to improve reaction conditions and efficiency by modifying its properties. There are still many challenges for PEF–assisted esterification, including understanding the transformation of the fractal nanostructure, however, there are also opportunities for more applications in new areas.The combination of esterification with other chemical modification methods, such as cross-linking and oxidation, have already been studied. These starch esters are prepared by synthetic methods and, due to double functional groups being present in the starch molecules, dual properties are exhibited.Simple modification of cross-linking, where the polymer chains of starch cross-link with two- or poly-function compounds, such as adipic acid, citric acid or phosphoric acid, leading to a significant increase in molecular weight, is a well established and widely used chemical method for starch modification in the laboratory, as well as in industry .The integration of esterification and cross-linking is an effective way to obtain the derivatives with double functions. Han et al.  exploredBased on the mentioned studies, therefore, it appears that it is still necessary to take up the challenge to establish novel methods in order to produce starch esters with double functional groups with a high rate of efficiency, so as to cater to a more universal applications in different areas.Oxidation as a form of chemical modification, involves the introduction of carboxyl and carbonyl functional groups by means of sequential depolymerization of the starch. Such starches have been established to be whiter in color, but have low values in breakdown and setback viscosity when pasting properties are analyzed . There aThe preparation of starch esters is a well-established field of research. The microwave-assisted method mentioned above is very convenient and eco-friendly. As previously noted, conventional esterification of starch is usually done under high pH, using harsh chemicals and creating toxic waste, making it expensive and potentially unavailable for large-scale industrial production of modified starches. Unlike conventional modification, dual modification methods, such as PEF–assisted, microwave assisted or enzyme catalyzed esterification, are multifunctional, manageable, less time consuming and have a high reaction efficiency. In addition, these methods avoid the use of organic reagents, which are mild to the environment, as well as the end product. Enzyme catalyzed esterification, however, is much more expensive than the other two methods. Therefore, the dual methods of PEF–assisted or microwave-assisted esterification are potential and promising techniques for starch modification. Nevertheless, the combination of the alternative promising synthesis procedures and its nanostructure, characteristics and novel applications are still far from being exhaustively investigated.This review summarizes current determination methods of DS, the merits and shortcomings of diverse modification methods, and the advanced applications for starch esters. From these methods, these emerging dual techniques appear to have the capability to improve DS value, accelerate reaction efficiency, shorten reaction time and explore novel applications. In addition, the dual modification techniques overcome the drawbacks that occur in conventional modification, and can produce all the advantages when modified by the single method, which would gain considerable economic benefits. The conclusion is that dual modification techniques of microwave-assisted and PEF–assisted esterification, which exhibit cost reduction, reagent saving, reaction time shortening, and reaction efficiency promotion, are the two main promising trends for starch modification.


Lactobacillus was dominant flocculating microorganism in sour liquid. In total, 86 bacteria, 20 yeasts, and 10 molds were isolated from the sour liquid and only eight Lactobacillus species exhibited flocculating activity. Lactobacillus paracasei subsp. paracasei L1 strain with a high flocculating activity was isolated and identified, and the mechanism of starch flocculation was examined. L. paracasei subsp. paracasei L1 cells formed chain-like structures on starch granules. Consequently, these cells connected the starch granules to one another, leading to formation of large flocs. The results of various treatments of L1 cells indicated that bacterial surface proteins play a role in flocculation and L1 cells adhered to the surface of starch granules via specific surface proteins. These surface starch-binding proteins were extracted using the guanidine hydrochloride method; 10 proteins were identified by mass spectrometry: three of these proteins were glycolytic enzymes; two were identified as the translation elongation factor Tu; one was a cell wall hydrolase; one was a surface antigen; one was lyzozyme M1; one was a glycoside hydrolase; and one was an uncharacterized proteins. This study will paves the way for future industrial application of the L1 isolate in starch processing and food manufacturing.In the current study, we focused on the mechanism underlying starch flocculation by the sweet potato sour liquid. The traditional microbial techniques and 16S rDNA sequencing revealed that  The processing of starch flocculated with sour liquid has a 400-year history in China . Sour liStreptococcus lactis from a mung bean sour liquid was isolated, and shown to bind to the starch granules, and promote the flocculation of starch . The sour liquid (1 L) was collected in a sterile culture flask and transferred to a laboratory at 4°C. Microorganisms were plated for enumeration and isolation on the same day. Sweet potato starch was purchased from Shandong Bio Sunkeen, Co., Ltd. . Phosphate-buffered saline  was purchased from Sigma Chemical, Co. . All other chemical reagents were of analytical grade.The sweet potato juice medium was prepared as follows. Sweet potato infusion was prepared by boiling 200 g of sliced (washed but unpeeled) sweet potatoes in 1 L of distilled water for 30 min, and decanting or straining the broth through cheesecloth. Distilled water was added such that the total volume of the suspension was 1 L; 20 g of glucose, 2 g of lactose, 5 g of yeast extract, and 5 g of sodium acetate was then added, and the medium was sterilized by autoclaving at 115°C for 15 min.vide infra), cultured at 30°C , and were then stored at -4°C to screen the strains with high flocculating activities  media supplemented with cycloheximide (50 μg/mL) to inhibit fungal growth . The plativities .The selected strain slopes were inoculated into 5 mL of sweet potato juice medium and cultured at 30°C. Bacteria and yeasts were cultured for 1 and 3 days, respectively. The molds were cultured with shaking at 160 rpm for 5 days. Then, the flocculation rate of fermentation liquor was determined as the flowing methods. Using the flocculation rate of the cultures as an index, with the sweet potato juice medium as a control, the strains were screened for high flocculating starch activity.Distilled water (100 mL), 0.5 g of sweet potato starch, and 5 mL of the liquor to be tested were placed in a 150-mL beaker. The liquid was agitated for 3 min on a magnetic stirring apparatus, and then left to stand for 3 min. As a control, sweet potato juice was used instead of the fermented liquor. The flocculation efficiency was expressed as FR, by measuring the decrease of turbidity of the upper phase . FR was Where A and B are optical densities of the control and sample, respectively, at 550 nm.Microbial genomic DNA was extracted from 1 mL of the sour liquid by using the TIANGEN DNA stool mini kit  according to the manufacturer’s instructions. The V4 variable region of 16S rDNA was amplified using the universal primers 520F (5′-AYTGGGYDTAAAGNG-3′) and 802R (5′-TACNVGGGTATCTAATCC-3′) . The PCR1, which required that the overlap of reads 1 and 2 was ≥10 bp, without any mismatches. The reads that could not be assembled were discarded. Chimera sequences were removed using UCHIME in mothur (version 1.31.22).Sequence reads were excluded from analysis if their length was less than 150 bp, the average Phred score was lower than 20, contained ambiguous bases, a homopolymer run exceeding six bases, or when mismatches in primers were detected. Sequences that passed quality filtering were then assembled by Flash3; the sequences were clustered into operational taxonomic units (OTUs). The longest sequence in each cluster was selected as the representative. The taxonomy of each OTU was assigned by BLAST-searching the representative sequence against Greengenes reference database (Release 13.84)  .g. Each cell pellet was resuspended in 0.5 mL of dH2O, and DNA was extracted using the TIANGEN DNA stool mini kit  according to the producer’s instructions.Pure isolates were grown to a late stationary phase in 5 mL of media. The cultures were centrifuged for 10 min at 4,000 × Full-length 16S rDNA amplicons were generated with bacterial primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-CTACGGCTACCTTGTTACGA-3′). The PCR amplification and sequencing were performed, as described previously .5.To identify 16S rDNA sequences most similar to the obtained sequences, all sequences were matched against nucleotide sequences deposited in GenBank using the BLASTn programFinally, strain identification based on its colony character, morphological, and physiological characteristics, as well as 16S rDNA sequence homology referencing Bergey’s Manual of Systematic Bacteriology.Lactobacillus paracasei subsp. paracasei L1 slope were inoculated into a tube of fermented sweet potato juice (5 mL) and were cultured for 24 h at 30°C. Then, the inoculum was inoculated into the sweet potato juice medium  and cultured for 24 h at 30°C.g for 10 min. The cell pellet was washed twice with distilled water; 5 mL of distilled water was added to obtain a bacterial suspension. Distilled water (100 mL), 0.5 g of sweet potato starch, and 5 mL of the bacterial suspension were placed in a 150-mL beaker to test the flocculation activity.Flocculation activity of L1 in fermentation liquor was as follows. Five milliliter of the fermented liquor were centrifuged at 4,000 × Lactobacillus paracasei subsp. paracasei L1 culture (10 mL) was centrifuged at 4,000 × g for 10 min. The cell pellet was washed twice with dH2O, and 10 mL of distilled water was added to obtain a bacterial suspension. Flocculating activities of culture broth, cell-free supernatant and cell pellet were tested (e tested .Lactobacillus paracasei subsp. paracasei L1 cultures (10 mL) were centrifuged at 4,000 × g for 10 min. The cell pellet was washed twice with distilled water; 10 mL of distilled water was added to obtain a bacterial suspension. Distilled water (100 mL), 0.5 g of sweet potato starch, and 5 mL of the bacterial suspension were placed in a 150-mL beaker. Microtrac laser particle size analyzer  and laser diffraction particle size distribution meter were used to determine the particle size distribution of sweet potato starch before flocculation. Thereafter, the liquid was agitated for 3 min on a magnetic stirring apparatus, and then left to stand for 3 min. Microtrac S3500 was next used to determine the particle size distribution of starch and in the supernatant after flocculation  in 0.01 M phosphate buffer, pH 7.2] on brass stubs and chromium-coated by Xenosput 2000 chromium coater with the deposition parameters of 0.06 sputter Amps for 40 s. Coated preparations were visualized with Hitachi S4800 scanning electron microscope  at the accelerating voltage of 2 kV  was weighed and tested by zeta potentiometer . The values of ζ potential of sweet potato starch milk, termined .Lactobacillus paracasei subsp. paracasei L1 cells were cultured for 24 h in the sweet potato juice medium, washed twice with PBS, and collected by centrifugation. The specific experiments were performed as follows: cells were resuspended in PBS to 8 log CFU/mL and then heat-treated in a water bath at 30, 40, 50, and 60°C for 30 min. Next, cells were resuspended in PBS to 8 log CFU/mL and then irradiated using an ultraviolet lamp . Cells were resuspended in PBS to 8 log CFU/mL and then placed in an ice-water bath and sonicated for 5 min . Processing was stopped when the temperature reached values ≥ 15°C. When cells cooled to below 10°C, they were again treated for 5 min. When cells subsequently reached a temperature of 10°C, treatment was repeated for 5 min. Cells were pretreated with 3% trichloroacetic acid or 10-4 mol/L lithium chloride at 28°C for 30 min. Cells were resuspended in PBS to 8 log CFU/mL and then separately pretreated with the following enzymes: trypsin  ml-1, at pH 7.5 for 6 h at 37°C; α-amylase  ml-1 at pH 7.0 for 4 h at 40°C; lysozyme  ml-1 at pH 6.0 for 1 h at 37°C. The flocculating experiment was performed using 3 g of Tween 80 L-1. Then, the flocculating experiment was performed using 5 g/L of glucose or maltose , and washed three times with PBS. The cells (1 g) were incubated in 20 mL of 4 M guanidine hydrochloride with shaking (at 200 rpm) for 60 min at 37°C. The supernatant was collected after centrifugation at 12,000 × g and 4°C for 10 min; it was dialyzed overnight in a dialysis bag, with PBS as the dialysis solution. PBS was replaced 5–6 times. Sweet potato starch (0.5 g) were added to 10-mL samples of the supernatant, shaken for 30 min to ensure full exposure of granule surface to the supernatant, and then washed with PBS and centrifuged , three times, to remove unbound proteins. To extract starch-bound proteins, each starch pellet was resuspended and incubated for 5 min in PBS (control) and PBS containing 100 mM maltose. The starch was removed by centrifugation and the supernatant from each tube was separated and purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified samples were digested with trypsin, analyzed by high performance liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) using Q Exactive (Thermo Scientific), and identified by Mascot 2.3.0 using the Uniprot Lactobacillus database6 (atabase6 . The idep < 0.05) among obtained results using the Student’s t-test or ANOVA followed by Duncan’s multiple range test. All data were analyzed using the SPSS 16 software .Data were obtained in triplicate and are reported as averages; Statistical analyses were performed to determine significant differences (-1). Yeasts were 5.04 ± 0.04 log CFU mL-1. Furthermore, molds were 2.71 ± 0.02 log CFU mL-1 and the lowest number in sour liquid. Bacteria, therefore, were dominant in the sour liquid.The counts of bacteria was the highest . We conclude that Acetobacter, Pseudomonas, and Lactobacillus are the dominant bacteria in the sour liquid.To investigate the dominant bacteria in sour liquid, bacterial composition in the sour liquid was evaluated using high-throughput sequencing of the V4 regions of 16S rRNA genes. The bacterial community was analyzed at the genus level by comparing with Greengenes reference database. Lactobacillus by 16S rDNA sequence homology comparisons. The yeasts and molds did not show any starch-flocculating activity, indicating that Lactobacillus was the dominant flocculating microorganism in the sweet potato sour liquid. The most pronounced flocculating activity among the flocculating strains was observed during the fermentation with Lactobacillus strain L1. Strain L1 was subsequently identified as L. paracasei subsp. paracasei based on its colony character, morphological, and physiological characteristics, as well as 16S rDNA sequence homology referencing Bergey’s Manual of Systematic Bacteriology; accordingly, it was named L. paracasei subsp. paracasei L1. This strain was deposited in the China General Microbiological Culture Collection Center .In total, 86 bacteria, 20 yeasts, and 10 molds were isolated from the sour liquid on TJA and Rose-Bengal media; they were inoculated and cultured in the sweet potato juice medium to identify strains with flocculating activity. Eight strains exhibited flocculating activity. These eight isolates were all bacterial strains, with a rod shape and chain-like arrangement of cells, and were identified as L. paracasei subsp. paracasei L1, was investigated. It was conclude that more than 85% of the flocculating activity was cell-associated, and less than 15% of the activity was associated with the extracellular secretions .The distribution of flocculating activity in cell culture, i.e., its association with the cells and extracellular secretions of L. paracasei subsp. paracasei L1 cultures was evaluated with Microtrac S3500 laser grain size analyzer. The average particle size, D50, also called the median diameter, denotes a cumulative 50% point of diameter (or 50% pass particle size). The D50 (the average particle size) of starch granule size increased, from 2.286 to 5.450 μm, in the presence of L. paracasei subsp. paracasei L1 cells, suggesting that the starch granules formed massive floccules .The size of starch granules before and after treatment with Figures 4A,B). In the presence of bacteria, the starch granules rapidly aggregated and formed massive floccules. Furthermore, SEM analysis revealed that the L. paracasei subsp. paracasei L1 cells adhered to the surface of starch granules . Multiple bacterial cells adhered to starch granules surface and also to each other, thus forming bridge-like structures linking starch granules and forming the aggregated floc.The size distribution of starch granules before the addition of bacteria was homogeneous and uniform, as observed under an optical microscope at a magnification of 100× (L. paracasei subsp. paracasei L1 cell suspension in sweet potato juice medium (Table 1). After the addition of cells to the starch suspension , the potential was closer to zero. According to the DLVO theory, this indicated that the starch granules were in a very unstable state and readily formed a floc precipitation. Both the starch and the cells were negatively charged in water, indicating that the starch-bacterium adhesion was not effected by electrostatic interactions.ζ Potential of the starch suspension was initially a lot lower than that of Figure 6, the flocculating activity was sensitive to heat treatment, but not to α-amylase or lysozyme treatments, indicating that the surface polysaccharide of L. paracasei subsp. paracasei L1 did not mediate the flocculation. In contrast, the flocculating activity of cells was affected by UV, ultrasonic treatment, trichloroacetic acid, and lithium chloride, which are all protein denaturants. Furthermore, trypsin treatment significantly reduced the flocculating activity of cells. Collectively, these results indicated that bacterial surface proteins play a role in flocculation.The effect of various treatments on the flocculating ability of L1 cells was investigated to determine the nature of the flocculation factor on the cell surface, and the nature of the interacting force between the cells and starch granules. As shown in L. paracasei subsp. paracasei L1 cells and starch granules; similarly, electrostatic interactions did not occur because both starch granules and cells were negatively charged in water (vide supra). Moreover, these interactions were likely specific because glucose and maltose significantly inhibited the flocculation .Flocculation was slightly affected by Tween 80. This suggested an absence of hydrophobic interactions between L. paracasei subsp. paracasei L1 extracted by the guanidine hydrochloride method were incubated with starch granules. After a series of non-specific washes, proteins adhering to the starch granules were removed by re-suspension in PBS containing 100 mM maltose. Ten candidate starch-binding proteins were then identified by LC-ESI-MS/MS: three of these proteins were glycolytic enzymes; two were identified as the translation elongation factor Tu; one was a cell wall hydrolase; one was a surface antigen; one was lyzozyme M1; one was a glycoside hydrolase; and one was an uncharacterized proteins (Table 2). Mascot score for these hits was >100. None of the proteins were detected in the control.Surface proteins from Acetobacter, Lactobacillus, and Pseudomonas are dominant in the sweet potato sour liquid, yet all the strains screened in the current study that exhibited flocculating activity belonged to the Lactobacillus genus. Notably, Lactobacillus sp. were also the first species discovered to flocculate starch  was detected on the L1 cell surface (Mascot score of 183). These hydrolases play important roles in the regulation of cell wall growth, turnover, and maintenance, and in the separation of daughter cells. Hydrolase is also found on the cell wall of Lactobacillus rhamnosus GG (LGG), often near the mature septa of exponential cells, exhibiting D-glutamyl-L-lysyl endopeptidase activity in zymogram assays , which indicated that this protein was highly likely to be as identified. Cell wall hydrolases catalyze the cleavage of peptidoglycan sugar or peptide chains . Similarm assays . Mutatiom assays . In thisB. thetaiotaomicron, a prominent mutualist in the distal intestine of adult human , enolase , and glyceraldehyde-3-phosphate dehydrogenase ; the fourth one was identified as a glycoside hydrolase (score 139). These sugar-metabolizing enzymes are found in most bacterial cells and play a role in sugar catabolism or degradation of such complex carbohydrates as lactose or starch . We askelt human . Whole-g pathway . SimilarLactobacillus was the dominant flocculating bacterial genus in the sweet potato sour liquid. L. paracasei subsp. paracasei L1 strain with a high flocculating activity was isolated, and the flocculation mechanism of its adhesion to starch was investigated. Our results showed that the L. paracasei subsp. paracasei L1 cells specifically bound starch granules and linked these starch granules to form large flocs by bridging. This accelerated starch deposition. The starch-binding proteins on the surface of L. paracasei subsp. paracasei L1 cells were extracted using guanidine hydrochloride, and 10 proteins with Mascot scores ≥ 100 were identified by mass spectrometry. These proteins are also present, as adhesion molecules, on the cell surface of other probiotic bacteria. Their role in bacterial starch metabolism, functional properties, and potential applications in adhesion to starch or other materials should be further investigated.As determined by 16S rDNA sequencing and traditional microbiology techniques, Lactobacillus by 16S rDNA sequence homology comparisons. Sequences of this project have been deposited in the NCBI sequence read archive (https://www.ncbi.nlm.nih.gov/genbank/) under GenBank accession numbers: KY952217 (L1); KY952218 (LL1); KY978461 (L28); KY978462 (L36); KY978463 (L53); KY978464 (S07); KY978465 (S10); KY978466 (S32).Eight strains exhibited flocculating activity. These eight isolates were all bacterial strains, and were identified as LZ, YX, and XhL performed all experiments and wrote the paper. YY, XoL, HZ, and ZZ conducted the experiments and data analysis. All authors read and approved the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer GP and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


The Republic of Kazakhstan is in transition to the World Trade Organization (WTO) accession. Accessing to WTO has always been and remains a foreign policy priority for Kazakhstan. As a part of this transition, Kazakhstan is identifying new directions for developing and reforming healthcare disciplines. The goal of this overview is to provide information about the work of centers of sanitary and epidemiological expertise to medical and public health professionals in various fields. Additionally, the goal of this paper is to provide information about international accreditation and quality control activities of the centers.There are 16 centers of sanitary and epidemiological expertise in the Republic, 15 of which have been locally accredited. Implementation of innovation policy at the centers for sanitary and epidemiological expertise fueled the process of international accreditation. It is important to point out that the only center which has an international accreditation as of September of 2012 is the Astana city center.The center for sanitary and epidemiological expertise of Mangistau region was accredited in 2010 and gained the status as a test center. The Mangistau test center applies previously developed criteria and procedures to estimate technical competence. One of the main standard documents used to estimate competence is ISO/IEC 17025 from the International Organization for Standardization/International Electrotechnical Commission, which assesses the following factors:- personnel technical competence- validation of testing methods- efficiency, calibrating test and service of testing equipment- testing environment- selection, processing and transportation of testing samples- quality provision of testing and calibration dataThe introduction of a quality management system was a success. In order to increase the technical competence level, 29 specialists from the center have been trained and have received certificates of compliance.In September 2011 and April 2012, the Ministry of Healthcare of the Republic of Kazakhstan within the framework of the World Bank project, component F (“Food Safety Program and World Trade Organization accession”), arranged international workshops aimed to train quality managers to meet international requirements and standards. Training seminars for international accreditation provided hands on and methodological sessions combined with the dissemination of the comprehensive tutorial materials. Regional trip seminars were organized by the center specialists.Laboratories of the Republic were accredited by the National Center for Accreditation (NCA). On October 27, 2010, NCA signed the International Laboratory Accreditation Cooperation Mutual Recognition Arrangement (ILAC MRA) agreement on mutual international acceptance of ILAC standards.All accredited testing centers for sanitary and epidemiological expertise have an agreement with the National Accreditation Centers regarding the application of the Laboratory Combined MRA Mark. It allows for laboratories to apply this quality brand where applicable on internationally accepted test procedures and measurements. Laboratory test records are issued with the NCA logo and ILAC MRA marking. All these procedures are directed to achieve mutual cooperation with international organizations as a part of the World Trade Organization accession. The international acknowledgement of the national accreditation system will serve as the basis for bilateral arrangements between Kazakhstan and its trade partners, which will create favorable conditions for entering WTO.


The purpose of this study was to compare the outcomes of the buccal mucosa and lingual mucosa used in children who received multiple failed hypospadias surgeries.We conducted a retrospective study of 62 children who received buccal or lingual mucosa graft urethroplasty in our hospital between 2012 and 2015. The ages ranged from 3.5–11 y. All cases included multiple failed hypospadias procedures, and the subjects received previous operations 2–3 times. All patients underwent one-stage operations. Thirty-three cases were treated with lingual mucosa grafts. The patient ages ranged from 3.5 to 11 y (median 7.5 y), and they had previous operations 2–3 times (mean 2.8±0.7). Grafts ranged from lengths of 2–6 cm (mean 5.1±0.46 cm) and widths of 0.5–1.5 cm (mean 1.2± 0.16 cm). Our follow-up was 5 to 12 m (mean 8.3±1.2 m). Twenty-nine cases were treated with buccal mucosa grafts. The patient ages ranged from 4 to 9.2 y (median 7.0 y), and they had previous operations 2–3 times (mean 2.5±0.2). Grafts ranged from lengths of 2–5.3 cm (mean 4.9± 0.28 cm) and widths of 0.5–1.5 cm (mean 1.0±0.11 cm). Our follow-up was 5 to 12 m (mean 7.9±0.5 m). The results were tested with SPSS 18.0. The rates of complications were compared by a chi-square test, and pre-operative conditions were compared by t test.For the outcomes of the two groups, there was no significant difference between the groups in terms of age, preoperative surgery time, and the length and width of the grafts (p>0.05). For the lingual mucosa graft group, fistula: 2/33 (6.0%), stricture: 1/33(3.0%), ventral curvature: 2/33(6.0%), complications: 5/33(15.0%), success rate: 28/33(84.8%), Hose score: 14.34±0.95, peak flow: 6.5 ml/s-12.0 ml/s, and mean peak flow: 9.3±0.4 ml/s. For the buccal mucosa graft group, fistula: 2/29(6.8%), stricture: 2/29(6.8%), ventral curvature: 1/29 (3.4%), complication rate: 5/29(17.0%), success rate: 24/29 (83.0%), Hose score: 14.28±1.03, peak flow: 6.5 ml/s-12.0 ml/s, and mean peak flow: 9.2±0.2 ml/s. There were no differences between the two groups for overall success, complication rates, peak flow, and the Hose scores(P>0.05).The lingual mucosal graft and the buccal mucosa graft both achieved good outcomes, and the lingual mucosa graft made up for the shortcomings of the buccal mucosa graft, which provided a reliable way to treat the multiple failed hypospadias surgeries in pre-pubertal boys. The incidence of hypospadias is approximately 1/300, which has rapidly grown in recent years. Because surgeries are the only way to solve hypospadias, many studies of surgeries for the condition have been reported in the literature. We found different types of surgeries with different outcomes . Since HWe recruited sixty-two pre-pubertal boys who received buccal or lingual mucosa graft urethroplasty in our hospital between 2012 and 2015. The patThe preparation of the LMG before the operation was similar to BMG urethroplasty. Because all of the 62 patients could have received either surgery, we assigned the two procedures at different periods. All patients were operated on general anesthesia with nasotracheal intubation. We documented the appearance of the failed hypospadias procedures . For theHarvesting method for the lingual mucosa grafts: We measured the urethral defect , keepingHarvesting method for the buccal mucosa grafts: We measured the defect and fully exposed the lower lip mucosa. Nor-epinephrine  was applied to the area in both groups, and the graft was harvested Figs  and 8.The graft was defatted . The donThe urethral catheter was removed after 3 weeks. For data analysis, our hospital used IBM SPSS version 18.0 for statistical analysis. The comparison of urethral meatus measurements and the outcomes was performed with a chi-square test, and P < 0.05 indicated that the differences were statistically significant. We assessed the outcome of the cosmetic results with the HOSE score table , diverticula (which could affect the urine stream and require surgery), meatal stenosis/stricture (with stricture apparent on a urethrogram and requiring subsequent intervention), persistent ventral curvature, glans dehiscence and/or skin reoperation with some bleeding, and a loss of stent .For the outcomes of the two groups, there was no significant difference between groups in terms of age, previous repairs, and the length and width of the grafts (p>0.05) . For theIn our study, 10 cases were defined as failures based on the definition of success. Three patients were diagnosed with urethra strictures  based on the urethrography. All returned to normal after receiving dilation. Four patients with fistulas had received fistulous neoplasty. Three cases with ventral curvature  were conducted with dorsal tunica albuginea plication based on the methods of Baskin (1994) [Urethroplasty is the key procedure for treating hypospadias and different types of procedures have appeared over the years. Many patients have to face multiple surgeries due to failed procedures, and these patients typically develop problematic conditions, such as stricture, fistula, and ventral curvature , 12. It However, this method is not suitable to patients with longer urethral defects, physiological deficiency of oral grafts or oral mucosa damage due to previous harvestings. Additionally, many scholars have reported that the buccal mucosa graft donor site has some complications, such as persistent perioral numbness, salivary changes, and difficulty in opening the mouth . This diWe could barely find any studies about the comparison between the two grafts used in inlay urethroplasty, especially in the pre-puberty boys from a Chinese population. To evaluate the outcomes, we conducted a retrospective study of sixty-two children who received either buccal (twenty-nine) or lingual (thirty-three) mucosa graft urethroplasty in our hospital between 2012 and 2015. The ages ranged from 3.5-11y. For the outcomes of the two groups, there was no significant difference between groups in terms of age, preoperative surgery time, and the length and width of the grafts (p>0.05) . For theOur results showed that we have achieved satisfactory outcomes for the two groups. The overall successful rates of the BMG and LMG were 83.0% and 85.0%, respectively, which were similar to the 87% success rate reported by Abdelhameed et al  in adultWe know from the literature that the lingual mucosa shares many characteristics with the urethra, such as the same embryologic origin as the urethra as well as the immunologic advantage of resistance to infection and tissue characteristics , which are more obvious prior to puberty , 21. TheWhen multiple hypospadias procedures fail, 1) the failed hypospadias repairs are often associated with penile skin loss and deficient local tissues, 2) BMG are often not possible due to oral complications from a prior procedure, 3) patients undergoing failed hypospadias often have long urethral defects, and 4) the buccal mucosa grafts are not available due to frequent tissue harvesting. If buccal mucosa grafts are confronted with scarred tissues, BMG urethroplasty cannot be chosen again. So if surgeons are confronted with these problems in pre-pubertal boys, there is still a reliable graft to fix the poor repair.It should be noted that compared to the LMG used in adults, the advantages of LMG used in children before puberty are that the tissue material is soft and thick in mucosal tissues with a better blood supply, high cell activity and survival rate. However, there are also disadvantages in that the tongues of children have a smaller volume and are harder to secure during the procedure than tongues in adults.The length of the lingual graft in children is long enough for the defect in general, and if the urethral defect is longer, we could choose the two sides of the ventro-lateral surface of the tongue. The thickness and the articulation are related to the complications based on the outcome and the long time follow up in children. We have the relatively shorter follow up time with a good outcome, and maybe it’s a shortage of our research.The limitations of our study include 1) a short follow-up period. A short follow-up period may have had an effect on the complication rates. We had The lingual mucosal graft and the buccal mucosa graft both achieved good outcomes, and the lingual mucosa graft made up for the shortcomings of the buccal mucosa graft and provided a reliable way to treat multiple failed hypospadias procedures in pre-pubertal boys.S1 File(PDF)Click here for additional data file.


Retrograded starch is known to be resistant to digestion. We used enzyme kinetic experiments to examine how retrogradation of starch affects amylolysis catalysed by porcine pancreatic amylase. Parallel studies employing differential scanning calorimetry, infra red spectroscopy, X-ray diffraction and NMR spectroscopy were performed to monitor changes in supramolecular structure of gelatinised starch as it becomes retrograded. The total digestible starch and the catalytic efficiency of amylase were both decreased with increasing evidence of retrogradation. A purified sample of retrograded high amylose starch inhibited amylase directly. These new findings demonstrate that amylase binds to retrograded starch. Therefore consumption of retrograded starch may not only be beneficial to health through depletion of total digestible starch, and therefore the metabolisable energy, but may also slow the rate of intestinal digestion through direct inhibition of α-amylase. Such physiological effects have important implications for the prevention and management of type 2 diabetes and cardiovascular disease. Such apIn the human diet, the main source of digestible starch is that present in foods derived from cereal crops such as wheat, rice and maize, from root crops including potato and yams and also from legumes such as beans and peas. The first stage in the digestion of starch involves hydrolysis catalysed by salivary and pancreatic α-amylases to produce maltose, maltotriose and α-limit dextrins predominantly . These sThe variations in glycaemic responses are mainly attributed to differences in the rate and extent of amylolysis of different starches . The supin vitro and in vivo   . We now 22.1Wheat  and pea starches were gifts from Prof. C. Hedley and Prof. T. Bogracheva . Potato starch was obtained from the National Starch and Chemical Company  and starches from maize, waxy maize and high amylose maize (Gelose 80) were gifts from Dr. C. Pelkman, Ingredion UK .Phosphate buffered saline (PBS) tablets were purchased from Oxoid Ltd.  and PPA (type 1A DFP treated) was obtained from Sigma-Aldrich Company Ltd.  and checked for purity by SDS-PAGE. The suppliers quoted activity of 1333 units per mg of protein is equivalent to 1293 IU per mg at 25 °C 2.2g for 15 min before the supernatant was discarded and the weight recorded  of all native starches listed above were prepared in PBS and agitated in a glass beaker on a magnetic hotplate for 20 min at room temperature (∼22 °C). Aliquots from the stock suspensions were then diluted into separate 15 ml Falcon™ tubes with PBS to provide the desired range of 0.5–10 mg/ml final concentrations of starch in a volume of 4 ml.Gelatinised starch was prepared by heating sealed flasks containing 10 mg/ml stock suspensions in water at 90 °C. A thermocouple probe was used to monitor the temperature of the heating bath. Once the target temperature was reached the suspension was held for 20 min to allow for complete gelatinisation. All starches were treated in this way except high amylose maize which was pre-gelatinised at 100 °C for 10 min before autoclaving at 121 °C for 20 min to complete the gelatinisation. The pre-gelatinisation step ensures that starch sedimentation does not occur during autoclaving. All gelatinised starch suspensions were weighed before and after heating to check for loss of water by evaporation.For retrograded starch, 10 mg/ml suspensions in distilled water were gelatinised as described above, and then 10–12 ml aliquots were placed into 15 ml Falcon™ tubes and set aside at room temperature for various periods of time from 6 to 96 h. At the end of the storage times, suitable volumes of the samples were diluted to 4 ml with PBS to achieve a range of concentrations required for further experiments. Because the rate of retrogradation can be concentration dependent , appropr2.4® spectroscope equipped with a SensIR technologies IR II Durascope® diamond cell attenuated reflectance (ATR) device with a diamond crystal providing an incidence angle of 45° , peak (Tp) and conclusion (Tc) were obtained for all starches. For gelatinised starches, DSC was used to examine each sample after a cooling period at 20 °C of 30 min only, so as to minimise retrogradation. For retrograded material, two starch suspensions were prepared namely, 50 mg/ml and 200 mg/ml of distilled water. The 50 mg/ml samples were gelatinised by heating in the DSC instrument and then the ampoules containing these samples were stored at room temperature for 48 h to produce retrograded material. This retrograded starch was then heated from 20 to 150 °C at a rate of 0.5 °C per min and thermograms recorded. Comparable experiments were performed with starch samples of 200 mg per g water in order to obtain larger heat flow signals in the steel ampoules, but after gelatinisation, the ampoules were stored for 1 week at 4 °C before being re-scanned.DSC thermograms were obtained for a number of starches including high amylose maize (native and retrograded) using a TA instruments Multi-Cell Differential Scanning Calorimeter (MC DSC). Weighed samples of native starch (approximately 50 mg) in 1 ml of water were placed in steel ampoules and heated at a rate of 0.5 °C/min  while sc2.6© software (available online at http://www.originlab.com).X-ray diffraction patterns were recorded for high amylose maize starch (HAMS) and retrograded high amylose maize starch (RHAMS) using a Rigaku Miniflex 600 instrument with CuKα radiation (λ = 1.548 Å) at 40 kV and 15 mA. A small quantity of starch powder was mounted on an aluminium sample holder and levelled with a glass slide. The samples were scanned over an angular range (2Ɵ) of 3–40° and analysed using OriginPro 9.12.713C CP/MAS.NMR spectroscopy (Samples of native and retrograded HAMS were examined in the solid state by troscopy  and the troscopy .2.8A suspension in distilled water of high amylose maize starch (10 mg/ml) was gelatinised at 100 °C for 10 min prior to autoclaving at 121 °C for 20 min. The treated samples were stored at 4 °C for 24 h followed by storage at 37 °C for another 24 h . This 4 To check the quality of the purified RHAMS, the chemical composition was determined as described above for the raw starches. The mean values (w/w) ± s.e.m. obtained for the amylose, water and protein contents of RHAMS were 94.2 ± 1.1%, 3.5. ± 0.3% and 0.65 ± 0.03% respectively. The low water content was expected because of freeze drying during the preparation.2.92CO3 solution to quench the reaction. After centrifugation and collection of the supernatant, the total reducing sugar released during amylolysis was assayed by a Prussian blue method . Non-linear regression fits to the Michaelis-Menten equation was undertaken using Sigmaplot 12© software. Catalytic efficiencies (CE) were obtained from kcat/Km values.Initial rates of digestion were calculated from the slopes of plots of maltose concentration against time and used for determination, by regression, of the values of 2.10A 10 mg/ml suspension of RHAMS was prepared in PBS and agitated gently for 10 min by swirling. This was diluted in PBS to give a working inhibitor concentration of 2.5 mg/ml. To achieve the concentrations of inhibitor used in the experiments, appropriate aliquots of the working suspension were added to 15 ml Falcon™ tubes containing gelatinised wheat starch at various concentrations ranging from 0.5 to 2.0 mg/ml. Inhibitor (RHAMS) concentrations varied between 0 and 1 mg/ml. The final volume of each reaction mixture was 4 ml. Amylolysis under the conditions already described was then initiated by an addition of PPA  and samples taken for assay of reducing sugar content and determinations of initial rates of reaction.2.11k, and the total amount of digestible starch, C∞.Digestibility assays at 37 °C for native starch samples of potato, wheat, pea, maize, waxy maize starches and HAMS, were performed using suspensions containing 5 mg/ml starch and 4.5 nM PPA. Aliquots were removed at various time points up to 120 min for determination of reducing sugars. Similar experiments were conducted with gelatinised starches and 24 h retrograded starch, but with the addition of 2.25 nM PPA. Digestibility curves were analysed using the log of slope (LOS) application to 1st order kinetics  to obtai3The various starches used in the study did not differ greatly in their protein and moisture content with values (w/w) ranging from 0.1 to 0.5% and 11 to 16%, respectively. The water content of RHAMS that had been freeze dried during purification was 3.5%. The amylose content ranged between 20 and 30% (w/w) for all starches except waxy maize (1.2% amylose) and our sample of HAMS (79.1% amylose). The water uptake potential was also tested for all starches and ranged from 6.6 to 10.0% (w/w) with only potato showing a relatively high value (20.6%), which can be mainly attributed to the high levels of phosphate ester groups attached to the glucose residues of the amylopectin fraction in potato starch .Hgel of 5.5 J/g). For all the other starches, apart from potato (ΔHgel of 17.5 J/g), enthalpy values were in the range of 9–12 J/g that agreed well with previous findings from our laboratory (The endothermic transition temperatures of native starches determined by DSC are shown in Supplementary Information (Table 1S). HAMS had the largest gelatinisation range (36 °C) and the lowest gelatinisation enthalpy  determined from measurements of nditions . Thus prnditions . Althoug studies , a fall es  obtained from LOS analysis of digestibility curves  of the total starch present in digestibility mixtures but on gelatinisation the fall in peak ratio was accompanied by a marked increase in C∞ to 70–90%. . ∞ commensurate with an increase in FTIR peak ratios relative to the results obtained for freshly gelatinised samples.As expected, the total amount of digestible starch , solid state NMR and DSC. e starch  . All the values are presented as the mean ± s.e.m. obtained from 3 replicates. Increases in gelatinisation temperatures accompanying retrogradation induced by temperature cycling have been reported previously /V, where V is the maximum velocity of reaction and Ki is the inhibition constant for I binding to free enzyme to form an EI dead-end complex . For example, hitherto, the effects of water-soluble NSP (soluble fibre) on lowering glycaemic responses have been largely ascribed to increases in viscosity of intestinal contents. Such rheological changes in the gut lumen can attenuate a number of physiological processes including the rate of gastric emptying and affect the rate at which digestive enzymes interact with substrates in foods and also slow the rate of transfer of products away from the food-enzyme mix . To this4∞) and in the catalytic efficiency (CE) of PPA. The decrease in C∞ is expected if retrograded starch (RS) is completely inert to amylolysis whereas a decrease in CE can reflect direct inhibition of PPA by RS.Retrogradation of starch results in a fall in the total amount of starch that can be digested (CRetrograded starch inhibited PPA by a mixed pattern of inhibition similar to that recently reported for cellulose inhibition. It seems that PPA can interact with various carbohydrate materials and that such a reaction inhibits catalytic activity. Therefore dietary consumption of retrograded starch may not only be beneficial to health through depletion of the total digestible starch, and therefore the metabolisable energy, but may also slow down the rate at which starch is digested in the intestine through direct inhibition of α-amylase. Such physiological effects have important implications for the prevention and management of type 2 diabetes.


The authors compared the oncologic outcomes of radiofrequency ablation (RFA) with robotic-assisted partial nephrectomy (RPN) for the treatment of T1 stage renal cell carcinoma (RCC).This was a retrospective data analysis of a high-volume single tertiary centre. Patients were treated with RFA or RPN following multidisciplinary decision making. Only histologically proven RCCs were included. Baseline demographics were collected, and PADUA scores of tumour features were calculated to standardize baseline anatomy. Peri-operative complications, kidney function and oncological outcomes were compared.p < 0.001). Length of stay was shorter in RFA . Post-procedure renal function decline at 30 days was significantly less in RFA . More minor complications were recorded in RPN , but local recurrence was numerically higher in RFA . Disease-free survival (DFS) was not significantly different . Increasing tumour size was an independent predictor of local recurrence .Sixty-three cases were included in each group. Baseline renal function was poorer in RFA, and 16/63 RFA patients had tumours in single kidneys compared to 1/63 RPN cases (Both RPN and RFA offer very good oncological outcomes for the treatment of T1 RCC with low peri-operative morbidity and similar oncologic outcomes. RFA demonstrated fewer peri-operative complications and better preservation of renal function, whereas RPN had an insignificantly lower local recurrence rate. RFA should be offered alongside RPN for selected cases. With increasing numbers of renal cell carcinomas (RCC) being diagnosed annually, clinicians and researchers are continuously looking for ways to evaluate and find the most optimal treatments for RCC . CurrentDespite the several promising advantages offered by thermal ablation and other minimally invasive nephron- sparing techniques, clinicians hesitate to offer it as first-line treatment due to the uncertainty relating to its long-term oncological outcomes , 4. ThisThis was a retrospective data analysis  of the RCC databases of a single tertiary centre and was approved by the Institutional Review Board for quality improvement purposes. Patients were triaged to either RFA or RPN according to the decision of the multidisciplinary meeting on the basis of tumour size, underlying comorbidities and patient preference. In our institution, patients with stage 1a renal tumours (<4 cm) are routinely offered either RFA or RPN and consented accordingly; patients with single kidneys and/or underlying comorbidities are most often offered RFA, whereas patients with larger tumours  are routinely treated with RPN or open partial nephrectomy or laparoscopic radical nephrectomy, with thermal ablation reserved for frail patients or those with underlying comorbidities. Large renal tumours treated with open partial or radical nephrectomy were not included in the present audit. In addition, only histologically proven RCCs were included in our analysis  classification was used to score the renal tumours treated by both techniques , 15. It Peri-procedural variables were retrospectively analysed for both RFA and RPN cases in order to establish whether there were any significant differences in the short- and mid- to long-term post-operative complications using the Clavien complications grading system . Other pClinical endpoints were used in conjunction with definitions applied by the National Cancer institute (NCI) and the European Association of Urology (EAU) guidelines on renal cell carcinoma , 5. Resit test. Non-parametric data were analysed using the Mann–Whitney ‘U’ test. Disease-free Survival was analysed with the Kaplan–Meier method. Survival curves for the two groups were compared using the log-rank test and corresponding hazard ratios were calculated. Cox regression analysis  was used to produce a survival function that would predict the probability of the event of interest  occurring at a given time t for given values of the predictor variables (confounders). Regression covariates included baseline tumour size, PADUA score, age and type of treatment offered (RFA vs RPN). The StatsDirect (version 2.7) and the GraphPad Prism (version 6.0) software systems were used for statistical analysis, and a p value of <0.05  was taken as statistically significant.Baseline demographics were analysed using the Chi-square test for categorical variables and Fisher’s exact test as applicable. Continuous data that showed normal distribution, such as patient age, were evaluated using p < 0.0001), and the majority of patients had an ASA score of ≤2 (p = 0.14). Mean tumour size was significantly different in the two groups (p = 0.0003). However, the PADUA score, which takes into account tumour size as well as anatomical characteristics that can affect complexity and complications of the treatment, showed no significant difference between the two groups (p = 0.69). Furthermore, the RFA group consisted of 16 cases with a single kidney as opposed to only one in the RPN group (p = 0.0002). The baseline renal function was significantly poorer in the RFA group (51.5 ± 20.0 mls/min/1.73 m2) compared to the RPN group (87.8 ± 15.1 mls/min/1.73 m2). In addition, the RFA group included two patients with documented von Hippel Lindau disease  and three cases with multiple small renal tumours . All multifocal renal tumour cases had undergone unilateral total nephrectomy previously and were treated with sequential RFA sessions for nephron sparing purposes. The majority of tumours were histologically proven clear cell carcinomas in both RFA and RPN groups . Both Creatinine (Cr) increase and Glomerular Filtration rate (GFR) decrease were higher in the RPN group in keeping with a greater insult to the kidney. Early post-procedure renal function decline at 30 days was significantly less in RFA . Pre- and post-operative renal function in the 2 groups is illustrated in Fig. p < 0.0001). Clavien complication grading system showed a similar minor complication rate with no significant difference between the two groups (Table Table p < 0.0001), because of the more recent commencement of the RPN service (Table p = 0.64). Metastases developed in four patients  after a median period of 37 months .The median follow-up for the RFA group was significantly longer than that for the RPN group, 4.0 vs. 1.5 year, respectively  in favour of RPN . On the other hand, tumour size had a significant effect on DFS with increasing size found to be an independent predictor of local recurrence . The adjusted hazard ratio (HR) derived from the Cox regression analysis is interpreted per unit of measurement, i.e. per cm of tumour size. Hence, assuming proportional hazards (Cox model), the hazard of recurrent disease would increase by 1.7 times per centimetre of original tumour size.Disease-free survival (DFS) was not significantly different between the two groups  . RecentlBy retrospectively comparing patients at a single tertiary centre, we were able to match tumour sizes, PADUA scores and comorbidity scores, and evaluate the mid- to long-term outcomes with either treatment option. Baseline demographics were well matched in both groups, but comorbid conditions were slightly more pronounced in the RFA group. Tumour size was significantly higher in the RPN group, while significantly more single kidneys were treated in the RFA group Table . Overallp < 0.0001). It is notable that two local recurrences and two late metastases were identified in the RFA group after 3 years, which was beyond the maximum follow-up period in the RPN group. This may be due to the slow growth rate of the disease and clearly indicates the need for ongoing long-term follow-up . The baselines demographic results partly reveal the current treatment preferences among clinicians and patients Table . For exaCertain strengths as well as limitations were unique to our study and are worth discussing further. Firstly, our data were retrospectively collected and analysed, raising the possibility of selection bias. Arguably, the validity of our results may have been limited by the discrepancy in follow-up length between the two groups. This was outside this study’s control, due to the later establishment of RPN in comparison to RFA. The first RFA case performed at our centre was in 2004 as opposed to 2010 for the first RPN case. However, this was partly overcome by the more regular basis at which RPN is currently performed, giving us, thus, an adequate sample size. Secondly, our tertiary centre has certain referral patterns, for example, with referring urologists sending patients specifically for RPN or local urologists referring older and frailer patients for RFA. This might not be the case in other institutions, limiting the generalizability of our results. Thirdly, there was a significant number of lost to follow-up, especially in RFA cases. There are several factors contributing to this, most of which were outside of our control . This could have undermined our previously stated results or limited our sample of cases. Moreover, despite an adequate sample size, the low number of post-operative events limited the power of our statistical analysis.A final limitation was the heterogeneous patient demographics of our two treatment groups. However, regression analysis was employed to adjust for the confounders. In addition, despite the promising oncological outcomes of both techniques and overall disease-free survival, it is worth noting that the success of both procedures is dependent on very sub-specialized training, not currently available at all tertiary training centres and requires expertise for selecting the right patient population that could potentially benefit from either procedure.On the other hand, PADUA nephrometry scores enable comparison of our results with future studies. In addition, the use of a single technique for both RPN and RFA cases, performed by the same clinical team eliminated any variability in treatment success evaluation (e.g. operator or technique dependence). Furthermore, the fact that both Urology and Interventional Radiology departments were involved in this research as well as independent healthcare professionals (not affiliated with either department) minimized bias and strengthens the reliability of our results. Despite the above limitations, our study succeeded in confirming excellent oncological outcomes in RPN cases and emphasizing similar comparable outcomes with RFA in a selected group of patients. These results come with the added advantage that a minimally invasive approach offers: shorter hospital days, lower risk of intra-operative bleeding and potential better preservation of renal function; still little is known about long-term impact in residual renal function and adaptive kidney mechanisms with either technique. Therefore, RFA serves as an additional treatment option for the experienced urologist who can carefully recommend it to appropriately selected patients with small (T1) renal masses or as a future potential adjuvant treatment alongside RPN . In lineIn conclusion, both robotic-assisted partial nephrectomy and percutaneous radiofrequency ablation offer excellent oncological outcomes for the treatment of T1 RCC with low associated peri-operative morbidity. RFA was associated with fewer peri-operative complications and better preservation of renal function, whereas RPN had an insignificantly lower local recurrence rate. RFA could be offered alongside RPN for selected cases. Prospective randomized studies and subsequent meta-analyses in different age groups, PADUA classifications and histological variants will help confirm the interchangeable use of these two methods of treatment of small renal tumours.


Many scientists attempt to publish their work in a journal with the highest possible journal impact factor (IF). Despite widespread condemnation of the use of journal IFs to assess the significance of published work, these numbers continue to be widely misused in publication, hiring, funding, and promotion decisions , 2. Many scientists attempt to publish their work in a journal with the highest possible journal impact factor (IF). Despite widespread condemnation of the use of journal IFs to assess the significance of published work, these numbers continue to be widely misused in publication, hiring, funding, and promotion decisions , 2.There are a number of problems with this approach. First of all, the journal IF is a journal-level metric, not an article-level metric, and its use to determine the impact of a single article is statistically flawed since citation distribution is skewed for all journals, with a very small number of articles driving the vast majority of citations , 4. FurtASM journals focus on publishing high-quality science that has been rigorously peer reviewed by experts and evaluated by academic editors. The primary mission of ASM is to advance microbial science. At the recent Journals Board meeting that took place during ASM Microbe 2016 in Boston, MA, the editors in chief and the ASM leadership decided to no longer advertise the IFs of ASM journals .Our goal is to avoid contributing further to the inappropriate focus on journal IFs. Although this action by itself may have little effect on a practice that is deeply entrenched in the biological sciences, we hope that removing IFs from ASM journal websites makes a statement of principle that will be emulated by other journals.


The aim of this retrospective study was to evaluate the three-year clinical outcome for ceramic-veneered zirconia fixed dental prostheses (FDPs). All patients who were treated with ceramic-veneered zirconia FDPs, in three private practices in Sweden, during the period June 2003 to April 2007 were included. Case records from 151 patients, treated with a total of 184 zirconia FDPs (692 units), were analysed for clinical data. All complications noted in the charts were registered and compared to definitions for success and survival and statistical analysis was performed using the Kaplan-Meier method and a Cox regression model. In total, 32 FDPs in 31 patients experienced some type of complication . Core fractures occurred in two (1.1%) FDPs. Two (1.1%) FDPs or 0.6% of units showed adhesive veneer fractures. Cohesive veneer fractures occurred in 10 (5.4%) FDPs (1.6% of units). The three-year cumulative success and survival rates (CSR) were 82.3% and 95.2%, respectively. Ceramic-veneered zirconia is a promising alternative to metal-ceramic FDPs, even in the posterior area. However, the higher survival rate of metal-ceramic FDPs should be noted and both dentists and patients must be aware of the risks of complications. Combined with a search for new materials with aesthetic, biocompatible, and high-strength properties, this has contributed to the increasingly frequent use of dental ceramics in general and yttria-stabilized tetragonal zirconia polycrystals  and abutments , 4. The Compared to metal-ceramic (MC) FDPs, a zirconia restoration is more aesthetical. Further, its high fracture strength, considerably higher than alumina, enables the manufacture of all-ceramic FDPs. Which may be bilayered, using a veneering ceramic, or monolithic, using only zirconia as a full-contour restoration.Zirconia restorations for dental replacements are made using CAD/CAM technology. Production is based on scan data from a scanned abutment tooth or stone die which is processed with CAD software. The computer-designed substructure is milled either from presintered zirconia blanks (soft milling) followed by densely sintering or from fully sintered blanks (hard milling) in hot isostatic pressed (HIP) zirconia. The surface of the zirconia substructure may then be veneered with silicate-based porcelain .A few studies have been published on the clinical performance of zirconia restorations. For single crowns, Groten and Huttig have presented a 2-year survival rate of 98% . For 5-yThe aim of this three-year retrospective follow-up study was to evaluate the success and survival rates of a large number of ceramic-veneered zirconia FDPs in private practices, manufactured from the same materials with uniform methods.This three-year retrospective follow-up study was essentially compiling and analysing records from all patients consecutively treated with zirconia FDPs. Single crowns and implant-supported FDPs were not included. All FDP treatments were performed between June 2003 and April 2007, by three experienced clinicians at three private practices in Sweden. The criteria for choosing zirconia FDPs were according to the zirconia manufacturer's requirements. Case records from 151 patients  were analysed. They were treated with a total of 184 zirconia FDPs, and the distribution between the three clinics was 39, 56, and 89 FDPs. The records were evaluated for two months in late 2010 by two of the authors. Factors including age, sex, number of units (abutment teeth and pontics), occluding teeth in the opposing jaw, X-ray status, type of cement used, endodontic treatment before FDP delivery, post material, and prosthetic complications such as cohesive fractures, adhesive fractures, and loss of retention were registered. All other complications during the follow-up period were also registered. The treating dentist conducted clinical follow-up examinations once a year for three years. Patients were asked to contact the clinic if they experienced any problems with their FDP or abutment teeth. The examinations consisted of a complete dental and oral hygiene assessment, including examination of radiographs, bridge stability, and a full cariological and periodontal evaluation.Of the 458 abutment teeth, 167 were root filled and 134 were treated with posts (48 with indirect technique and 86 with direct technique) before the cementation of FDP. success [A zirconia FDP that during the entire observation period remained unchanged, free from complications, and not requiring any intervention, was regarded as success . survival [A zirconia FDP that remained in situ over the full observation period and only displaying minor modifications that did or did not require intervention, was regarded assurvival . failure.A zirconia FDP that was remade due to loss of retention, fracture or extraction of an abutment tooth, fracture of core material, adhesion fracture, extensive cohesion fracture, or pain, was regarded as2, the core thickness was a minimum of 0.5 mm, and the maximum pontic span length was 15 mm. All frameworks were milled at a production center , from fully sintered, hot isostatic pressed (HIP) zirconia  were between 0.2 and 2 mm.Abutment teeth were prepared according to the manufacturer's guidelines . All FDPs were manufactured by one dental laboratory by two experienced dental technicians according to the manufacturer's instructions. Three dental scanners were used for framework design during the inclusion period . The connector dimensions were at least 9 mmzirconia . Before n = 109; 59%), a dual-curing, self-adhesive resin luting cement  was used. Two dual-curing resin luting cement types  were used for 69 (38%) and three (1.7%) of the FDPs, respectively. A resin modified glass ionomer luting cement  was used for one FDP, and for two FDPs the cement type was not registered.For most of the FDPs  were calculated according to actuarial life table techniques, and standard errors were calculated using Greenwood's formula.P < 0.05. The data was analysed by means of the Kaplan-Meier survival probability method, and graphs for survival and failure were plotted. Further, a Cox regression analysis was performed on the data for success and survival to investigate the possible influence of different variables. The variables included in the Cox regression model were operator, number of units, being anterior/posterior, maxilla/mandible, cement type, endodontic treatment in abutment tooth, presence of post, and cantilever. Statistical analysis was conducted using SAS version 9.3 . The significance level was set at This study included 151 patients with 184 FDPs. The 184 FDPs consisted of 692 units . The mean numbers of abutment teeth per FDP and pontics per FDP were 2.5 (range 1 to 6) and 1.3 (range 0 to 4), respectively. Out of 184 FDPs, 21 were cantilever FDPs, with one cantilever pontic each.n = 143; 78%) were in the posterior region and 117 FDPs (64%) were in the maxilla  experienced some type of complication . Out of Ceramic fractures occurred in 7.6% of the FDPs. Core fractures occurred in two (1.1%) of the FDPs. One core fracture was seen in a six-unit (five abutments and one pontic) anterior FDP on a bruxist patient . The secAdhesive veneer fractures were found in two (1.1%) of the FDPs or 0.6% of all units and ten (5.4%) FDPs had cohesive veneer fractures  . Four ofAccording to the definition of success, 25 (13.6%) of 184 FDPs were considered failures. The reasons were as follows: cohesive fractures of the ceramic veneer (10), adhesive fractures (2), core fractures (2), secondary caries (2), extraction of abutment teeth (2), endodontic problems (6), and minor hygiene problems related to the FDP (1). Seven of these FDPs were remade, and two were replaced with implant-supported restorations.According to the definition of survival, seven (3.8%) FDPs were considered failures. The reasons were as follows: adhesive fracture of ceramic veneer (1), cohesive fracture in combination with insufficient aesthetics and phonetic problems (1), repeated cohesive fractures (1), complications related to endodontic treatments (2), and extraction of abutment teeth (2).The complications were evenly distributed over time and occurred equally between the three dental practices. The core fractures occurred among long-span FDPs whereas the ceramic veneer fractures where found in both short- and long-span FDPs.The three-year CSR for success and survival was 82.3% and 95.2%, respectively Tables . The resP = 0.0899) was seen for the variable cement type. Additionally, a forward stepwise Cox regression was run for both success and survival definitions. A statistical significance was then found for number of units (P = 0.033) and cement type (P < 0.0001).The output of the multiple Cox regression model for success and the model for survival showed no statistically significant differences for any of the included independent variables, but a tendency towards significance  was high with respect to the three-year observation period and was affected since one dentist treated many patients from a neighbouring country and did not attend the recalls. This could also have affected the results by means of an over- or underestimation of the data. The FDPs were fabricated in one laboratory, and the material is uniform; that is, the same zirconia core material and veneering material have been used. The manufacturing processes were carried out by a limited number of operators. The study was based on data from three private practices experienced in ceramic restorations. Being a practice-based study, it has the advantage of being set in a general dental setting rather than in a university clinic . Accordin = 109; 59%) were cemented with RelyX and 15.6% of those FDPs experienced complications. Panavia F 2.0 was used for 69 (38%) FDPs and 24.6% had complications. In contrast, Lute it was used for three (1.7%) of the FDPs and two (66.7%) had complications, and FujiCEM was used for one FDP which did not have any complications. The lack of a standardized clinical protocol, which is a drawback with the retrospective study design, makes it difficult to draw any conclusions about the cement. The same should be stated regarding the ceramic chipping; without a rigorous evaluation of the FDPs and a possibility of comparing the anatomy with the baseline FDP, many chippings might pass unreported in the patient charts. An intraoral scanner and a 3D comparison software could be used to match the scans from baseline to follow-up exams, as was done by Selz et al. [The multiple Cox regression showed no statistical significance, which was expected, since there were only a few events (less than 10 events per variable). Interestingly, a tendency towards significance for the type of cement was noted. The additional stepwise Cox regression was consequently used as a supplement to the multiple regressions. The outcome of the stepwise regression showed significance for the type of cement and the number of units. It was on the other hand not possible to see which specific cement influenced the most. However, most of the FDPs (z et al. . Regardiz et al. , 34. SinWithin the limitations of this three-year retrospective follow-up study, where only patient charts were analysed, ceramic-veneered zirconia is a promising alternative to metal-ceramic FDPs, even in the posterior area. However, the higher survival rate of metal-ceramic FDPs should be noted and compared to metal-ceramic FDPs, and a higher incidence of complications should be expected. Over the three years, the most common complication was ceramic fractures. Specifically, 6.5% of FDPs had veneer fractures, and 1.1% showed core fractures. Both dentists and patients must be aware of the risks of complications.


Renee De Waal, Karen Cohen, Matthew P Fox, Kathryn Stinson, Gary Maartens, Andrew Boulle, Ehimario U Igumbor, Mary-Ann Davies.Regarding the paper ‘Changes in estimated glomerular filtration rate over time in South African HIV-1-infected patients receiving tenofovir: a retrospective cohort study’ by Journal of the International AIDS Society, Citation: De Waal et al. Journal of the International AIDS Soceity, 20:21317. http://www.jiasociety.org/index.php/jias/article/view/21317 |https://doi.org/10.7448/IAS.20.01/21317Published in There was an error in one of the eGFR estimating equations. This led to errors in Therefore: Amongst those with a baseline and subsequent eGFR available, mean eGFR change from baseline at 12 months was –4.4 mL/min (95% Cl −4.9 to −4.0), −2.3 (−2.5 to −2.1), and 2.7 (2.2 to 3.2) in those baseline eGFR ≥90 mL/min; and 11.9 mL/min (11.0 to 12.7), 14.6 (13.5 to 15.7), and 17.8 (16.5 to 19.1), in those with baseline eGFR <90 mL/min, according to the MDRD, CKD-EPI (n = 11 112), and Cockcroft-Gault (n = 9 283) equations, respectively.