Keep Moving toward Healthy Heart and Healthy Brain (KM2H2) in encouraging physical activities for the prevention of heart attack and stroke among hypertensive patients enrolled in the Community-Based Hypertension Control Program (CBHCP).To evaluate the efficacy of the program Cluster randomized controlled trial with three waves of longitudinal assessments at baseline, 3 and 6 months post intervention.Community-based and patient-centered self-care for behavioral intervention in urban settings of China.2 plus standard CBHCP care (6 centers and 232 patients) or the standard care only (6 centers and 218 patients).A total of 450 participants diagnosed with hypertension from 12 community health centers in Wuhan, China were recruited, and were randomly assigned by center to receive either KM2H2 is a behavioral intervention guided by the Transtheoretical Model, the Model of Personalized Medicine and Social Capital Theory. It consists of six intervention sessions and two booster sessions engineered in a progressive manner. The purpose is to motivate and maintain physical activities for the prevention of heart attack and stroke.KM2HHeart attack and stroke , blood pressure , and physical activity  were assessed at the individual level during the baseline, 3- and 6-month post-intervention.2 was associated with significant reductions in the incidence of heart attack  and stroke , and moderate reduction in blood pressure (-3.72mmHg in DBP and -2.92 mmHg in DBP) at 6-month post-intervention; and significant increases in physical activity at 3-  and 6-month  post-intervention, respectively.Relative to the standard care, receiving KM2H2 is efficacious to reduce the risk of heart attack and stroke among senior patients who are on anti-hypertensive medication. Findings of this study provide solid data supporting a formal phase-III trial to establish the effectiveness of KM2H2 for use in community settings for prevention.The program KM2HISRCTN12608966ISRCTN Register  The prevalence of hypertension in China was approximately 5% in the 1950s ,3. HowevAntihypertensive therapy is highly recommended and has been widely adapted as a standard measure for secondary prevention of hypertension, including prevention of heart attack and stroke –10. ClinLifestyle modification has long been recognized as an effective measure for both primary and secondary prevention of hypertension ,12,14,15It is challenging to motivate people to participate in physical activity in general population ,22,23. Tmodel of personalized medicine . During this period, have you suffered from or diagnosed by your doctor as 2. Both systolic (SBP) and diastolic blood pressure (DBP) were measured with the mercury sphygmomanometers. Following the standard protocol recommended by the American Hypertension Association, participants were asked not smoking, not drinking, no coffee or tea, and not engaging in physical activities at least for 30 minutes before assessment. During the measurement, participants were instructed to sit down and relax at least for 5 minutes before the blood pressure was taken. Blood pressure was measured by registered nurses. Mean values of two reliable measures at least 3 minutes apart were used for analysis.Blood pressure (mmHg) at rest was used as the secondary outcome to support the efficacy of KM2H2 in encouraging participants moving from physically non-active to more active and further to maintaining active. Participants were asked to self-rate their physical activities using a self-rating scale according to the definition of moderate to intense activities . This scale was developed for use in research to assess physical activities among senior hypertensive patients with the guidance of the transtheoretical model , and stroke  with Cohen’s d = 0.36, 95% CI: .A total of 44 participants (14 from the intervention group and 30 from the control group) were diagnosed with heart attack and 61 with stroke (18 from the intervention group and 43 from the control group) during the 6-month follow-up period. After considering for the design effect of multi-center randomization with repeated measure, the main effect of time and group, the covariates of age, gender, tobacco smoking and alcohol consumption, and the adherence to anti-hypertensive medication, the adjusted incidence rates of stroke and heart attack for the two groups during the follow-up period are presented in 2 in encouraging physical activity for prevention of heart attack and stroke among a sample of 450 senior hypertensive Chinese patients who are on antihypertension medication. The program was developed with guidance of an extended transtheoretical model that incorporates the model of personalized medicine and the social capital theory. It consisted of two in-house sessions, two telephone-counseling sessions and two group-meeting sessions, all of which were delivered on a weekly basis at the community settings. Physicians in the community health centers comprised the main force of the interventionists to deliver the intervention. The trial was evaluated using the advanced mixed effect modeling approach and three waves of longitudinal data. The effect size of the intervention ranged from medium to large for physical activity measures and medium for the incidence of heart attack and stroke.In this study, we reported an efficacy trial of the program KM2H2 was substantial in reducing the risk of heart attack and stroke. Although there is a lack of data on the incidence of stroke and heart attack among senior hypertensive patients in Wuhan, data from population-based studies indicate that among the Chinese adults in general, the annual incidence rate is ranged from 200-600/10000 for stroke (before 2010) [2 can be demonstrated through a formal phase-III trial, large-scale applications of this program will produce substantial effect in reducing the number of heart attack and stroke events among hypertensive patients currently totaling hundreds of millions in China.The effect of KM2Hre 2010) ,44 and 4re 2010) . Both stre 2010) . Conside2 in reducing the risk of heart attack and stroke was further supported by the impact of the intervention on reduction in blood pressure, the secondary outcome. Relative to the standard care, receiving KM2H2 is also associated with significant reductions in both systolic and diastolic blood pressure, supporting the observed reductions in stroke and heart attack. Although the amount of blood pressure reduction was relatively small, the effect was measured over a very short period of six months. Furthermore, the reduction was achieved over the effect of the standard CBHCP care, including antihypertensive drug treatment.The efficacy of KM2H2 is supported by the observed increases in physical activity, the tertiary and target outcome variables. Compared to controls, participants in the KM2H2 were more likely to move to advanced physical activity stages with higher levels of actual physical activity engagement. These findings provide strong evidence (1) supporting adequate efficacy of KM2H2 in encouraging physical activity among senior Chinese hypertensive patients and (2) demonstrating its great potentials for community-based prevention of heart attack and strokeLastly but not the least is that the effect of KM2H2 is efficacious? The first explanation we can offer is program specificity and relevance. In preparing for KM2H2, we conducted a number of pilot studies to assess the possibilities to encourage physical activity among old adults, including those suffering from hypertension. Results from the pilot studies revealed that many old adults did engage in a number of physical activities by themselves, including quick walk, jogging, running, swimming, tai-chi, and dancing, to name a few, although many of these subjects were in their 60s and several even in their 70s. However, few of these people met the criteria of regular and moderate level activity required for prevention of heart attack and stroke. In these pilot studies, we also identified a number of barriers  and facilitators  for physical activity [2.Why the program KM2Hactivity . These p2 works. We developed the program following the extended transtheoretical model. To motivate senior hypertensive patients to engage in physical activity, we devised the intervention with the first session for knowledge enhancement, considering the low level of education and the lack of knowledge of old hypertensive patients. Physical activities are nothing new at all to many people, including old adults in China; we thus used only one in-house session, Session II for physical activity training with emphasis on skill training and problem-solving. Considering the great individual differences of senior hypertensive patients, two one-on-one counseling sessions were added, guided by the model of personalized medicine. To enhance social support and to promote self-confidence, two social capital theory-based group sessions were included. These sessions provided opportunities for participants to associate with each other, to share skills and experiences, and ultimately to build a lasting relationship to enhance social capital[Theory-ground intervention offers another explanation why KM2Hl capital,37 that l capital,37,46. Tl capital. Our fin2 through more rigorous phase-III trials.There are several limitations to this study. Although the program was evaluated using a randomized design, a few baseline measures significantly differed between the two intervention arms, including cigarette smoking and marital status, both of which may have confounded the intervention. Although such differences are not uncommon for community-based trials with a cluster randomization design , and theS1 CONSORT Checklist(DOCX)Click here for additional data file.S1 Protocol(PDF)Click here for additional data file.


To introduce a model for the time evolution of active caspase-3 protein expression in albino rat lens up to 24 hours after in vivo exposure to low dose UVR in the 300 nm wavelength region (UVR-300 nm).2 UVR-300 nm for 15 minutes. At 0.5, 8, 16, and 24 hours after the UVR exposure, the exposed and contralateral not-exposed lenses were removed and processed for immunohistochemistry. The differences in the probability of active caspase-3 expression at four different time points after exposure were used to determine the time evolution of active caspase-3 expression. A logistic model was introduced for the expression of active caspase-3. The parameters for the exposed and the not exposed lenses were estimated for the observation time points.Forty Sprague-Dawley rats were unilaterally exposed in vivo to 1 kJ/mThe exposure to UVR-300 nm impacted on the parameters of the logistic model. Further, the parameters of the model varied with time after exposure to UVR-300 nm.The logistic model predicts the impact of exposure to UVR-300 nm on the spatial distribution of probability of active caspase-3 protein expression, depending on time. One eye from each animal was exposed to ultraviolet radiation. UVR in the 300 nm wavelength region was generated with a high pressure mercury arc lamp. The emerging radiation was collimated and then passed through a water filter and a double monochromator. The output radiation .In order to determine the distribution of active caspase-3 expression, the numbers of epithelial cells in each section were split into halves, assuming symmetry around the anterior pole of the lens. Each half was split into 15 segments. The average number of cells in each half of the lens was estimated to 213 cells. Each half is presented from number zero indicative of the anterior pole of the lens and number 213 indicative of the equator of the lens.The significance level and the confidence coefficient were set to 0.05 and 0.95, respectively.There was no expression of active caspase-3 at the equator in either of the exposed or the not exposed lenses. Active caspase-3 expression increased towards the anterior pole of the lens as a function of position in the lens epithelium. But the increase rate, as a function of position, was different in exposed lenses compared to contralateral not exposed lenses .The probability of expression of active caspase-3 increased exponentially from the equator through an inflection point and then decreased towards an asymptote at the anterior pole of the lens. It is thus important to include a parameter that accounts for maximum expression of active caspase-3 at the anterior pole, a parameter for the rate of increase and a parameter for the inflection point of the rate in the model.p(n), depends on n (segment number) and the parameters; the anterior pole asymptote probability of active caspase-3 expression, ρ (probability); the rate constant, κ (segment number.−1); the phase of the probability of active caspase-3 expression η (segment number), and a unit less proportionality parameter for the exponential factor, φ. (Eqn.1).On the assumption that the increase of rate of active caspase-3 expression close to the equator, and the decrease of rate of active caspase-3 expression close to the anterior pole are the same, a simplified generalized logistic model was introduced. The model implies that the probability of active caspase-3 expression, The inflection point can be calculated from the parameters estimated as indicated in Eqn.2.Probability of expression of active caspase-3 was fit as a function of segment number and is presented in figures with segment number transformed to cell number.The probability of active caspase-3 expression as a function of cell position depends on exposure to UVR-300 nm. In the observed UVR-300 nm exposed lenses, the rate of expression of active caspase-3 increased more peripherally than in the contralateral not exposed lenses. Further, the asymptote of active caspase-3 was higher in exposed lenses than in the contralateral not exposed lenses .The expression of active caspase-3 as a function of cell number also varied as a function of time after UVR-300 nm exposure in the time window 0.5–24 hours .The expression of active caspase-3 as a function of cell position and time after exposure in exposed and contralateral not exposed lenses is shown in ρ, κ, η and φ (Eqn.1) were estimated by fitting the probability of expression of active caspase-3 as a function of position in each lens. Then, the averages of the four parameters at 0.5, 8, 16 and 24 hours after the exposure, respectively, were estimated , but no variation among non exposed lenses . The estimates of the anterior pole asymptote parameters for not exposed contralateral lenses, were not analyzed with statistical inference since the exposed and the contralateral lenses originates from the same animal. A further analysis of potential contrasts among the post exposure times with orthogonal comparison An analysis of variance of the estimates of anterior pole asymptote parameters, with a fixed effect model , indicatIn addition, the analysis indicated a difference when 0.5 and 24 hours was compared to 8 and 16 hours . The estY(t), at each time point, t (Hours), was fitted to a third order polynomial model with regression, assuming a random error, ε (Eqn.3).In order to establish the time dependence of each parameter, the estimates of each parameter, By introducing the dependence of the parameters on post exposure time into Eqn.1, and assuming rotational symmetry around the axis of the crystalline lens, a video of the time dependence of active caspase-3 evolution was generated .The current study intended to develop a mathematical model for the kinetics of active caspase-3 after exposure to UVR-300 nm.In the present paper, the time evolution of active caspase-3 after exposure to UVR-300 nm was spatially resolved within the lens epithelium. We previously showed that exposure to a low dose of UVR-300 nm induces an increase in active caspase-3 expression detectable 0.5 hour after exposure. The expression of active caspase-3 continues to increase to a peak of expression at 16 hours, and then decreases at 24 hours The current model (Eqn.1) was developed after a careful examination of scatter plots of probability of active caspase-3 expression as a function of position for each UVR exposed and not exposed lens. Both the exposed and the not exposed lenses showed a similar pattern of expression. The probability of expression increased from an apparent zero level asymptote at the equator through an inflection point in the mid region of the distance between the equator and the anterior pole. Then, the probability of expression increased with a declining rate towards an asymptote at the anterior pole of the lens. This pattern of expression led us to attempt to fit the spatial distribution of probability of expression of active caspase-3 to a logistic model . The datThe model allows independent estimates of the parameters. The model implicates a very low or no expression at the equator and a saturation of expression at the anterior pole. Further the model implies that the rate of change is proportional to the position between the equator and the anterior pole and that the inflection point has a defined position in relation to the anterior pole and the equator. The finding that the absolute value of the rate constant tended to increase faster in the exposed lenses as compared to the not exposed lenses , Table 1Our observation that the phase of the distribution of active caspase-3 expression was shifted towards the anterior pole in the exposed lenses as compared to the not exposed lenses  suggestsOur finding that the anterior pole asymptote probability for caspase-3 expression decreased towards 24 hours post exposure time is consistent with previous findings Active caspase-3 is known to be a key protein in apoptosis Considering the strong epidemiological evidence that exposure to UVR is an important risk factor for cataract ij, is the sum of the population mean, a term for the fixed variation among post exposure times, αi, and a term for the random variation among animals, εj(i) (Eqn. A.1).An estimate of a parameter, xVideo S12 UVR-300 nm.Time evolution of active caspase-3 expression after in vivo exposure to 1 kJ/m(MOV)Click here for additional data file.


Jasminum grandiflorum to produce 1 kg of jasmone containing only 0.25% of AJ. However, the jasmonatos can also be produced by the microorganisms [Botryosphaeria rhodina, making it possible to produce systems which can control the parameters involved in the fermentation process, including the use of precursors in this process.Research shows that jasmonic acid (JA) and Methyl Jasmonate (MJ), compounds that participate in the metabolism's control, development and protection of plant processes , also poBotryosphaeria rhodina Kifn 3.1 were inoculated in petri dishes containing PDA culture medium, serving later to fermentation. The homogenate was inoculated in a 250 ml Erlenmeyer flask with 50 ml of media M2. The experiment was conducted using the methodology of 22 full factorial design, evaluating time ifluencia (7 and 14 days) and the addition of the precursor linolenic acid (0 and 1 µM). All experiments were performed in triplicate. After the period of each fermentation, the material was vacuum filtered and then subjected to extraction and analysis of the production of jasmonic acid. The extraction was by liquid-liquid partition using as extracting agent as ethyl acetate. The detection of the AJ extract was made by TLC (thin-layer chromatography comparative) Samples AJ standards (Sigma) were used as reference. Quantifying jasmonatos in extracts was performed by HPLC (High-performance liquid chromatography). The calculations of the factors influencing the time and addition of precursor and it's interaction were statistically analyzed by t-test at the 5% level of probability.Inventories of line Botryosphaeriarhodina.Through the analysis of the CCDC fermentations with culture medium supplemented with linolenic acid was possible to verify that all samples had jasmonatos. The addition of linoleic acid (1 µM) induced inhibitory effect on the production of jasmonatos, however, the increase in time from 7 to 14 days to produce positively influences AJ. Thus it was concluded that the conditions evaluated is not necessary to supplement the culture medium with linoleic acid and increased time of fermentation favors the production of AJ by fungus


Objective. To propose and validate a dimensional parameter, the sagittal pelvic thickness (SPT)  for the analysis of the sagittal balance of the pelvispinal unit. Methods. The parameters were analysed on standing radiographic imaging and compared for normal, low back pain, children, and spondylolysis cases. Results. Values of SPT/S1 were observed significantly higher in high grade spondylolysis populations and in children  than in normal population . A geometrical connection with the classical angular parameters validated SPT/S1. Conclusion. SPT/S1 was considered reflecting the lever arm of action of spinopelvic muscles and ligaments and describing the ability of a subject to compensate a sagittal unbalance. It was proposed as an anatomical and functional pelvic parameter. A strict relation was described between the sagittal pelvic anatomy and the sagittal shape of the spine, particularly the amount of lordosis needed for each individual. Therefore, angular parameters were recommended because they are usable disregarding the size of the subjects –5.In the same way, the distinction was established a long time ago by morphologists and paleontologists between the “pelvis in tension” of the quadrupeds and the “pelvis in pressure” characterizing the bipedalism . They weAngular and dimensional parameters were measured on 272 lateral radiographies including the pelvis, the femoral heads and the lumbar column, in standardized standing position [The first group comprised 61 healthy voluntaries (column A). Data were obtained several years ago from for original orthopaedic studies . At thisThe third group (column C) comprised 15 children X-rayed for pathologies other than vertebral, aged from 4 to 10 years. Previous studies emphasized sagittal pelvic parameters evolving until 10 years old.The fourth group comprised 49 spondylolysis cases. Forty of them were with low grade listhesis (LGL) (Meyerling's stage 1 or 2) and without any distortion of the upper sacral plate (column D1). The mean age was of 27 years . The 9 other cases were with high grade listhesis (HGL) (column D2), Meyerling's stage 3 (8 cases) and 4 (1 case). The Chairman of our Ethics Committee attested that the data collection of all included patients and healthy subjects was in agreement with the recommendation of the Institutional Review board of the institution.The pelvic incidence (PI): value of the angle between the line perpendicular to the upper plate of the first sacral vertebra (S1) at its midpoint and the line connecting this point to the femoral heads axis. The pelvic lordosis or pelvic radius-S1 angle (PR-S1): value of the angle between the sacral upper plate and the line connecting the posterior point of sacral plate to the femoral heads axis. Pelvic tilting (PT): value of the angle between the vertical and the line connecting the midpoint of S1 and the femoral heads : the distance between the midpoint of the upper plate of S1 and the middle of the femoral heads axis.The length of S1: the distance between the anterior and posterior edge of the upper plate of S1.The diameter of the femoral heads: mean value of the diameters of two femoral heads. The overhang of S1 on the femoral heads (OVS1): distance between the femoral heads and the projection of the midpoint of the upper sacral plate, here expressed relatively to the length of the upper plate of S1 . Its anterior part was extrapolated from the anterior edge of S1 and the posterior segment of the upper plate.t-test was used to investigate the significant differences between the parameters according to the clinical groups. The Spearman's correlation coefficients were reported for the relationships between parameters.The student The mean values and the standard deviations of the parameters were reported in P < 0.001) and significantly higher in the spondylolysis groups, mostly for the high grade listhesis. The comparisons were similarly significant for PR-S1.The comparisons between the values of the parameters in each group were reported in P < 0.001), but similar for adults and LGL spondylolysis group. Also, the values of femoral head diameter were significantly smaller for the spondylolysis groups than for the reference adult normal group (P < 0.001), but not according to the listhesis grade (P > 0.1). The values of the individual ratios “femoral heads diameter—length of the upper plate of S1” were significantly different between the control group and the others, except with the low grade spondylolysis (P > 0.1).Obviously, the mean length of the upper plate of S1 was significantly smaller for children and for HGL spondylolysis group than for adults , as well than for spondylolysis subjects with vertebral slip of low grade (P < 0.01) and high grade (P < 0.05). It was not significantly different between the two spondylolysis groups in spite of different values because of the small number of cases included. On the other hand, expressed relatively to the diameter of the femoral head, the “sagittal pelvic thickness” was observed significantly different only between the children and the adult reference group (P < 0.001), and (but lower) the low-grade spondylolysis group (P < 0.05). The difference was not observed significant neither between the normal group and the spondylolysis groups nor between the two spondylolysis groups. Nevertheless, expressed relatively to the length of the upper plate of S1, the “sagittal pelvic thickness” was significantly different between the normal adult group and high grade spondylolysis group, and between the two spondylolysis groups. It was similar between adults, children, and low grade spondylolysis groups. Expressed in millimeter, the values of the “sagittal pelvic thickness” were significantly greater for the adults than for the children  between the “pelvic incidence” and the “sagittal pelvic thickness”  [At the same time that the pelvic incidence was described as the key parameter for the analysis of the sagittal balance of the spinopelvic unit, the sagittal pelvic thickness was proposed by Duval-Beaupere and colleagues to define the sagittal anatomy of the pelvis . Boulay P < 0.1) . NeverthP < 0.1) . It was P < 0.1) ) and to r = 0, 998, P < 0.001, whatever was the population group) leaded to elaborate a geometrical connection of these two angular parameters with “SPT”  and “SPT” highlighted the ability of a subject to compensate a sagittal disturbance of the spinopelvic unit. The projection of the gravity of the body segment supported by the pelvic structures  was reported to characterize the clinical condition of the sagittal balance of the spinopelvic unit. It was described to be almost vertical and usually posterior to the lumbar segment, the upper plate of S1, and the femoral heads , 18, 19 This more or less ability of the pelvis to react against sagittal disturbance was also connected to a painful torque effect of the gravity on the sacroiliac joints. In upright steady state, the gravity was projecting vertically to the axis of the sacroiliac joints  , 18, 19.As lumbar curvature was reported influencing spinal muscles , pelvic The SPT is a reliable sagittal dimensional anatomical pelvic parameter when it is expressed in value relative to the sizes of the femoral heads. It expresses the capacities of a subject to manage the sources of imbalance at the same time by its spatial compensatory adaptability and the stabilizing aptitudes of the musculoligamentous structures. SPT is strongly connected to the traditional anatomical sagittal angular parameters for the evaluation of the sagittal balance of the spinopelvic unit. Moreover, it allowed a better evaluation of the lever arms of the muscles between pelvis, spine, and hips and the sacroiliac joints. This functional analysis makes it possible to clarify the potential points of actions in revalidation and even avoid useless surgical operations. Clinical treatment should be aimed at improving the stability of the spinopelvic unit taking the sagittal anatomy of the pelvis into account and reducing mechanical stresses.


The present meta-analysis initially evaluates the reliability of dental maturation in the identification of the circumpubertal growth phases, essentially for determining treatment timing in orthodontics. A literature survey was performed using the Medline, LILACS and SciELO databases, and the Cochrane Library (2000 to 2011). Studies of the correlation between dental and cervical vertebral maturation methods were considered. The mandibular canine, the first and second premolars, and the second molar were investigated. After the selection, six articles qualified for the final analysis. The overall correlation coefficients were all significant, ranging from 0.57 to 0.73. Five of these studies suggested the use of dental maturation as an indicator of the growth phase. However, the diagnostic performance analysis uncovered limited reliability only for the identification of the pre-pubertal growth phase. The determination of dental maturity for the assessment of treatment timing in orthodontics is not recommended.The online version of this article (doi:10.1186/2196-1042-14-8) contains supplementary material, which is available to authorized users. It is well established that in growing subjects facial skeletal disharmonies, i.e. skeletal malocclusions, can be correctly treated by orthopaedic approaches. Such skeletal malocclusions include, for instance, transverse maxillary constrictions and mandibular deficiency or prognathism, which are relatively common features in certain ethnic populations , 2. HoweAs chronological age , 8 and dA further method is seen with dental maturity, which can be easily assessed through the evaluation of tooth formation , and whiHowever, a correlation analysis is not sufficient to reliably assess the diagnostic usefulness of dental maturation for the identification of the skeletal maturation phase in individual subjects. Thus, a dedicated analysis of diagnostic performance is needed. Such an analysis would need to include sensitivity, specificity, positive predictive values, and positive likelihood ratios (LHRs) , 19. IntA comprehensive meta-analysis regarding the relationship between dental and skeletal maturity is thus still missing, with a reappraisal of the diagnostic performances of previous investigations deemed necessary to definitively assess the diagnostic usefulness of dental maturation in the identification of skeletal maturity. Therefore, the present study was based on the appraisal of these missing diagnostic performance analyses in previous investigations, which were then used for the subsequent meta-analysis. In particular, studies on the maturation of the mandibular teeth in growing subjects who had never been orthodontically treated were considered.http://www.ncbi.nlm.nih.gov), Latin American and Caribbean Health Sciences , Scientific Electronic Library Online , and the Cochrane Library (http://www.thecochranelibrary.com). The survey covered the period from 1 January 1995 to 30 November 2011, with no language restrictions. The following search algorithm was used in the databases, with the asterisk symbol (*) indicating truncation:  AND ). For the search through the Cochrane Library, the whole Library  was screened with no restrictions as to the record status. Finally, a manual search was also performed by scoring the references within the studies examined and the titles of the papers published over the last 10 years in the following journals: American Journal of Orthodontics and Dentofacial Orthopedics, The Angle Orthodontist, European Journal of Orthodontics, Progress in Orthodontics, Oral Radiology, Oral Surgery Oral Medicine Oral Pathology Oral Radiology, and World Journal of Orthodontics.The present meta-analysis follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)  see Add and idenk test assuming a threshold value of 0.61 [The eligibility assessment and data collection processes were performed independently by two of the authors (GP and GHW). The data collection was carried without blinding to the authors. The intra-examiner reliability in the study selection process was assessed through the Cohen  of 0.61 . ConflicStage D. When (1) the crown formation is complete down to the cemento-enamel junction; (2) the superior border of the pulp chamber in the uniradicular teeth has a definite curved form, with it being concave towards the cervical region; the projection of the pulp horns, if present, gives an outline shaped like the top of an umbrella; and (3) the beginning of root formation is seen, in the form of a spicule.Stage E. When (1) the walls of the pulp chamber form straight lines, the continuity of which is broken by the presence of the pulp horn, which is larger than in the previous stage, and (2) the root length is less than the crown height.Stage F. When (1) the walls of the pulp chamber form a more or less isosceles triangle, with the apex ending in a funnel shape, and (2) the root length is equal to or greater than the crown height.Stage G. When the walls of the root canal are parallel and its apical end is still partially open.Stage H. When (1) the apical end of the root canal is completely closed and (2) the periodontal membrane has a uniform width around the root and the apex. For mandibular molars, the distal root is considered for staging.Use of the dental maturational method , comprisCS1. When the lower borders of the second, third, and fourth vertebrae  are flat and the bodies of C3 and C4 are trapezoid in shape. CS1 occurs at least 2 years before the pubertal growth spurt.CS2. When only the lower border of C2 is concave and the bodies of C3 and C4 are trapezoid. CS2 occurs 1 year before the growth spurt.CS3. When the lower borders of both C2 and C3 have concavities and the bodies of C3 and C4 are either trapezoid or rectangular horizontal in shape. CS3 marks the ascending portion of the growth spurt.CS4. When the lower borders of C2 to C4 have concavities and the bodies of both C3 and C4 are rectangular horizontal. CS4 marks the descending portion of the growth spurt.CS5. When the lower borders of C2 to C4 have concavities and at least one of the bodies of C3 and C4 is square. CS5 occurs 1 year after the growth spurt.CS6. When the lower borders of C2 to C4 have concavities and at least one of the bodies of C3 and C4 is rectangular vertical. CS6 occurs at least 2 years after the growth spurt.Use of the CVM method , 13, comThe studies retrieved had to correlate dental and skeletal maturity in a cross-sectional design, with an analysis of the distribution of the different maturational staging of individual mandibular teeth across the skeletal maturational stages. The methodologies used had to comply with the following two requirements:Studies that presented different maturity evaluation methods were excluded. Case reports, case series, reviews, and opinion articles were also excluded.The following data items were collected: year of publication, ethnicity, investigated teeth, sample(s) size and age, results in terms of correlations between dental and cervical maturational stages, main diagnostic indications on specific dental stages in the identification of the skeletal maturation phase, and indications on the diagnostic usefulness of dental maturity in identification of skeletal maturity Table . If a stAdequacy of sample selection description based on three criteria: (1) age and gender; (2) ethnicity; (3) systemic health conditions, clearly excluding any growth or nutritional problem; (4) any further condition, i.e. use of drugs, that might alter dental and skeletal maturation; and (5) no history of orthodontic treatment Method error analysis Adequacy of statistics Previous estimate of sample size (1 point)Blinding of measurements (1 point)The methodological soundness of each article was based on a quality evaluation method that followed pre-established characteristics that were modified from other methods reported previously , 25. TheLow: total score ≤ 3 pointsMedium: total score > 4 and ≤ 6 pointsHigh: total score > 6 pointsThe quality of the studies was considered as follows:Moreover, the PRISMA statements  for the To establish the clinical performance of each dental maturation stage for the diagnosis of each CVM stage, positive LHRs were calculated . PositivFor each investigated tooth, the secondary outcomes of interest were the percentage distributions of the different maturational stages across the growth phases  and the correlation coefficients between the dental maturational stages and the CVM stages .χ2-based Q-statistic method and I2 measurement, with significance set as p < 0.1; however, because of the moderate insensitivity of the Q statistic [I2 value ≥25% was considered associated to a significant heterogeneity among the studies [The data were combined for meta-analysis using statistical software . Heterogeneity was assessed using the tatistic , only an studies . Upon th studies . PositivOf the 337 papers retrieved by the automatic and manual searches, six studies , 28–31  was reported in only one study .The distributions of the dental maturational stages among the different growth phases for each investigated tooth are given in Tables p < 0.001, at least). In particular, when pooling the male and female subjects, the overall correlation coefficients for the canine, the first and second premolar, and the second molar were 0.57, 0.62, 0.62, and 0.73, respectively. The heterogeneity among the studies was generally proportional to the correlation coefficients, with I2 values from 81% to 97% for the canine and second molar, respectively. This heterogeneity was also similar between the sexes, with the exception for the canine, in which a greater I2 of 89% was seen for female subjects as compared to that of 59% seen for male subjects.The degrees of correlation between the dental and skeletal maturational stages according to the sexes for each investigated tooth are shown in Figure A total of 227 positive LHRs were calculated. Only 19 (8.4%) of these positive LHRs were ≥10.0, with 18 related to the pre-pubertal growth phase  and only 1 related to the post-pubertal growth phase (the second molar). Moreover, a total of 48 overall positive LHRs were obtained in the meta-analysis. Only 4 (8.3%) of these overall positive LHRs were ≥10.0, with all related to the pre-pubertal growth phase . For the pubertal growth phase, the positive LHRs retrieved in each study were generally below 2.0, with the greatest value of 9.1 seen for the second molar stage G in one study . HoweverThe present meta-analysis has reappraised the diagnostic performances of the maturation stages of four mandibular teeth for the identification of the circumpubertal growth phase in individual subjects. The data show that in spite of the high correlation coefficients seen, according to which dental maturation has been proposed as a reliable indicator of skeletal maturity Table , the diaOnly studies that scored dental maturation according to the method described by Demirjian et al  were incThe main limitations of the studies included that were judged to be of low and medium quality were the lack of a full analysis of the diagnostic performance and of blinding for the measurements , 28–31. On the basis of the distribution of the different dental maturational stages across the CVM stages and their corresponding correlation coefficients, five , 28–31 oI2 generally above 50%, not shown). This shows that the dental formation follows differential timing with regard to the growth phases [I2 up to 97%). In terms of the correlation coefficients, differences were seen between the sexes within the same tooth and study.The percentage distributions of the dental maturational stages across the different growth phases Tables , and 6 wh phases , 10 amonAs the main goal of the present meta-analysis, an appraisal of the diagnostic performances of the dental maturational stages in the identification of the growth phase was performed by calculation of the positive LHRs from the previously reported data. According to this appraisal, in only a few cases were positive LHRs retrieved in each of the studies that were above the required threshold for satisfactory performance Tables , and 10.per se can be considered as a reliable indicator of skeletal maturation; moreover, the great agreement between the results obtained from the studies included  makes the present conclusions reliable.Finally, even though the present meta-analysis did not include the radiographic hand-and-wrist maturational method, the CVM method In consideration of the diagnostic performance analysis presented here, dental maturity is not a reliable indicator of the growth phase in individual subjects. The present meta-analysis has thus revealed that the conclusions reported in previous studies were not actually supported by the results obtained in those studies , 28–31. Dental maturity and skeletal maturity are significantly correlated, although there are differences across ethnic populations.In spite of these correlations, the diagnostic performance of dental maturity for the identification of growth phases, and especially of the pubertal growth spurt, is very limited.The determination of dental maturity for the assessment of treatment timing for skeletal malocclusion is not recommended.The present meta-analysis has the following conclusions:PRISMA checklist. A document showing the PRISMA checklist. (DOC 67 KB)Additional file 1:


Currently, the use of multispiral cardiac X-ray and MRI tomography techniques has been accepted as the most promising approaches both in location and prediction of substrate of supraventricular arrhythmias both in coronary and non-coronary patients.Twenty persons  with recent (< 4 months) myocardial infarction of left ventricle were included. Paramagnetic contrast-enhanced T1-weighted SE  MRI of the heart has been employed for visual detection of risk as well of anatomic locations of atrial arrhythmias in twenty patients with previous myocardial infarction. Contrast enhancement was carried out using intravenous injection of widely available extracellular paramagnetics in a dose as high as 2 ml of 0,5 M solution per 10 kg of body mass (BM). Two groups were separated in accordance with the electrophysiologic and clinical data. The Group 1 included eight persons without supraventricular tachycardias (SVT) in the course of follow-up for four months, and the Group 2 comprised twelve patients with frequent SVT paroxysms, all were later suggested for maze procedure. Also eight persons were recruited to the control group, all without cardiac pathology.In the persons of the control group the index of enhancement (IE) of the T1-w. SE MRI of the atrium was essentially uniform over the atrial wall and did not exceed 1,03±0,4 anywhere.3, whereas 91,3±10,0 in the Group 2, p<0,02). The IE, calculated for the site of most prominent uptake of contrast, also differed significantly , as well as the overall volume of contrast uptake . When anatomically placing electrophysiologic data over the map of uptake of paramagnetic over the atrium, the electrophysiologic substrate of the arrhythmia was located in ten cases of twelve closely to the region of most intensive uptake of paramagnetic to the atrial wall.The MRI data delivered highly significant differences between the groups in average values of atrial volume  with concomitant increase of left atrial volume over 75 ml provide a prognostic factor for the nearest future manifestation of atrial tachyarrhythmias and endocardial destruction procedure.Therefore the contrast-enhanced MRI of the heart can be routinely employed as effective tool of selection of persons with high risk of supraventricular tachyarrhythmias among patients with ischaemic heart disease and previous myocardial infarction.N/A.


MADPET-4 is a high resolution PET insert under development for use in a 7-T MR. To fully exploit the capabilities of the insert, a good understanding of the physical interactions, which take place in the active and passive components of the insert, is necessary. The goal of this study was to investigate the effects of different physical interactions in an accurate model of MADPET-4 using Monte Carlo (MC) simulations. The main focus of the study was on the impact of the different active and passive components of the system on the amount of random and scattered events, including scattering in the passive components and between the crystals. The influence of the low energy threshold (50-350 keV) and different geometrical conditions in the coincidence sorting process was of particular interest. The effect of including triple coincidences was also considered in the present study. Results showed that the maximum sensitivity achieved at the minimum energy threshold (50 keV), using a geometrical condition of 4-9 sectors, was ~3.43% without passive components and dropping to ~1.36% including the passive components. Interestingly, the minimum noise ratio was obtained with a geometrical condition of 9 sectors difference at an energy threshold of 50 keV. Including triple coincidences showed no benefit for energy thresholds above 250 keV. However, low energy thresholds provided a relative gain in sensitivity, reaching 15-20% for 200 keV and a maximum of 30-40% at 50 keV.


The application of the Shuttle Walking Test (SWT) to assess cardiorespiratory fitness and the intensity of this test in healthy participants has rarely been studied. This study aimed to assess and correlate the cardiorespiratory responses of the SWT with the cardiopulmonary exercise testing (CEPT) and to develop a regression equation for the prediction of peak oxygen uptake (VO2 peak) in healthy sedentary adult men.In the first stage of this study, 12 participants underwent the SWT and the CEPT on a treadmill. In the second stage, 53 participants underwent the SWT twice. In both phases, the VO2 peak, respiratory exchange ratio (R), and heart rate (HR) were evaluated.Similar results in VO2 peak (P>0.05), R peak (P>0.05) and predicted maximum HR (P>0.05) were obtained between the SWT and CEPT. Both tests showed strong and significant correlations of VO2 peak  and R peak , as well as the agreement of these measurements by Bland-Altman analysis. Body mass index and gait speed were the variables that explained 40.6%  of the variance in VO2 peak. The results obtained by the equation were compared with the values obtained by the gas analyzer and no significant difference between them (P>0.05) was found.The SWT produced maximal cardiorespiratory responses comparable to the CEPT, and the developed equation showed viability for the prediction of VO2 peak in healthy sedentary men. The aersonnel . Thus, tAmong field tests, we emphasize the Shuttle Walking Test (SWT), which is a test of incremental structure and symptom-limited, with the pace dictated externally . The incmax) achieved at the end of the test, the results of these studies have shown differences in maximal and submaximal rates. Compared with the VO2 peak values achieved during the SWT, the predicted values of this measurement in CEPT appear to be similar, but such responses must be confirmed with the completion of CEPT since this comparison was performed only with the VO2 peak values estimated for the population studied [2 peak in healthy sedentary young and middle-aged men. We hypothesized that the SWT would promote maximal cardiorespiratory responses comparable to the CEPT, and the reference equation would be viable for the prediction of VO2 peak in healthy sedentary men.In recent years, the application of the SWT to assess the CRF of healthy individuals has been implemented –10; howe studied . Further2); and to currently not be using corticosteroid medications. Moreover, since the SWT was developed for cardiopulmonary patients, i.e., with low CRF, only sedentary subjects . All participants gave written, informed consent.This study involved healthy male participants aged 18–45 years old that were recruited from the local community. To be included in the study, the participants had to meet the following criteria: self-report of no present acute or chronic diseases; be eutrophic according to the body mass index  greater than 210 mm Hg; diastolic blood pressure greater than 120 mm Hg; sustained decrease in SBP; angina; dyspnea; cyanosis; nausea; dizziness; or by the request of the volunteer .In the second stage, 53 volunteers went to the laboratory at two different days. On the first day, the body composition measurements were obtained as described in the first stage. On the second day, the participants went to the laboratory for two SWTs, with an interval of 30 minutes between them. Completion of two SWTs with this interval had been suggested to reduce the effects of the learning test ,17. For 2), respiratory quotient (R) and HR breath-by-breath were monitored. The data was filtered and was defined as VO2 peak and R peak with the highest value obtained from the arithmetic mean of the log intervals of 30s and maximum HR (HRmax) as the highest HR value recorded during the test [max was calculated by the equation HRmax = 220 – age [During the tests of the two stages of this study, the exhaled gases were collected using a gas analyzer via the portable telemetry system . Among other variables, oxygen uptake  was compared with the VO2 peak predicted by the reference equation.To validate the equation, a different group of healthy males, composed of 20 individuals, was selected according to the same inclusion criteria of the study. This group completed the SWT as described in the preceding stages. Likewise, the VO2 peak obtained using the gas analyzer. Moreover, the validation of the reference equation was evaluated in an additional group of 20 volunteers: the values of VO2 peak obtained by the reference equation were compared with the measured values of VO2 peak obtained by the gas analyzer using the paired t-test. The level of statistical significance was P<0.05.The statistical analysis was performed using the statistical packages SPSS 20.0  and GraphPad Prism 4 . In the first stage, the normality of data was checked by the Shapiro-Wilk test and the differences among measured variables were determined by paired-t-test for variables with normal distribution or the Wilcoxon test for variables with non-normal distribution. Pearson’s coefficient of correlation was performed to study the correlation between variables and the agreement between tests was assessed by Bland-Altman analysis. The sample size was calculated based on the study by Singh et al.  and was 2 and body fat percentage of 12.0 ± 5.5%, participated in the first stage of the study. The RPE and cardiorespiratory responses obtained at the end of the SWT and CEPT are presented in 2 peak, R peak, and predicted HRmax were found. The RPE at the end of the test was significantly higher in the CEPT. Strong and significant correlations in VO2 peak  and R peak  were found between the tests. The Bland-Altman analysis also showed agreement between the results for VO2 peak and R peak on the SWT and CEPT , gait speed  and BMI . There was no significant correlation with age . A model of stepwise linear multiple regressions showed that BMI and gait speed explained 40.6%  of the variance in VO2 peak. There was no interaction of the VO2 peak with distance walked and age. The 95% Confidence Interval for unstandardized coefficients were the constants (15.149–55.514), BMI (-1.606 to -0.528) and gait speed (4.746–16.950). The reference equation for the VO2 peak in the SWT was:The characteristics of the 53 participants that participated in the second stage of the study are shown in 2 peak with the values obtained by the gas analyzer, showed no significant difference between them .The results obtained by the equation of VO2 and body fat percentage of 10.5 ± 4.9%. When the reference equation was applied in this group, the VO2 peak obtained using the gold-standard method was not statistically different from the VO2 peak predicted by the elaborated equation .The characteristics of the other group composed of 20 healthy sedentary adult men were: age 23.7 ± 3.5 years old, weight of 69.9 ± 9.2 kg, height of 1.76 ± 0.07 m, BMI of 22.7 ± 2.1 kg/mmax and of the R peak, thus classifying the SWT as a high-intensity test (maximum) since the values of the HRmax ≥ 90% and of the R peak ≥ 1:10 indicate a maximal test [2 peak and R peak and the agreement of these variables were obtained by both tests. Similar results have been observed with the values of the VO2 peak on the SWT and with the values of the VO2 peak on the CEPT performed by patients with COPD [This was the first study to compare the cardiorespiratory responses between the SWT with CEPT in healthy sedentary participants. During the SWT, the participants reached the values of the HRmal test ,19. Despmal test . Strong ith COPD ,7, idiopith COPD  and hearith COPD ,22. Thesmax at the end of the test, which might indicate the SWT to be a submaximal test. Different results of other studies in which participants of both sexes, with similar ages, have also been observed and the studies showed values of predicted HRmax ranging from 94% [max, however, the sample consisted of physically active participants, recruited from a gym, who performed the SWT at 12 levels. Because the participants were young, healthy, and physically active, the use of the SWT at the 12 levels might have generated insufficient overloading for this sample; thus, the results might have been underestimated. From the data on the walking distance in the present study, we confirmed the necessity of using the protocol with 15 levels for our sample, which consisted of healthy sedentary young and middle-aged adults, since the average distance walked exceeded the maximum distance of the 12 levels. To date, no studies have compared the cardiorespiratory responses of the SWT with CEPT in healthy participants. Given the information above, and based on the data of the present study, we suggest that the SWT provides valid measurement and assessment of CRF in healthy sedentary participants, and as in cardiopulmonary patients, the SWT in young and middle-aged people could be considered a field test of maximal effort.Due to its recent application in healthy participants, the determination of intensity of the SWT remains controversial in this population. Dourado and Guerra  and Jurgfrom 94%  to 99% [from 94% , thus infrom 94% , althougfrom 94% , young a2 peak. Gait speed as a major determinant of VO2 peak was also observed in recent studies that developed reference equations for the prediction of VO2 peak during the application of the SWT in healthy, middle-aged, and elderly participants [2 peak in the Shuttle Run Test, a precursor test of the SWT, was also determined by gait speed [2 peak. The correlation between the two measurements was negative, which can be explained by an inverse linear relationship between body composition and functional capacity. The influence of body composition on VO2 peak and the distance walked on the SWT has been reported in the literature [2 peak. As the gait speed recorded was equivalent to that of the last completed level, this finding showed a strong correlation with the distance walked and thus had a greater influence on VO2 peak in stepwise linear multiple regression analysis. Similar to our findings, age did not influence VO2 peak in young adults who underwent the Shuttle Run Test [The reference equation developed in this study explained 40.6% of the variance in VOicipants ,13. Amonit speed . As withterature ,11,17. ARun Test . DespiteRun Test .2 peak values obtained by the equation and the values of VO2 peak obtained by the gas analyzer were similar, thus demonstrating the feasibility of applying the equation to healthy, sedentary young and middle-aged men. It is noteworthy that reference equations for distance walked during the SWT in healthy participants with variances of this magnitude have been published in the literature [2 peak could not be evaluated, the reference equation elaborated has applicability only in men.Although the equation developed in this study could be explained by moderate variance, the VOterature . Lastly,2 peak in this population.It is concluded that in healthy, sedentary young and middle-aged men, the SWT produced results similar to those of the CEPT with regards to cardiorespiratory responses, and the reference equation demonstrated viability for the prediction of VO


MN Intact (n = 4), PCL-Only (n = 3), and PCL+MSC (n = 3). Ten weeks before mapping procedures in animals from PCL-Only and PCL+MSC groups, animal were subjected to MN transection with removal of a 4-mm-long segment, immediately followed by suturing a PCL conduit to the nerve stumps with (PCL+MSC group) or without (PCL-Only group) injection of MSC into the conduit. After mapping the representation of the MN in S1, animals had a segment of the regenerated nerve processed for light and transmission electron microscopy. For histomorphometric analysis of the nerve segment, sample size was increased to five animals per experimental group. The PCL+MSC group presented a higher number of myelinated fibers and a larger cortical representation of MN inputs in S1  than the PCL-Only group . In conclusion, MSC-based therapy associated with PCL conduits can improve MN regeneration. This treatment seems to rescue the nerve representation in S1, thus minimizing the stabilization of new representations of adjacent body parts in regions previously responsive to the MN.We studied the morphology and the cortical representation of the median nerve (MN), 10 weeks after a transection immediately followed by treatment with tubulization using a polycaprolactone (PCL) conduit with or without bone marrow-derived mesenchymal stem cell (MSC) transplant. In order to characterize the cutaneous representation of MN inputs in primary somatosensory cortex (S1), electrophysiological cortical mapping of the somatosensory representation of the forepaw and adjacent body parts was performed after acute lesion of all brachial plexus nerves, except for the MN. This was performed in ten adult male Wistar rats randomly assigned in three groups:  After trauma to the peripheral nervous system, return to full function is seldom achieved, particularly following nerve transection , or worsin vitro  conduit is biodegradable and biocompatible . Also, tin vitro  and surv conduit on regeneration after MN transection in mice , but theIt is well known that peripheral nerves of the brachial plexus, like the median, ulnar, and radial nerves, are responsible for carrying information from cutaneous receptors of the forepaw to the CNS. In the rat, The remarkable ability of S1 to reorganize itself in response to peripheral lesions has been widely documented . In primIn this study, we used S1 responsiveness and plasticity as parameters for accessing the functional recovery of a lesioned MN. In order to identity the region of S1 receiving inputs from the MN, all other brachial plexus nerves but the MN were sectioned immediately before electrophysiological mapping. This procedure ensured that cortical responses to forepaw stimulation could only be due to activation of MN fibers. In animals from two additional experimental groups, we removed a segment of the MN and immediately treated this lesion by suturing the stumps to a PCL tube containing or not MSC 10 weeks before mapping S1. Like in the first experimental group, S1 mapping was performed only after acute deprivation of inputs from all brachial plexus nerves with the exception of the MN.Table 1). Another four animals were used only for increasing the sample for the morphological study of the MN. The three remaining animals were used for the sole purpose of harvesting MSC from the bone marrow.A total of 17 adult male Wistar rats were used in this study. Ten of these animals were submitted to electrophysiological mapping procedures as part of the three different experimental groups described below  in accordance to Brazilian Law and the National Institutes of Health guidelines.MN Intact , served as a control for the other two experimental groups. In the MN Intact group, we mapped the cortical representation of inputs carried by the MN after its functional isolation by sectioning all other brachial plexus nerves immediately before the mapping session. In this experimental group, the same cortical region was mapped a second time, during the same experimental session, right after sectioning the MN itself. This allowed us to investigate S1 immediate plasticity after deprivation of the MN inputs.Our first experimental group, called PCL-Only) and third (PCL+MSC) experimental groups, animals underwent resection of a segment of the MN, 10 weeks before mapping S1. This lesion was immediately followed by microsurgical reconstruction with a PCL conduit implantation filled with Dulbecco’s Modified Eagle Medium  or with MSC in DMEM . Mapping procedures were performed under the same conditions as in MN Intact group. Thus, in order to identify, in S1, the representation of the regenerated MN, all brachial plexus nerves, with the exception of the treated MN, were sectioned immediately before mapping S1 in animals from PCL-Only and PCL+MSC groups. Mapping data from individual experiments in the three experimental groups were then compared.In the second  from femurs and tibias of donor rats and centrifuged at 2000 rpm for 5 min. Cells were seeded into 25 cmPCL-Only and PCL+MSC groups, a two-stage experiment was performed. Initially, animals were profoundly anesthetized with ketamine (100 mg/kg) and xylazine (15 mg/kg). After being exposed from the axilla to the cubital fossa, the right MN was transected at the level of the middle of the brachialis muscle. A 4-mm long segment of the nerve was then resected. The proximal stump was sutured through the epineurium to a 10-mm length PCL conduit  or DMEM only  were injected into the conduit with a Hamilton micro syringe. Finally, the distal stump was sutured to the other end of the PCL conduit, leaving an 8-mm gap between the nerve stumps.For animals belonging to the  conduit  using a MN Intact group were only submitted to the second (mapping) experimental stage.After 10 weeks, these animals underwent a second surgical intervention in which all other brachial plexus nerves with the exception of the MN were sectioned, and the cortical mantle exposed for electrophysiological recording (see below). Animals from the Animals were initially anesthetized with intramuscular injections of ketamine hydrochloride (100 mg/kg) and xylazine (15 mg/kg), and then atropine (0.2 mg/kg) and dexamethasone (0.6 mg/kg) were administered. Surgical levels of anesthesia were maintained with supplemental doses of ketamine and xylazine delivered intramuscularly. Throughout the experiment, heart and respiration rates were monitored and body temperature was maintained at 37°C.After adequate anesthetic level was obtained, brachial plexus nerves from the right side were exposed and then transected, with the exception of the MN. Then, the animal was positioned in a stereotaxic holder, the skin was cut, the temporalis muscle was retracted, and a craniotomy was made over the left parietal cortex. The exposed cortical surface was covered with silicone oil . A magnified photograph of the exposed cortical surface was taken to mark the sites where electrode penetrations were placed in relation to superficial blood vessels.S1 multiunit recordings were performed under standard mapping conditions as previously described in other studies of our group . Basicalx–y plane by increments of 300 μm. The receptive field of each recording site and the quality of the response were documented in drawings of the body surface. Recording sites with neurons unresponsive to any type of somatosensory stimulus or with neurons presenting weak cutaneous responses or exclusive responses to deep somatosensory stimuli were also documented. After mapping the representation of the forepaw and its surroundings in animals from the MN Intact group, the MN itself was cut to deprive the brain from all information originating in the forepaw. Then, the same sites were remapped and acute modifications of multi unit receptive fields previously located at the forepaw were documented (see below).The somatosensory representation of the forepaw and its surroundings were carefully mapped by inserting the electrode at different cortical surface positions separated in the After the mapping session, small DY crystals  were placed at 3 or 4 cortical sites to allow superimposition of the mapping data with histology for cytochrome oxidase (CO). The position of the DY crystals relative to mapping sites was marked in the photograph of the cortical surface.By the end of the mapping experiment, a supplemental dose of anesthesia was given. After a deep anesthesia level was achieved, the animal was transcardially perfused with 0.9% saline , followed by a fixative solution . After carefully removing the brain from the skull, the left cerebral cortex was carefully separated from subcortical structures and flattened overnight by placing it between two lightly weighted microscope slides in phosphate buffer with 30% sucrose. Serial tangential frozen sections of the cortex were made on a cryostat  at a thickness of 30 μm. Alternate histological sections were processed for CO , or leftFollowing perfusion, a 2-mm long segment of MN was harvested from the middle portion of the PCL conduit. This nerve segment was fixed by immersion in 2.5% glutaraldehyde diluted in 0.1 M cacodylate buffer (pH 7.4) for 2 h, washed in 0.1 M phosphate buffer (pH 7.4) followed by 0.1 M cacodylate buffer (pH 7.4), and post-fixed for 2 h in 1% osmium tetroxide containing 0.8% potassium ferrocyanide and 5 nM calcium chloride in 0.1 M cacodylate buffer (pH 7.4). The nerve segment was washed in 0.1 M cacodylate buffer (pH 7.4) and distilled water and stained in 1% uranyl acetate overnight, dehydrated in graded acetone, infiltrated with Poly/Bed 812 resin  and polymerized at 60°C for 48 h. Semithin cross sections (500 nm) were obtained and stained with toluidine blue for light microscopy analysis. Ultrathin cross sections (70 nm) were collected on copper grids and contrasted in uranyl acetate and lead citrate. Microscopy analysis and documentation were carried out using a Zeiss 900 transmission electron microscope operated at 80 Kv (Zeiss EM 900).The photograph of the cortical surface with the location of mapping sites and DY crystals was superimposed on drawings of CO-stained tissue sections made in a Zeiss Axioplan-2 microscope equipped with a color digital camera  and a motorized stage (Mac5000 LUDL) controlled by Neurolucida software  running on a Dell workstation. Drawings made in Neurolucida contained contours of individual barrels and barrel subfields, and position of the DY crystals as identified by fluorescence microscopic analysis. Using the DY crystals as fiducial markers, the superimposition between the photograph and the drawing of the histological section allowed the correlation between mapping sites and CO barrels. Barrel fields could then be used as anatomical markers of the representation of different body parts , againstSomatosensory maps were drawn using standard procedures. Basically, borders between different sensory representations were marked at an equal distance between two recording sites where neuronal responses to stimulation of different body parts were obtained. In our preparation, the cortical representation of inputs carried by the MN in S1 comprised the area containing sites where good responses to superficial cutaneous stimulation of the forepaw were obtained. Before measuring the area of the representation of the MN in S1 (RMN) obtained in different mapping experiments, we first chose a composite drawing from one experiment as reference, and then adjusted the size of the drawings from the other experiments, adjusting their sizes until the superimposition of the barrel rows of the postero medial barrel subfield (the large conspicuous barrels that represent the vibrissae) of all drawings accurately matched by superimposition. This procedure minimized variations due to differential tissue retraction that could be occurring in different experiments.PCL-Only and PCL+MSC experimental groups. We acquired images from these MN segments in an electron microscope at a magnification of 7000× for qualitative analysis.We performed qualitative and morphometric analysis of the 2-mm long segment of the MN extracted from the PCL conduit in animals of the For the quantitative analysis, we photographed the whole semithin cross section of the nerve under light microscopy using a 100× oil objective and the Neurolucida Virtual Slice Module . Then, we counted the total number of myelinated nerve fibers in the resulting microphotograph. Also, five systematic fields in each nerve cross section were sampled for measurement of fiber and axon diameters, g-ratio (axon diameter divided by fiber diameter) as well as myelin thickness. All measurements were made using Image J Software.t-test. The confidence interval was 95%, with an accepted alpha value of 5% (p < 0.05). The analyses were carried out using the Prism Graph Pad 4.0 statistical software.For MN histomorphometric analysis, paired inferences were made by Student’s MN Intact (n = 4), PCL-Only (n = 3), and PCL+MSC (n = 3). In all three experimental groups, all brachial plexus nerves, with the exception of the MN, were acutely sectioned immediately before mapping procedures. Under this experimental condition, we identified the cortical representation of median nerve (MN) inputs in S1, the RMN, as the region of S1 containing neurons responsive to light cutaneous stimulation of the forepaw. We mapped the RMN in 10 experiments, comprising a total of 639 mapping sites. Table 1 lists all mapping cases and the number of recording sites inside and surrounding the RMN in each individual mapping experiment reported in this study. An enlargement of the RMN was correlated with the treatment applied to the peripheral nerve, namely a higher number of myelinated fibers (MFs) in the appropriate g-ratio range was obtained in the PCL+MSC group as compared to the PCL-Only group.We designed our experiments to study both the repaired MN morphology and the representation of MN inputs in S1. In order to evaluate differences between the cortical representation of intact and regenerated nerves, and to further characterize the impact of MSC engraftment into the PCL tube, three experimental groups were studied: PCL-Only and PCL+MSC cases (see below), the repaired MN of each experimental subject was transversally sectioned at the level of the PCL conduit and prepared for histological analysis. The structure of the nerve was evaluated and the number of fibers was counted . Regenerated nerves from the PCL+MSC group  displayed a more organized cytoarchitecture than those of the PCL-Only group . In nerves from the PCL+MSC group, there were clusters of regenerating fibers surrounded by processes of perineurium-like cells forming fascicle-like clusters that were not so evident in nerves from the PCL-Only group. In the PCL-Only group, these clusters were more dispersed and presented smaller sizes than those observed in the PCL+MSC group . Electron microscopy revealed that regenerated nerves from both PCL-Only and PCL+MSC groups exhibited myelinated nerve fibers with well-defined axoplasm and well-organized myelin sheaths as well as many small non-MFs .After mapping the RMN in PCL+MSC group  than in the PCL-Only group . There was no difference between the PCL-Only and PCL+MSC groups when the percentage of MFs was compared in the different ranges of axon diameter . However, the analysis of the fiber diameter  in different diameter ranges showed that the PCL-Only group presented a higher percentage of MFs in the 3.0–3.99 μm range of fiber diameter than the PCL+MSC group .Morphometric analysis of the regenerated MN showed a significantly higher number of MFs in nerves from the PCL-Only and PCL+MSC groups presented a Gaussian distribution pattern as expected . However, the PCL+MSC group showed a tendency toward higher g-ratios than the PCL-Only group. Additionally, the PCL+MSC group presented a higher frequency of MFs in the 0.70–0.79 range than the PCL-Only group , which corresponds to the optimal g-ratio in the MN of adult rats  confirmed a bias of the PCL+MSC group toward the highest g-ratios . Frequency of g-ratios in the other ranges was not significantly different when PCL+MSC and PCL-Only groups were compared .The comparison of the g-ratio  demonstrated that both ult rats . The sigPCL-Only and PCL+MSC groups when all fibers of the sample were compared together .No difference in myelin thickness was observed between One of the main objectives of this study was to investigate whether the tubulization of a previously transected MN would reinnervate the skin, providing cutaneous inputs to S1, 10 weeks after the lesion. We mapped cutaneous inputs from MN fibers in the FBS region of area S1 and its surroundings, by acutely transecting all other brachial plexus nerves in an anesthetized preparation. In all three experimental groups, neurons in the RMN could be activated by gentle stimulation of different parts of the forepaw, including most of the glabrous and dorsal (hairy) forepaw, with the exception of the glabrous and hairy sides of the fifth digit.Figure 2 shows composite receptive fields obtained in individual mapping experiments from all three experimental groups. In Figure 2, the shaded area in each drawing of the forepaw corresponds to the region in which light cutaneous stimulation was able to evoke clear cortical responses in S1 neurons, considering all recording sites of a given experiment. These composite receptive fields can be interpreted as the part of the skin innervated by the isolated MN. We observed some variability in the size of composite receptive fields obtained in individual cases, especially when MN Intact animals were compared with those in which MNs had been transected . However, presence of neuronal responses to cutaneous stimulation of the forepaw in animals from the PCL-Only and PCL+MSC groups demonstrated that the previously lesioned MN was able to regenerate and carry inputs capable of activating cortical neurons in S1, at least in our experimental conditions.Figure 3 shows this quantification in individual mapping experiments of the MN intact and from PCL+MSC groups . In the MN intact group (n= 4 experiments), most of the 77 recording sites in the RMN (Table 1) presented neurons activated by stimulation of the glabrous skin of D1 (70% of the sites) and Th (75% of the sites), being followed by the digit one and digit two pads . Responses to stimulation of other parts of the glabrous paw, such as the digit 2 (D2: 19%) and the third digit palm pad (P3: 23%), were also observed. Digit 4 (D4: 10%), hipothenar pad (HTh: 8%), and digit 5 (D5: 0%) were the least represented parts of the glabrous forepaw. Cutaneous stimulation of regions of the dorsal (hairy) forepaw, with the exception of D5, evoked responses in 21% of the recording sites performed in the MN Intact group. After nerve transection and treatment, the region of the forepaw in which cortical neuron responses were elicited was more restricted than in the MN Intact group . More specifically, in the PCL+MSC group , neuronal responses were obtained only after stimulation of D1 (83% of the sites), D2 (43%), Th (32%), and P1 . This result in the PCL+MSC group probably reflects the natural bias of the unlesioned MN to innervate the radial side of the glabrous hand, with predominance of D1 and Th .In our preparation, the set of cortical recording sites containing neurons clearly responsive to superficial stimulation of the forepaw was defined as the RMN. In the RMN, the representation of some parts of the forepaw, like the first digit (D1) and of the thenar pad (Th) were represented in a higher number of recording sites than other parts of the forepaw like the other digits and dorsal hand. MN Intact animals the RMNs were the largest ones, ranging from 4.0 to 5.1 mm2 . In all cases of PCL-Only and PCL+MSC groups, we recorded neuronal responses to cutaneous stimulation of the glabrous skin conveyed by the regenerated MN . In the PCL-Only group, the RMN ranged from 1.2 to 2.0 mm2, corresponding to around 36% of that measured in the MN Intact group. In contrast, extension of the RMN in the PCL+MSC group corresponded to around 50% of that measured in the MN Intact group, ranging from 1.7 to 3.1 mm2. This represented a 43% increase in comparison with the PCL-Only group. Animals with higher numbers of MFs in the MN generally presented larger RMNs, especially when animals from the PCL+MSC group were compared with those of the PCL-Only group . This trend was also observed in individual cases of the PCL-Only group , but unexpectedly not when individual cases of the PCL+MSC group were compared with each other .The area mapped as the RMN was measured in individual cases revealing that the extent of the RMN varied somewhat in different experiments. As expected, in MN intact group. Unexpectedly, we were not able to find a clear topographic organization of the representation of different parts of the forepaw in the RMN. For example, in case 11-12 , most of the RMN seemed dominated by the representation of D1, Th, and P1. In some sites neurons responded to stimulation of only one of these three forepaw subregions while in other sites they responded to two of them or to all three, with no apparent topographic order . Besides, the location and size of the representation of different regions of the forepaw in the RMN varied in different experiments. For instance, in case 11-02, we observed a large representation of the dorsal (hairy) forepaw in the center of the RMN that was not observed in other cases .We additionally characterized the topography of the RMN in the Figures 4–7). We observed that inputs from the forepaw, carried by the isolated MN, activated most of the FBS . Occasionally, neurons in some sites in the FBS were not considered part of the RMN because they only responded to deep stimulation of the forepaw , or to superficial cutaneous stimulation of other body parts .After the mapping sessions, tangential sections of S1 were processed for CO, revealing barrel fields  that marMN Intact group , barrels inside the FBS were organized in four rows, corresponding to the representation of D2–D5 respectively, as demonstrated by Figure 6). The two other barrel rows corresponding respectively to D3 and D4 had neurons responsive to P3 only .Individual barrels inside the FBS were identified in cases 11-03 and 11-13. In case 11-03 from the MN Intact and PCL+MSC groups, we found neurons responsive to superficial stimulation of the forepaw (and thus being part of the RMN) located outside the boundaries of the FBS . Part of those neurons was located in a region anterior to the FBS corresponding to the motor cortex .In some cases of the Figures 5B,C site #1) or by vibrissae stimulation. Medial and lateral to the RMN, we found neuronal responses to cutaneous stimulation of the hind paw and inferior lip, respectively . Finally, we were able to map neuronal responses to vibrissae stimulation in the dysgranular cortical region previously identified as the “non-responsive zone” of rat S1 .The cortex surrounding the RMN was usually responsive to somatosensory stimulation. The region anterior to the RMN was usually activated by deep stimulation of the forepaw . Table 1 lists the number of sites containing neurons that changed their receptive fields after the lesion in three of the experiments.After mapping the RMN in the Figures 7B,D,F). However, in some recording sites in the previously defined RMN, neurons that were responsive to light cutaneous stimulation of the forepaw became responsive to the inferior lip, vibrissae, or the hind paw, depending on the location of the site. For instance, before MN transection in case 11-12 . Curiously, the representation of the inferior lip in this case seemed to have expanded not only toward the RMN, but also to the region anterior to the RMN, where neurons were previously unresponsive or responsive to deep stimulation of the forepaw . A similar expansion of the representation of the inferior lip after MN transection was also observed in case 11-04 .As expected, without the inputs from the forepaw, neurons in many sites in the RMN became unresponsive to somatosensory stimulation  thus grants a larger reversion of the expansion of the representation of the adjacent body parts, reflected in the larger RMN measured 10 weeks after the treatment.Similarly, neurons in the anterior half of the FBS processing MN inputs from D1, Th, P1, and D2 should receive subthreshold inputs carried by other peripheral nerves from adjacent body parts. Thus, after irreversible damage of the MN, neuronal responses to stimulation of the face, snout, and those parts of the forepaw innervated by the ulnar and radial nerves might become permanent in the anterior FBS, a well-known response to deafferentation that we observed after the acute MN lesion that we performed at the end of the mapping sessions in the ct group . Since, Besides characterizing the representation of inputs from the normal MN in S1, we also investigated the cortical representation of a repaired MN that was treated by the use of PCL conduits engrafted or not with MSC. Our working hypothesis was that the combined use of these two pro-regenerative strategies would optimize nerve regrowth, thus improving the reestablishment of physiological responses relayed to S1.Our results suggest that modifications in body representation induced by peripheral damage can be reversed by the nerve tubulization treatment. Indeed, PCL+MSC group tended to present larger extents of RMN as compared with the PCL-Only group. This finding can be interpreted as a direct effect of a more efficient peripheral recovery resulting in a higher number of total MFs in the PCL+MSC group as compared with the PCL-Only group. Indeed, this result confirms our previous study showing that MN repair by means of PCL conduit combined with MSC-based therapy improves nerve regeneration, leading to functional recovery following a traumatic lesion , thus promoting a larger area of cortical activation (a larger RMN) in S1, when compared with PCL treatment without MSC. Nonetheless, as demonstrated by 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.