In a cohort of 88 patients, the predictive models retrieved integrating morphometric, vascular, perfusion and metabolic parameters, demonstrated that two imaging approaches  are sufficient to identify possible residual disease after chemotherapy. These findings could be useful in selecting patients with residual disease, helping clinicians to tailor the radicality of the surgical approach.In patients with locally advanced cervical cancer, the availability of imaging techniques for accurately defining the residual tumor would be clinically relevant for selecting patients who could be offered a more tailored surgery. The novelty of this prospective study is the development of multiparametric predictive models of histopathological response using a unique data set with three imaging modalities  and max retrieved at PET/CT “baseline” and “final” examination, and (2) including high DWI signal intensity (SI) plus, ADC, and SUVmax collected at “final” evaluation : 0.80 (0.71–0.90) and 0.81 (0.72–0.90), respectively). Conclusion: The percentage variation in SUVmax in the time interval before and after completing neoadjuvant CRT, as well as DWI SI plus ADC and SUVmax obtained after completing neoadjuvant CRT, could be used to predict residual cervical cancer in LACC patients. From a cost point of view, the use of MRI and PET/CT is preferable.Purpose: This study aimed to develop predictive models for pathological residual disease after neoadjuvant chemoradiation (CRT) in locally advanced cervical cancer (LACC) by integrating parameters derived from transvaginal ultrasound, MRI and PET/CT imaging at different time points and time intervals. Methods: Patients with histologically proven LACC, stage IB2–IVA, were prospectively enrolled. For each patient, the three examinations were performed before, 2 and 5 weeks after treatment . Multivariable logistic regression models to predict complete vs. partial pathological response (pR) were developed and a cost analysis was performed. Results: Between October 2010 and June 2014, 88 patients were included. Complete or partial pR was found in 45.5% and 54.5% of patients, respectively. The two most clinically useful models in pR prediction were (1) using percentage variation of SUV The standard treatment of locally advanced cervical cancer (LACC) is exclusive chemoradiation therapy (CRT) ,2. Accormean), as well as 18F-FDG-PET/CT parameters such as maximum standardized uptake value (SUVmax), SUVmean, metabolic tumor volume (MTV), and total lesion glycolysis (TLG) [From this perspective, we performed a prospective study with the aim to analyze the predictive ability of transvaginal ultrasound examination (TUS), magnetic resonance imaging (MRI), positron emission tomography/computed tomography (PET/CT), as well as their complementary role in detecting residual disease after neoadjuvant CRT. In previous studies, we separately explored several single quantitative or semi-quantitative parameters of each individual imaging method, namely, TUS vascular indices, TUS contrast and morphological parameters, MRI tumor volume, MRI diffusion-weighted imaging signal intensity (DWI SI) and mean apparent diffusion coefficient (ADCis (TLG) ,12,13,1418F-FDG-PET/CT) obtained at different time points and time intervals.This study aimed to develop multiparametric predictive models for residual disease after neoadjuvant CRT in LACC patients by integrating morphometric, vascular, perfusion and metabolic parameters derived from three imaging methods  during the first 4 and the last 4 days of treatment and capecitabine  during the first 2 and last 2 weeks of treatment. Patients were evaluated according to the Response Evaluation Criteria for Solid Tumors (RECIST) 4–6 weeks after completion of CRT [This prospective study was approved by the Ethics Committee of Fondazione Policlinico Universitario Agostino Gemelli IRCCS–Università Cattolica del Sacro Cuore (ID P/572/CE/2010). All the subjects signed an informed consent form. All efforts were made to avoid selection bias, and consecutive eligible patients with histologically proven LACC (any histology) and Stage IB2–IVA disease  classification 2009 ) were enn of CRT . In patiAll three imaging techniques were performed approximately 3 weeks before treatment (“baseline”), after 2 weeks of treatment , and at 5 weeks after the end of treatment . The planned time-interval including the three imaging techniques was usually 3 days and did not exceed one week in any case. The percentage variation (delta) in the TUS, MRI and PET/CT parameters were evaluated for the “baseline”–“early” and “baseline”–“final” evaluations. Tumor volumes at TUS and MRI were calculated with the ellipsoid formula . Imaging analysis of each modality was performed blind to the others and to the histopathology.According to the interpretation criteria described below for each methodology, any abnormality in the cervix was interpreted as a tumoral lesion, which was subsequently correlated with the histopathology in each case.The histopathological evaluation was performed by a skilled gynecologic oncologist pathologist (G.F.Z.). At pathology, cervical residual disease was defined as: absent ; microscopic ; and macroscopic  . The res® 6.0 .To avoid interobserver variability, all ultrasound examinations were performed by the same examiner (A.C.T.), who has more than 15 years of experience in gynecologic ultrasound. The tumor characteristics were assessed with standardized techniques including 2D and 3D grayscale and power/color Doppler examination of cervical tumor volumes, and contrast-enhanced examination with infusion of SonoVue contrast agent  . A subjemean values were measured at “baseline”, “early” and “final” examination. Tumor diameters were assessed on axial and sagittal FSE T2-WI. The maximum tumor diameters (maxTD) were recorded in the three dimensions obtained in the sagittal and in the axial T2-WI. DWI images were analyzed qualitatively, referring to signal intensity of the tumor, which was classified as hyperintense or hypointense in comparison with the adjacent skeletal muscle. The ADC map was generated by using a designated workstation  and was analyzed using the Functool dynamic analysis tool . Three freehand regions of interest (ROIs) were drawn on a single DW image where the lesion diameters were maximum, using axial T2-WI as guidance. Areas of necrosis within the tumor were avoided. The ROIs were copied to the corresponding ADC map, and the mean ADC (ADCmean) was obtained. In the absence of high signal intensity on DWI, the ROI was placed on the cervical stroma, in the site of the tumor at the baseline DW-MRI. Tumor response was classified as follows: (1) complete response in patients with total restoration of the zonal anatomy of the cervix  or with areas of intermediate or high signal intensity on T2-WI and no signal intensity on DWI or high signal intensity on DWI but with an ADCmean value > 1.1 × 10−3 mm2/s, and (2) partial response in patients with residual disease based on evidence of a residual hyperintense mass within the cervix on T2-WI with evidence of signal intensity on DWI and an ADCmean value < 1.1 × 10−3 mm2/s [mean ≤ 1.1 × 10−3 mm2/s, and equal to 0 in cases of high DWI and ADCmean >1.1 × 10−3 mm2/s or in all cases with low DWI SI.Pelvic conventional and DW-MRI were performed and reviewed according to a previously described protocol using a 1.5-T superconducting magnet  . Cervica−3 mm2/s . Accordi18F-FDG injection (3 MBq/kg) and reconstructed using the line-of-response row-action maximum likelihood algorithm  [max, SUVmean, MTV and TLG were calculated using a gradient-based method  [18F-FDG uptake at the primary site higher than the surrounding background was considered abnormal and interpreted as positive. Tumor response was classified as follows: (1) complete response in patients with absence of abnormal 18F-FDG uptake at the site of the cervical tumor; (2) partial response in patients with residual abnormal 18F-FDG uptake at this site [Standard PET/CT scans (without iodine contrast) were acquired from the skull to pelvis according to a previously described protocol using 3D Gemini GXL Philips Medical Systems at 60 min (±10 min) after × 4 mm3) . The imaOH, USA) ,20. Any his site .Sample size was calculated to detect a 15% difference in the accuracy of MRI. Based on MRI diagnostic accuracy = 75% (p0), type-1 error = 0.01, and type-II error = 0.1 , a total of 86 patients would be required. Assuming a dropout rate of around 10%, a final sample size of 95 patients was planned [Clinical, pathological, and imaging characteristics were described as n (%) or median (min-max) as appropriate. For the analysis, patients were divided in two groups  according to pathological response (reference standard).The Shapiro–Wilk test was used to test normality and comparisons between the two groups were made with the Mann–Whitney U test and χ2 test as appropriate. Imaging parameters were analyzed at “baseline”, “early”, and “final” time points. Moreover, differences between “baseline”–“early”, and “baseline”–“final” time intervals were also evaluated according to the following formula:p value less than 0.05 when comparing partial versus complete pathological responders at inferential analysis were analyzed  or time interval , the selected parameters were included in univariable logistic regression analyses in order to evaluate their performance in pathological response prediction. Those parameters that showed a terature . Multivap value in the univariable analysis was included in the multivariable analysis, both for morphometric (maximum tumor diameter and tumor volume) and SUV (SUVmax and SUVmean) parameters. In the case of equal p values, the criterion used for parameter selection was the maximization of area under the curve (AUC).To avoid collinearity, for each examination, only the parameter with the lowest All estimations and AUC values were provided with 95% confidence intervals (CIs). AUC values between 0.70 and 0.80 were considered acceptable, those between 0.81–0.90 excellent, and those > 0.9 outstanding . When thp value < 0.05 was considered statistically significant. No imputation was carried out for missing data. The statistical analysis was performed by an experienced biostatistician (TP) using STATA software .A two-sided test was used and a An analysis of the costs related to the different models developed was carried out.Each examination cost was valued according to the outpatient tariff of the Lazio Region  which represents the reimbursement that the Regional Health System recognizes to the structures that perform the services. The examinations were identified according their specific regional codes.Between October 2010 and June 2014, 108 patients were initially screened. Of these, 16 refused early evaluation and two died during CRT; 90 patients completed neoadjuvant CRT and imaging studies, two of whom showed progressive disease at the final assessment and were excluded. Thus, 88 patients were included in the final analysis : 11 patiThe clinical and pathological features of the study population are summarized in Overall, 40/88 (45.5%) had pR0, while 48/88 (54.5%) patients had PR, including 21 pR1 (23.8%) and 27 pR2 (30.7%). A significant difference was found for grading of differentiation and metastatic LNs at histology, whereas borderline significance was found for histotypes. At histopathology, metastatic pelvic lymph nodes were detected in 10/88 (11.4%) patients and in all patients with a residual cervical tumor.Of 95 parameters extracted from the three imaging modalities, only 34 (36%) were eventually considered in the present study .mean for PET/CT;At “baseline”: color score and tumor peak enhancement for TUS, none for MRI , and SUVAt “early” examination: maxTD and VI for TUS, maxTD for MRI, and MTV for PET/CT;max for PET/CT;At “final” examination: no parameters for TUS, maxTD and the combined parameter of high DWI SI plus ADC for MRI, and SUVmean (%), ΔMTV (%), and ΔTLG (%) for PET/CT;For Δ “baseline”–“early” parameters: ΔTumor volume (%) for TUS, ΔTumor volume (%) for MRI, ΔSUVmean (%) for MRI, ΔSUVmax (%) for PET/CT.For Δ “baseline”–“final” parameters: none for TUS ; Δmaximum tumor diameter and ΔADCmax values evaluated at “baseline” and “final” evaluation .The model with the highest lower 95% CI limit was that considering the variation of SUVmean;Model 1, at “baseline” examination: VFI and SUVModel 2, at “early” examination: only one ultrasound parameter (vascularization index);max;Model 3, at “final” examination: high DWI SI plus ADC and SUVmean (%), ΔMTV (%), ΔTLG (%);Model 4, for Δ “baseline”–“early” parameters: ΔSUVmax (%).Model 5, for Δ “baseline”–“final” parameters: ΔSUVAll the models had an acceptable AUC with a superimposable 95% CI  and model 5  had an excellent AUC showing the highest lower 95% CI limit ≥ 0.72. Moreover, the results for model 5 were similar to the ones shown in i1 for i = 1…82) was a dichotomous parameter and SUVmax (xi2 for i = 1…82) was a continuous parameter . Conversely, a patient with high DWI SI plus ADC = 1 and SUVmax = 10.5 at “final” examination would have a 99.9% probability of having residual disease.For instance, when applying this model, a patient with high DWI SI plus ADC = 0 and SUVmax values evaluated for the time interval from “baseline” to “final” evaluation was considered the most clinically useful. The use of parameters derived from the three imaging methods showed similar results in terms of superimposable AUC for most of the evaluations and confirmed the role of ΔSUVmax in pathological response prediction for the time interval “baseline”–“final” evaluation. Moreover, another model considering high DWI SI plus ADC of MRI and SUVmax of PET/CT at “final” examination (model 3) had an excellent AUC: patients with both high DWI with ADCmean ≤ 1.1 × 10−3 mm2/s and high SUVmax detected 5 weeks after treatment are more likely to have a partial pathological response.Using multivariable analysis, this prospective study generated models including the best parameters of three imaging methods  for predicting partial pathological response after neoadjuvant CRT in LACC patients. We found that the use of multiple parameters retrieved from the same imaging method resulted in models with superimposable acceptable AUCs for before, during and after treatment. The model considering ΔSUVRegarding cost analysis, model 1 and 2, although less expensive, cannot be taken into consideration as they include ultrasonography plus color Doppler which are not considered in the main national and international clinical guidelines. Model 3, which includes both MRI and PET/CT seems the most appropriate as it provides the necessary clinical information at a lower cost than models 4 and 5, which include PET/CT at two time points. Furthermore, in the Italian context, adoption of model 3 would allow lightening the workload for PET centers, which have less availability in the national health system. A further possible advantage, for facilities equipped with an integrated MRI/PET device, is performing both exams in a single session, resulting in a more timely and comprehensive report and a single outpatient access.To our knowledge, this is the first study elaborating a predictive model of pathological partial response including the parameters of three different diagnostic methods. Indeed, other studies investigated the role of TUS, MRI and PET/CT parameters in LACC patients, but none of them evaluated the complementary role of the three methods before, during and after CRT ,24,25,26We chose to perform “early” imaging after two weeks of treatment considering that at this time, radiation-induced inflammation was supposed to be very low. We chose to perform “final” imaging 5 weeks after completion of neoadjuvant CRT, a time that was earlier than standard of care, which is 3 months after completion of exclusive CRT. In fact, it is assumed that at this time (3 months from the end of CRT), radio-induced inflammation should have resolved, with few false positive results. It must be stressed that the total RT dose used in our protocol was lower than that used with exclusive CRT as surgery substituted for utero-vaginal brachytherapy. In any case, the time we chose was a satisfactory compromise, considering the time of surgery, which was planned within 8 weeks from the end of CRT .A potential limit of the study is that we excluded from the analysis patients with no response at neoadjuvant CRT; however, the number of patients with no response was too small to be included (two patients). Second, we are aware that a more recent FIGO stage (2018) is available ,35, whicThe novelty of this study is the development of multiparametric predictive models using a unique data set with three imaging modalities, three time points, histopathological correlations, as well as a thorough consideration of different imaging parameters. Moreover, the present study provided some new findings. First, imaging performed after two weeks of treatment (“early” examination) is not so advantageous in clinical practice, regardless of the imaging procedure. Second, the predictive models demonstrated that two imaging approaches  are sufficient to identify possible residual disease after CRT. These findings could be useful in counselling patients before treatment, and above all, in selecting patients with residual disease, thereby helping clinicians to tailor the radicality of the surgical approach. From a cost point of view, the use of MRI and PET/CT at “final” evaluation is preferable.


However, these effects were temporal and most significant in the initial minutes of treatment. This indicated a dynamic bubble size distribution which stabilised after around 15 min. Defluorination rates of PFOS were compared with measured potassium iodide dosimetry, calorimetry, sonoluminescence (SL), and sonochemiluminescence (SCL). Flow rates which enhanced defluorination correlated moderately with enhanced SCL and negatively impacted SL, calorimetry, and dosimetry. Effects were attributed to perturbed cavity surfaces, leading to asymmetric cavity collapse, and the possibility of enhanced solvated electron production/interaction. SL, SCL, dosimetry, and calorimetric measurements were also temporal, and each showed different times to equilibrate. Flow rates of 439 and 889 ml min−1 returned all sonochemical measurements to the levels without flow, likely due to continued collapse temperature quenching by furthered bubble asymmetry. Flow also enhanced reactor cooling, which is significant for industrial temperature control. The pump energy consumed was small (≈1.9 %) compared to that of the amplifier and chiller, hence, PFOS defluorination was more cost-effective using flow. However, the effect may be limited for the longer treatment times needed for environmental remediation.The removal of per- and polyfluoroalkyl substance (PFAS) pollution from the environment is a globally pressing issue, due to some PFAS’ recalcitrant, bioaccumulative, and carcinogenic nature. Destruction via ultrasonic waves (sonolysis) is a promising contender for industrialisation due to; moderate power consumption, applicability to several PFAS and sample types, and limited by-products. Liquid flow rate through an ultrasonic reactor can affect the size, shape, and spatial distribution of ultrasonic cavities and hence their chemical activity. Such effects have not been studied during PFAS sonolysis, and temporal effects have not been studied much beyond the reactant concentration. Here, the effects of varying recirculating flow rate on the ultrasonic defluorination of perfluorooctane sulfonic acid (PFOS) and implications for industrial scale up are presented. Under the ultrasonic power (200 W L Ionic strength adjustor (ISA) TISAB I was purchased from Cole-Parmer. 98.3 % sodium hydroxide was purchased from Fisher Chemical. All solutions were made using distilled water (Milli-Q) from an Elix Essential 3 (UV) Type 2 device operating at 18.2 MΩ cm.2.2−1 . The reactor was filled with 400 ml of 10 mg L−1 PFOS solution, as per our previous work 2 with the liquid. Power was supplied to the transducer using an AG 1006 T&C Power Conversion amplifier. The impedances of the amplifier and transducer were matched using a low frequency RF impedance step-up transformer (SUT) operated as a step-down transformer and static was discharged through a 10 kΩ resistor, used to ground the line.The aqueous PFOS solution was sonicated in a glass jacketed 0.6 L reactor as previously described 2.3−1 aqueous PFOS was sonicated for a total sonication time of 30 min. Note that this concentration exceeds bubble saturation conditions such that minor changes in initial concentration will not affect the zero order degradation rate In all PFOS experiments, 10.0 ± 1.0 mg L2.42.4.1−1 K−1) −1).The load power supplied to the transducer was set via the amplifier and balanced against reflected power such that Equation −1 . The temperatures measured during calorimetry and the associated cooling functions are plotted in 2.4.23− production. Hence, the samples were diluted 5.0 x prior to measurement to obtain differentiable results. The I3− concentration was then calculated from Eq. (4), according to the Beer-Lambert law 3− ions  3− ions (Mols L−1)Hart and Henglein’s dosimetry method 2 which can oxidise KI Note that, in air saturated systems, this method can lead to inaccurate absolute radical concentration measurements, due to the formation of nitric acid from N2.4.3Sonoluminescence (SL) and sonochemiluminescence (SCL) light emissions were captured using an ANDOR iXon3 EMCCD camera and associated ANDOR software. The camera was placed adjacent to the reactor, inside a light-sealed box within a dark room. SL emissions were assessed by sonicating 420 ml of Milli-Q water at room temperature  while the camera was operated at −70 °C with an electron multiplying (EM) gain of 50 and exposure time of 15.0 s. SCL emissions were similarly assessed by sonicating a 420 ml solution of 0.1 M NaOH and 1.0 mM luminol using an EM gain of 4 and exposure time of 2.0 s. The SCL EM gain and exposure time were reduced compared to SL due to the greater level of SCL light emission, which would otherwise saturate the images. Additional SL/SCL images and measurements were made using a tubular glass reactor with the same internal diameter as the reactors shown in 2.4.4− ions to be assessed but is moderately affected by pH changes −6–10−1 M). The solutions were magnetically stirred at 100 rpm and temperature controlled using a heated plate at 25.0 ± 0.05 °C. A volume of ionic strength adjustor equal to 10 % of the fluoride sample was then added, and conductivity was recorded after the reading stabilised (indicated on the conductivity meter). The correlation between [F−] and solution conductivity is shown in −] using the derived correlation.The release of fluoride ions correlated well with PFOS degradation in previous work and the same device (a Cole-Parmer Combination Fluoride Ion Selective Electrode) and methodology were used here 33.1−1), flow had a moderate (up to +14 %) impact on the PFOS ultrasonic defluorination rate during 30 min of sonication  compared to fluoride release rate at 0- and 439 ml min−1. The statistical significance to the peak was not as noticeable (P < 0.15) for flow rates 79- and 889 ml min−1. Hence, an optimised flow rate could provide 14 % faster treatment times compared to non-optimised flow or batch systems. This would reduce operating times and costs at full scale and enable the development of a flow through- or recycle-system for the treatment of PFAS contaminated waters. The presence of a maximum rate suggests competition between physiochemical phenomena as the flow rate is increased and is consistent with similar prior research − release rates.Under the recirculation rates studied  with increasing volumetric flow rate, showing significant error bar overlap in . Similarlow rate  and SL dse rates . So, KI −1), and the top of the reactor becomes homogenised and grey in colour. This trend is reversed as flow rate increases beyond 214 ml min−1. Similarly, under no flow, the region parallel to the liquid inlet (highlighted with lower red dashed box) is heterogeneous with small regions of black and white “mottled” throughout. These regions of activity and inactivity are likely due to quasi-acoustic streaming (bulk liquid flow) caused by the transducer −1, these regions agglomerate into larger, less populous, regions of black and white. This trend is also reversed as the flow increases beyond 214 ml min−1. Deformation of these regions at intermediate flow rates and (re)formation at high and low flows qualitatively agrees with the trends in fluoride release rate, SL, SCL, dosimetry, and calorimetry  also confirm findings at 376 kHz, using an overhead stirrer The blurring of light and dark SCL regions has been noted previously with the addition of flow via overhead stirring −1, the mixing region moved towards the transducer centre, leading to disruption of quasi-acoustic streaming  without an equivalent rise in cooling power until 439 ml min−1. The same power was applied to the transducer in all tests, hence, the “lost” calorimetric power must have been converted into some other form of power, such as fluid motion − release rates, SCL intensity, and SCL activity/inactivity homogenisation at these flow rates.Calorimetric power varied more significantly with flow (21.2 %) than fluoride release, dosimetry, SL, or SCL , superoxide radicals (O2•−), hydrogen superoxide radicals (HO2•−), H2O2, high temperatures, and solvated electrons (e−aq) − release rates, sonothermal and sono-radical mechanisms are unlikely dominant causes for PFOS degradation. Reduced I3− formation rates at 79.2- and 214.2 ml min−1 (•− + O2•−) radical production rate 2•− formation − release rates , and likely several other species. Hence, the impact of enhanced e−aq production on any one metric would be limited, as evidence by the small SCL intensity changes. Note that SL intensity cannot be subtracted from that of SCL for this work (to observe absolute SCL changes) since the two data were recorded under different photometric conditions  rates may be related to the observed flow effects on bubble and wave dynamics  −aq production as well as PFOS and O2 exposure to the bubble core, via increased bubble asymmetry. In turn, this enhanced the frequency of PFOS and O2 interaction with e−aq, leading to both increased PFOS defluorination and SCL.The causes of altered edynamics . As discdynamics  [28]. Enmics  interaction with radicals produced. While likely limited, bulk mixing effects are discussed further in At higher flow rates, bubble collapse temperatures were sufficiently reduced, by further asymmetry and non-volatile species injection, to counter the enhanced sonochemical activity seen at 79.2–214.2 ml minion rate , SCL intion rate , and calion rate  at high ion rate  were enhogeneity , suggestogeneity . Off-cenBesides collapse intensity, flow and mixing can reduce bubble sizes, degassing, and reactor dead zones by reducing bubble coalescence via secondary Bjerknes forces 3.4− production rates  were measured every five-ten min during the 30-min reaction and averaged. However, when comparing the F− release in five-min intervals (−1 min−1 (at 214 ml min−1) and the minimum was 2.46 μmol L−1 min−1 (at 79 ml min−1), giving a variation within 14.2 % of the minimum with varied flow rate. However, within the first 5 min of treatment, a maximum rate of 2.89 μmol L−1 min−1 was observed alongside a minimum of 1.67 μmol L−1 min−1, giving 73.1 % variation from the minimum. Variations in F− release rate from the minimum in the 0–5, 5–10, 10–15, 15–20, 20–25, and 25–30-minute intervals are 73.1 %, 14.0 %, 24.8 %, 16.1 %, 8.5 %, and 15.6 %, respectively. In the first half of the reaction, the rate variation with flow is 37.3 %, while in the latter half it is 13.4 %. For ntervals A–C, diffntervals , the max−1 data  regardless of the power used −1 data C. Conver3− generation rate compared to the 10-min fluoride release rates . However, as demonstrated in The dosimetry data was taken every 10 min and the inverse of Ise rates A–C. The se rates A. Howeve−1. This suggests there is already a well homogenised bubble population which varies little with time, which agrees with the fluoride release rates  and 3.2 % change for that of SCL . Prior work demonstrated that SL intensity was quasi-stable after just 0.3 s, oscillating around a peak value −2 without flow to sonicate 1.0 mM sodium dodecyl sulfate (a surfactant), compared to continuous 410 kHz ultrasound with an intensity of 2.27 W cm−2 and flow of 0–889 ml min−1 of pure water, used here. The former work also discussed that shorter pulse off times and lower surfactant concentrations hasten the time to reach peak SL intensity 2.In The discussion on flow effects over time and the S(C)L images in − release, SCL intensity, dosimetry, calorimetry, etc.) would likely be reduced further at higher frequencies and increased at lower frequencies, as seen experimentally − release and calorimetry taking around 15 min, while dosimetry takes 20 min or more. Such times are in agreement with measured equilibrium time of an ultrasonic reactor to a step input in reactant concentration, and thus the time to reach homogeneity The limited impact of flow on several of the sonochemical metrics used here (F4−1) enhanced defluorination rates up to 14 %, while higher flow rates (439–889 ml min−1) brought the rate back to a level comparable to that without flow. The defluorination rate was then compared to several sonochemical measurements and showed correlation with SCL, but inverse correlation with calorimetry, dosimetry, and, to some extent, SL. These findings were in general agreement with prior work and indicated that flow perturbs bubble surfaces and collapse shapes from “smooth and symmetrical” to “rough and asymmetrical”. This asymmetric collapse likely enhances solvated electron production and/or the brings PFOS molecules closer to sites of electron production, thus enhancing defluorination. However, sufficiently high flows (and collapse asymmetry) likely quenched collapse temperatures and sonochemical intermediate production . The decline in radical production but enhanced SCL with the onset of flow was likely due to relative changes in HO2•_ and O2•_ radical populations. The subsequent rebound in radical production at high flows was possibly due to enhanced mixing and reduced buffering of acoustic streaming by the incoming fluid. All sonochemical effects were also time dependent with flow, indicating that bubble populations and size distributions were dynamic until a certain equilibrium time was reached. The time to equilibrate could have significant impacts for assessment of sonochemical activity, since the time of action is different for each. Hence, researchers should consider both pre- and post-equilibrium measurements when using flow in ultrasonic reactors. The effects of flow were limited due to the dynamic, well mixed nature of the sonochemical reactor, and the power and frequency used (previously optimised) which literature suggests produce limited Bjerknes forces. Since the addition of the pump scarcely affected power consumption (+1.9 %), treatment efficiency was improved with moderate flow rates. Thus, a flow through system may be economical for industrial scale treatment, as is commonly done in water treatment industries. Future work should consider waste stream specific flow optimisation and whether flow effects are similar for other PFAS and PFAS in complex matrices, which may affect acoustic streaming and hence collapse symmetry differently to pure PFOS. Higher flow rates and different flow orientations would likely also encourage different sonochemical effects.The effects of recirculating flow on the rate of PFOS defluorination by sonolysis were considered. Moderate flows (79–214 ml minThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Among these enzymes, the class B β-lactamases are members of a superfamily of metallo-β-lactamase (MβL) fold proteins which are characterised by conserved motifs  and are not only limited to bacteria. Indeed, as the result of several barriers, including low sequence similarity, default protein annotation, or untested enzymatic activity, MβL fold proteins have long been unexplored in other organisms. However, thanks to search approaches which are more sensitive compared to classical Blast analysis, such as the use of common ancestors to identify distant homologous sequences, we are now able to highlight their presence in different organisms including Bacteria, Archaea, Nanoarchaeota, Asgard, Humans, Giant viruses, and Candidate Phyla Radiation (CPR). These MβL fold proteins are multifunctional enzymes with diverse enzymatic or non-enzymatic activities of which, at least thirteen activities have been reported such as β-lactamase, ribonuclease, nuclease, glyoxalase, lactonase, phytase, ascorbic acid degradation, anti-cancer drug degradation, or membrane transport. In this review, we (i) discuss the existence of MβL fold enzymes in the different domains of life, (ii) present more suitable approaches to better investigating their homologous sequences in unsuspected sources, and (iii) report described MβL fold enzymes with demonstrated enzymatic or non-enzymatic activities. Initially discovered in bacteria due to their effectiveness against antibiotics with therapeutic interest in humans, β-lactamases are a group of enzymes capable of degrading several β-lactam antibiotics ,2. They In this review, we aim to summarise the results of this approach, which we initiated a few years ago to analyse the functions, origin, and distribution of this group of enzymes, especially metallo-β-lactamases (MβL), that have multiple more or less specialised functions depending on the ecosystem within which they are established . Here, w2+, Fe2+, Mg2+, or Ca2+) for their activity . Th. Th43]. Pandoravirus dulcis and Pandoravirus salinus. Two other gvMβLs appear to be clustered with sequences from humans and one gvMβL grouped with Asgard MβL fold sequences , we were able to identify fifteen confirmed MβL fold sequences (based on a protein length of at least 200 aa). Default protein annotations of these gvMβL sequences from the NCBI database included ribonuclease Z, the ribonuclease BN/tRNA processing enzyme, the MβL fold metallo-hydrolase/oxidoreductase superfamily, the metallo-β-lactamase superfamily protein, the Ankyrin repeat domain-containing protein, and proteins with unknown function. However, as shown in equences .Tupanvirus deep ocean (TupBlac protein), naturally involved in translation mechanisms in giant viruses. Protein analysis using a conserved domain search (CD Search) tool [E. coli BL21(DE3) strain and its enzymatic characterisation demonstrated a dual β-lactamase and ribonuclease activity, since the TupBlac protein was able to hydrolyse nitrocefin, penicillin G, and RNA substrate (of bacteria or Acanthamoeba castellanii) [To the best of our knowledge, no studies have yet reported the activities of these MβL fold enzymes from a giant virus. Thus, in our recent work, we characterised the enzymatic activities of a MβL fold protein from giant ch) tool  revealedch) tool . As descellanii)   ,70. Thisechanism . This maDespite the default annotation of these enzymes, which may suggest a hydrolytic activity on antibiotics or RNA/DNA substrates, no studies have thus far reported enzymatic characterisation of these proteins. Thus, after we reported the presence and wide distribution of the MβL fold sequences in CPR microorganisms, we experimentally expressed, purified, and tested the enzymatic activity of five selected β-lactamase proteins, including class A and class B metallo-β-lactamases from CPR, to evaluate their ability to hydrolyse various substrates including β-lactams, RNA, and DNA  Table 1Table 1. Methanosarcina barkeri) [Pseudomonas, Burkholderia, and Pandoreae [In this review, we have highlighted the considerable extension of core genes for β-lactamases into all domains of life, as shown in barkeri) , given tbarkeri) ,75. We candoreae ,77.Our recent work highlights a wide distribution and great conservation of the MβL fold proteins in archaea . By usinThe evidence of several activities including β-lactamase, nuclease, ribonuclease, lactonase, glyoxalase, phytase, and potentially other unidentified hydrolase activities from enzymes of the same family highlights the significance of accurately naming enzymes in order to understand their nuclear reactivity. This is particularly challenging due to the scarcity of competent reviewers outside the bacterial world able to evaluate knowledge extension beyond the initial enzyme activity identification, which remains extremely problematic. Classical Blast analyses present some limitations, for example, when it comes to identifying remote homologous sequences, because of the few similarities between them. This is why alternative search tools are now being proposed by NCBI to improve the shortcomings of the classical BlastP . More sensitive search approaches including the search for Hidden Markov Models (HMM) profile, Sequence Similarity Network analysis (SSN), and the use of inferred common ancestor sequences as targets are more appropriate and more useful when it comes to detecting and identifying homologous sequences in any organism. This review highlights a great example of the misleading annotations of proteins in public sequence databases, which is not only limited to MβL fold proteins but applied to many classes of sequences proteins as previously reported, greatly reducing our perception of the multifunctionality of some proteins, due to the unique attributed function by the default protein annotation . In esse


Nitisinone is a medium-sized organic molecule that is used in treating hereditary tyrosinemia type 1 (HT-1). The structurally analogous mesotrione, however, is used as a pesticide/herbicide. What molecular properties are responsible for the similarity/dissimilarity of these molecules is investigated here. The solvent effect reduces the electron affinity to rather negative values and causes the negative electron affinity which manifests itself in a very high positive absolute reduction potential.0, L+, L−) forms of 6 structures. The calculations were conducted in water as a solvent using conductor-like polarizable continuum model (CPCM), nitisinone also in vacuo. The complete vibrational analysis at the true energy minimum allows evaluating the thermodynamic functions with focus to the zero-point energy and overall entropic term. The change of the Gibbs energy on reductions and/or oxidation facilitates evaluating the absolute reduction and absolute oxidation potentials. Also, DLPNO-CCSD(T) method that involves the major part of the correlation energy has been applied to nitisinone and mesotrione and their molecular ions.B3LYP method was utilized for a geometry optimization of nitisinone and mesotrione in their neural and ionized (L It contains 3 rotatable bonds and is hydrophobic, logPow = 0.11 (unbuffered water), acidity constant pKa = 3.12, solubility in water 1500 mg dm−3. It is yellow to tawny solid.Mesotrione (2-(4-(methylsulfonyl)-2-nitrobenzoyl)cycloxexane-1,3-dione, CNitisinone and mesotrione belong to the class of β-triketone herbicides; these are utilized instead of triazine herbicides such as atrazine. Triketones act as inhibitors of the 4-hydroxyphenylpyruvate dioxygenase (HPPD); they, however, increase tyrosine level in plasma. HPPD is an enzyme playing an important role in catabolism of tyrosine.Orfadin) along with restriction of phenylalanine and tyrosine on diet.HPPD, expressed mainly in the liver, converts 4-hydroxyphenyl-pyruvate to homogentisic acid (HGA); it is used in treating hereditary tyrosinemia, type 1 –6. NitisMesotrione is used as a selective herbicide effective mainly to maize. HGA acts as a precursor for synthesis of plastoquinone and α-tocopherol in plants; the inhibition effect to HPPD causes bleaching and death in weed . Mesotriin vacuo and in solvent (water) with focus to redox properties.The present study focuses to the calculations of the electronic structure of nitisinone and mesotrione and to determine molecular descriptors that cause similarity and/or dissimilarity of these species Ab initio calculations have been utilized in order to get electronic structure and molecular descriptors of nitisinone and mesotrione. Two methods were employed as implemented in the ORCA package –11: DFT-εr = 80 for water  = –rGø[J mol−1]/F with the Faraday constant F. The ab initio calculations were conducted also by DLPNO-CCSD(T) method  that account to the major part of the correlation energy. The augmented basis set aug-cc-pVTZ for S-{13,13,13,111/77111/111/11}, F-, O-, N-, C-{88,111/3111/111/11}, and H-{3111/111/11} was applied with 1426 and 1518 basis functions for nitisinone and mesotrione, respectively; the auxiliary basis set was aug-cc-pVTZ/C ; (ii) as a solvent effect, the total energies of the neutral molecule and molecular ions are lowered (items 3 through 5); (iii) upon solvation, the ionization energy is lowered and electron affinity adopts more negative values ; (iv) the chemical hardness is reduced as well (item 8); (v) the polar solvent rises the dipole moment substantially (item 10); (vi) the isotropic value of the quadrupole moment is solvent-independent (item 11); (vii) the isotropic dipole polarizability is raised in water (item 12); (viii) the contributions to thermodynamic functions  are almost insensitive to solvation (items 15–17 and 20–23); (ix) the electronic contributions to thermodynamic functions U, H, and G are influenced by the solvent effect ; and (x) the enthalpy of hydration is ΔhydHø = Hø(L0solv) − Hø(L0vac) =  − 15.7 kcal mol−1 for nitisinone.The set of molecular properties calculated for nitisinone and mesotrione is presented in Table Ei and adiabatic electron affinity Eeg; these allow determining the derived electronic properties such as electronegativity η, and electrophilicity ω. The reaction Gibbs energies serve for determining the absolute oxidation and reduction potentials at standard conditions.Table Ei, Eeg, χ, η, ω) and thermodynamic properties  of nitisinone are essentially the same as for mesotrione. The chemical hardness is a measure of the willingness of the molecule against the electron transfer; for nitisinone and mesotrione, it is η = 39 kcal mol−1 . The electrophilicity index is extremely high: w = 200 kcal mol−1. Consequently, the absolute reduction potential is very high Eredø =  + 3.80 V; for amino acids and small biogenic molecules, it is typically ~  + 1 V when the same methodology (B3LYP) is used. Noticeably, ω is defined on the basis of electronic energies of L0, L+, and L− species and it is common for oxidation and reduction processes. On the contrary, redox potential is derived from the standard reaction Gibbs energies and is different for oxidation and reduction processes.According to Table Gø(Lq) − Eel(Lq) is about 111 and 133 kcal mol−1 which in fact is only 0.01% of the total energy. Therefore, it might be possible to express the redox potential by an approximate formula Er* =  − ΔrEel/F when the Gibbs energy is not available due to a very tedious numerical evaluation of the vibrational frequencies. The ionization energy and/or the electron affinity then serve for the reaction electronic energies.An inspection to Table Q|, isotropic dipole polarizability α, zero-point energy Ezpe, and the total entropic term S·Tø belong to them. The dipole moment reflects a delicate separation of the barycenters of positive–negative charges: p = 6.3 and 8.9 D for nitisinone and mesotrione, respectively.Some of the molecular properties of nitisinone and mesotrione are classified as bulky properties and they increase with increasing molar mass: absolute value of the quadrupole moment |Ei = 198 and 222 kcal mol−1, higher than by M1), and the electron affinity is very negative . High ionization energy is reflected into increased molecular electronegativity χ and increased chemical hardness η. The resulting electrophilicity index is very high  which predetermines high reduction potential.Application of a more exact DLPNO-CCSD(T) method (M2) to above-mentioned neutral and ionized species in geometries optimized by B3LYP method (M1) brings results presented in Table 3 moiety (red colored).The electrostatic potential distributed along the surface of the molecule is taken against a unit charge and is known as the molecular electrostatic potential (MEP) , 24. It −1. The experimental window is 500–4000 cm−1. For nitisinone, the high-frequency domain is limited by medium-intensity bands at I, 2950–3250 cm−1 where a number of low-intensity transitions exist in between. This feature is well reproduced by calculations. There is a broad silent gap between domains I and II. Domain II starts with high-intensity peaks at 1700 cm−1 followed by a huge number of resolved, weakly resolved, or oddly resolved transitions [The calculated IR spectra over the whole wavenumber range are shown in Fig. nsitions . For theII in hexacoordinate form [II to FeIII by strong oxidizing agents (mesotrione and nitisinone) may destroy the functionality of HPPD. However, experiments show that the {HPPD·FeII·NTBC} adduct under aerobic conditions maintains typical electronic transition at 450 nm for a long time, which confirms the presence of FeII [Last but not least, some notes regarding biological activity are given. Both studied compounds, nitisinone and mesotrione, are potent inhibitors of 4-hydroxyphenylpyruvate dioxygenase (HPPD), 40–50 kDa. In active sites, this enzyme contains Feate form , 28. In  of FeII . There ip = 7.3 and 8.9 debye) and bulk properties such as isotropic polarizability (α = 270 and 309 a03), solvated surface and volume, zero-point vibration energy , and the total entropic contribution . The value of the absolute reduction potential is very high (Eredø = 3.80 and 3.82 V) so that these species are effective oxidizing agents. DLPNO-CCSD(T) method, that includes explicitly the correlation energy, applied to nitisinone and mesotrione confirms qualitative predictions of more approximate B3LYP in water: high adiabatic ionization energy, very negative electron affinity , high electronegativity, low chemical hardness, high electrophilicity index , and high dipole moment (p = 7.8 and 9.5 debye).In conclusion, DFT-B3LYP calculations show that two derivatives of β-triketones, nitisinone and mesotrione, possess essentially molecular parameters in water such as energies of HOMO and LUMO, the ionization energy and electron affinity, and derived electronic properties such as molecular electronegativity, chemical hardness, and electrophilicity index. They differ in the dipole moment (high value


We aimed to detect the contribution of cardiac output to  exhibit low peak oxygen uptake (2/∆work rate) and 2 (∆2) were calculated. Normally distributed data were reported as mean ± SD and non-normal data as median [interquartile range].Thirty-five women with FM, aged 23 to 65 years, and 23 healthy controls performed a step incremental cycle ergometer test until volitional fatigue. Alveolar gas exchange and pulmonary ventilation were measured breath-by-breath and adjusted for fat-free body mass (FFM) where appropriate. 2peak was lower in FM patients than in controls . 2 were similar between groups at submaximal work rates, but peak −1, P = 0.005) and C(a-v)O2  were lower in the FM group. No significant group differences emerged in ∆2/∆work rate  or ∆2  slopes. 2 contribute to lower 2peak in FM. The exercise responses were normal and not suggestive of a muscle metabolism pathology.Both https://clinicaltrials.gov/ct2/show/NCT03300635.ClinicalTrials.gov, NCT03300635. Registered 3 October 2017—Retrospectively registered.  The key symptoms of fibromyalgia (FM) include persistent, widespread pain, disturbed sleep, fatigue, and cognitive and mood disturbances . The exa2peak) , and categorical data as count (%), unless otherwise stated. Alpha was set to 0.05. The Weight, BMI, body fat percentage, and waist-to-hip ratio were higher and height lower in the FM group, but there was no difference in FFM between the groups. Patients with FM had higher STAI-trait scores, were less likely to be working, had fewer years of education, and had more comorbidities  than controls. No significant differences were observed in the baseline spirometry values. Background data and spirometry values are shown in Table P = 0.145), SAP , and DAP  were not significantly different between FM and control groups, whereas mean arterial pressure  was higher in the FM group. No significant differences emerged in resting lactate , pyruvate , or L/P ratio  between the groups. Lactate and pyruvate data were missing for four participants in both groups.HR O2, although the between-group differences were small at submaximal work rates. C(a-v)O2 slopes of the two groups were almost identical until peak exercise. SpO2 was within normal range throughout the exercise in both groups, but a group*work rate interaction was noted. Other cardiovascular response slopes are shown in Fig. i, SVR, and SVRi showed no significant group*work rate interactions.Figure P = 0.009, and 93 ± 22 W vs. 118 ± 23 W, P < 0.001) and lower oxygen consumption  than controls. When adjusted for FFM, the difference in oxygen consumption at VT1 was no longer significant , but significance remained at VT2 . 2 at VTs as a percentage of 2peak (VT1% and VT2%) was higher in the FM group . VTs could not be determined for one participant in the FM group, as no clear breakpoints were visible.Participants with FM reached both VT1 and VT2 at lower work rates  fulfilled the RER criteria for maximal effort. Peak HR (HRpeak) and HR as a percentage of predicted heart rate were lower and breathing reserve (BR) higher in the FM group. Peak work rate (Ppeak), 2, 2i, 2, and E were lower in the FM group. PETO2 was lower  and PETCO2 higher  in the FM group. No significant differences were seen in peak E/2, E/2, VD/VT, or SpO2 between FM and control groups. Peak SAP, DAP, and MAP were similar between groups. Oxygen pulse was slightly lower in the FM group . i). Neither SV nor SVi were significantly different between groups at peak exercise. SVR was higher in the FM group, but SVRi failed to reach statistical significance. A significant difference was seen in C(a-v)O2 at peak exercise. Peak exercise results for key parameters are shown in Table me and CTRLme in Table Peak RER and RPE between the groups were comparable. Altogether 25 participants (71%) in the FM group and 21 (91%) in the control group  and L/P ratio , while postexercise pyruvate concentrations were similar  between groups. Data were missing for four participants in both groups.The FM group had lower postexercise lactate concentration  and after exercise , with a median change of 2 . Of the FM patients, 24 (71%) experienced an increase in pain, whereas 8 (24%) reported no change and 2 (6%) a decrease in pain. Data were missing for one participant in the FM group. In the FM group, a negative correlation with baseline pain NRS and 2peak  and PPeak  was observed. Postexercise pain NRS correlated negatively only with 2peak . No significant associations emerged between the change in pain ratings (post–pre) and 2peak  or Ppeak . Neither baseline nor postexercise pain NRS correlated with HRpeak .The FM group reported higher pain NRS at rest  but not with 2peak . Moderate to heavy LTPA correlated with 2peak , HRpeak , and Ppeak  and total LTPA with 2peak . However, no correlation was observed between light LTPA and 2peak , HRpeak  or PPeak . Peak C(a-v)O2 and peak correlated only with moderate to heavy LTPA . When excluding the controls, a significant correlation remained only with total LTPA and 2peak  and moderate to heavy LTPA with 2ipeak . FIQ, PCS, STAI-trait, ACR 2016 WPI, or ACR 2016 SS did nor correlate with 2peak, Ppeak, HRpeak or LTPA in the FM group (data not shown).A positive correlation was observed with n = 7; sleep apnea, n = 2; both, n = 1). Of the seven patients with concurrent FM and asthma, two reached maximal effort. No significant differences emerged in baseline spirometry between asthmatic and non-asthmatic FM patients or between submaximal and maximal groups (data not shown). Group demographics are shown in Table The submaximal group had higher FIQ and STAI-trait scores. More participants in the submaximal group had a pulmonary diagnosis compared with the maximal group. Altogether eight FM patients had pulmonary comorbidities  in FM patients in this study is in concordance with previous studies where a cycle ergometer exercise was used [2peak did not dissipate when adjusted for FFM, demonstrating that lower 2peak in the FM group was not related to body composition. In healthy fit subjects, 2peak is limited primarily by 2 are separate factors in the Fick principle, there is interdependence between the two variables [i 6% lower in the FM group) and peripheral (C(a-v)O2 13% lower in the FM group) mechanisms contributed to lower 2peak in FM, although the difference in peak i was not statistically significant. peak, while SV was similar between groups. Lower HRpeak is a common finding in FM exercise studies [peak respectively with pain ratings, LTPA, and symptom severity in the FM group, but we did not find any correlations. 2peak and Ppeak, however, were negatively associated with baseline pain ratings. C(a-v)O2 is affected by not only oxygen extraction and muscle oxidative capacity but also vascular function and blood flow distribution. Exercise increases bloodflow to the working muscles via peripheral vasodilatation, while sufficient vascular resistance needs to be maintained to ensure adequate MAP [2 could be a consequence of lower mitochondrial oxidative phosphorylation and oxygen demand or lower capillary density in the exercising muscle. Muscle capillary density [2 when analyzing the FM group. Obesity does seem to affect C(a-v)O2 [2 already at submaximal workloads. This study does not provide an explanation for the lower C(a-v)O2, and in clinical settings differentiating mild myopathies from deconditioning may be problematic [2i at VT1, Δ2/ΔP, peak 2, and RER between groups and the lower peak lactate and peak L/P ratio in the FM group, our data do not suggest an impairment in muscle metabolism.The 29% lower was used –43. The subjects . It shouariables . In this studies , 46–48.  studies , 47, 48.uate MAP . MAP and density , mitocho density , and hen density . We did C(a-v)O2 , but if blematic . However2peak. Correlation analysis in our study yielded similar results, suggesting that low moderate to heavy LTPA is a plausible explanation for the lower 2peak in the FM group. Participants in the FM group reported low moderate to heavy LTPA and failed to meet the WHO physical activity recommendation of 150 to 300 min of weekly moderate exercise . A recen2 and work rate. Paradoxical to the fact that exercise training shifts VTs closer to 2peak [peak in relation to predicted maximal HR was lower and peak BR higher in the FM group. Submaximal effort of FM patients has been reported earlier [A few previous studies have reported exercise thresholds (ventilatory or lactate) in FM patients , 43, 46.V˙O2peak , VT1% an earlier , 43, 46.2max) is an important measure of cardiorespiratory fitness representing maximal level of oxidative metabolism. A plateau in 2 occurs near maximal exercise and this is traditionally considered to be the best evidence of achieved 2max [2peak is used instead of 2max. In case a 2 plateau is not attained, secondary criteria are used to determine maximal effort. These criteria most commonly include one or more of the following: HR ≤ 10 - 15 bpm or ≤ 5 - 10% of the age-predicted (220-age) maximum, blood lactate concentration ≥ 8 mM, or RER ≥ 1.00, 1.05, 1.10, or 1.13 [The concept of maximal effort and the issue of possible submaximal effort in the FM group needs to be addressed. Maximal oxygen uptake , the value falls within one SD of the mean of healthy subjects in the study by Beck et al. [2 slope is also well within the normal range of the recently published reference values [The FM patients had a pronounced, albeit not significantly different, circulatory response to increasing oxygen demand, which manifested as steeper ΔHR/Δ to ref. ) and Δ\dent}V˙O2 . Althougk et al. . The ΔHRe values .2peak. In contrast, others have reported an association between cardiorespiratory fitness  and STAI [We noted a possible association between the ability to reach maximal effort and FIQ and STAI-trait Table , althougand STAI  as well and STAI  in FM. Aand STAI . In addi2 between obese and lean subjects is similar when corrected for FFM [2 directly, but rather solved it from the Fick equation, any imprecision in measuring cardiac output would additionally impact our C(a-v)O2 results. The study population consisted of only women, and our results cannot be extrapolated to male FM patients.Although the sample size was adequate for the primary outcomes, the subgroups in our secondary analysis were small, diminishing statistical reliability. The patient and control groups were not entirely homogeneous regarding their anthropometrics, educational and employment status. This reflects real-life differences between patients with FM and their same-aged peers. Although obesity has a multitude of systemic effects, adipose tissue does not affect oxygen uptake during exercise, and  for FFM , 65. Gat for FFM , 66, 67. for FFM , 68, 69, for FFM  and chro for FFM . MoreoveThe main strength of this study lies in the simultaneous recording of ventilatory gas exchange and ICG data. To the best of our knowledge, exercise responses in patients with FM have not been studied this intensively before.2peak is deconditioning and less moderate to heavy LTPA.Patients with FM display poor cardiorespiratory fitness and both cardiac output and arteriovenous oxygen difference were lower compared with healthy controls. Abnormal muscle metabolism seems unlikely, whereas a possible explanation for the observed lower


The evidence on the course of the intelligence quotient (IQ) at the long term in individuals with schizophrenia spectrums disorders is inconclusive.We aimed to analyse whether IQ improves, declines, or remains stable over 10 years in a sample of patients with First Episode Psychosis (FEP) and healthy controls (HCs).The FEP patients participated in a Program of First Episode Psychosis in Spain called PAFIP. At baseline, FEP patients provided demographic and clinical data, and completed a neuropsychological assessment that included an estimation of premorbid IQ trough the WAIS vocabulary subtest. At 10-year follow-up, the participants were invited to complete the same evaluation and 10-year IQ was estimated. The group of HCs underwent the same neuropsychological battery at both moments. Cluster analysis was performed separately in the FEP patients and the HCs to determine their profiles of intellectual change.FEP patients (n=137) were grouped into five clusters : “Improved low IQ” (9.49% of patients), “Improved average IQ” (14.6%), “Preserved low IQ” (17.52%), “Preserved average IQ” (43.06%), and “Preserved high IQ” (15.33%). Ninety HCs were grouped into three clusters: “Preserved low IQ” (32.22% of the HC), “Preserved average IQ” (44.44%), and “Preserved high IQ” (23.33%). Demographic data of FEP patients are presented in Table 1.The FEP patients showed intellectual improvement or stability, but no decline post-onset of psychosis. However, their profiles of intellectual change are more heterogeneous than that of HCs over 10 years. Particularly, there is a subgroup of FEP patients with a significant potential for long-term cognitive enhancement.None Declared


The “Med-fusion” DPP had the lowest protein-to-fat ratio (p < 0.001). Infants of women following the “Dairy-focused” DPP had the highest birth height centiles (p = 0.007) and the lowest ponderal index (p = 0.003). The NMR-metabolomics approach was implemented on a subset of women that provided amniotic fluid (AF) specimens (n = 62) to elucidate distinct metabolic signatures associated with DPPs. PCA and OPLS-DA models verified the adherence to three DPPs, revealing that the levels of several amino acids (AAs) were the highest in “Dairy-focused”, reflecting its protein-rich nature. The “Traditional-inspired” DPP showed decreased AAs and glucose levels. This knowledge may contribute to optimizing maternal dietary recommendations. Further research is needed to validate these findings and better understand the relationships between maternal diet, AF metabolic signature, and neonatal anthropometrics.This study aimed to characterize dietary protein patterns (DPPs) in a sample pool of 298 well-nourished pregnant women and explore potential associations between DPPs and neonatal anthropometrics. Maternal dietary data were collected using a validated food frequency questionnaire. Neonatal anthropometrics were abstracted from health booklets. A hierarchical cluster analysis identified three DPPs: “Dairy-focused”, “Med-fusion”, and “Traditional-inspired”. The “Dairy-focused” DPP exhibited the highest protein intake ( It is well-established that maternal nutrition during pregnancy is a major environmental stimulus that can alter fetal morphology and physiology, resulting in different phenotypes that affect offspring survival and long-term health ,2,3. In A large body of epidemiological studies has indicated a positive association between maternal protein intake and neonatal anthropometric characteristics ,10,11,12New potentials in nutritional science are offered with the development of highly sensitive analytical platforms and (bio)informatics. Specifically, the implementation of metabolomics in various biological matrixes is an efficient tool for exploring the effects of perinatal nutrition on pregnancy evolution and outcome ,28,29,30As such, the primary objective of the current study was to explore potential associations between maternal DPPs in the second trimester of pregnancy and neonatal anthropometrics. The secondary objective pertained to the implementation of NMR metabolomics in a subset of participants that provided AF specimens to comprehensively evaluate metabolic signatures related to maternal DPPs.A. Materials and Methods Regarding the Primary ObjectiveThree hundred twenty-seven pregnant women who attended the 1st Department of Obstetrics and Gynecology, Papageorgiou General Hospital in Thessaloniki (Greece), for a scheduled prenatal visit were initially invited to participate in the present prospective study. To be eligible for participating in the survey, women had to meet the following criteria: a. be more than 18 years of age, b. be familiar with the Greek language, c. have a singleton pregnancy, d. be in the second trimester of pregnancy at the time of enrollment, and e. be, apparently, healthy . The study design, the flow of participants, and the key processes applied in the current study are schematically depicted in Initially, women were informed about the objectives of the current study. Those who agreed to participate in the research study gave their signed consent and provided information regarding sociodemographic characteristics. Maternal anthropometric measurements were also taken, and women completed a structured interview concerning maternal dietary habits. Further details as far as the data collection process is concerned are presented in The procedures followed were in accordance with ethical approval attained from the Bioethics Committee of the Medical School, Aristotle University, Thessaloniki, Greece (A19479—26/2/08), and in compliance with the declaration of Helsinki.2). Participants were categorized into BMI groups based on classification criteria provided by the World Health Organization (WHO)  [All women provided information on age, occupation, education, pre-pregnancy anthropometrics (height and weight), level of physical activity (PA), and smoking history . Educati0 kg/m2) . Smokers0 kg/m2) .Maternal dietary intake in the second trimester was assessed using a Mediterranean-oriented semi-quantitative food frequency questionnaire (FFQ), previously validated among 179 pregnant women in Greece . The FFQInformation regarding childbirth , neonate gender, and anthropometric characteristics (birth weight and height) were retrieved from the newborn’s health booklet during a structured telephone interview with a well-trained interviewer. In Greece, newborn health booklets are filled out by medical staff immediately after delivery and are given to all mothers upon leaving the hospital. The ponderal index was calculated by dividing the weight (g) by the third power of the height (cm) and multiplying it by 100 . Birth wn = 327) for one of the following reasons: a. were diagnosed with pregnancy complications, such as gestational diabetes mellitus, pregnancy-induced hypertension, preeclampsia, etc. (n = 4), b. had terminated pregnancy (n = 2), c. had preterm delivery (n = 5), d. had born infants with structural malformations, chromosomal abnormalities, and/or congenital conditions that could affect fetal growth or development (n = 7), or e. could not be located or dropped out (n = 3).As depicted inA Microsoft Excel database was used to transform information collected regarding dietary data into daily intakes . During A hierarchical cluster analysis (HCA) ,38 was aTo calculate protein intake (g/day) for each of the 298 participants, the amount (g) of each food consumed daily was multiplied by the protein content (g) of this specific food.To convert these intakes into the percentage (%) of energy derived from protein, the formula given below was applied:To facilitate the interpretation of HCA, foods were classified, according to their protein content as well as practices/preferences reflecting dietary habits, into 19 predefined and mutually exclusive food groups  40,41,4,43.10 (X + 1) transformed to remove the potential extraneous effect of variables with the largest variances as well as to achieve homogeneity of variance [The percentages of energy derived from protein for the 19 food groups were logvariance .To create the input variables, the procedure described below was followed:These 19 log-transformed values for the 298 participants were used as input variables in the HCA.a = 0.05 (p = 0.05)), (b) the size  and the interpretation of each DPP, and (c) the tree diagram resulting from the Ward method of HCA [The three-cluster solution was selected as the best-case scenario.The number of DPPs selected, after performing several runs of cluster formation, was based on the following criteria: (a) the number of input variables with statistically significant differences in means .All statistical analyses were performed with IBM SPSS Statistics software, Version 28.0. For all hypothesis testing procedures, the significance level was predetermined at n) and the corresponding percentages (%) for categorical data. Food group consumption and nutrient intake data were presented as mean ± SD.Demographic/anthropometric and selected lifestyle characteristics of women were presented as the mean ± standard deviation (SD) for quantitative data and as the number of subjects ; cluster membership was treated as the independent variable, while the consumption of food groups as the dependent variable. The R2 value indicates the percentage (%) of the variance in the investigated food group consumption explained by the differences among DPPs. In the methodological frame of ANOVA, R2 is computationally and conceptually equivalent to the “Eta-squared” (η2) index, a measure of the independent variable’s effect size  [η2 = R2 = , where SS is the corresponding sum of squares [The contribution of each of the 19 food groups in the cluster construction was evaluated by investigating the magnitude and the statistical significance level of the corresponding coefficients of determination, bership) . Eta-squ squares .χ2) for categorical variables. The significance level  for χ2 was calculated with the Monte Carlo simulation method using 10,000 random samples. This methodological approach leads to safe inductive conclusions even in cases where the methodological assumptions of χ2 are not fully met [The homogeneity of variance among the three DPPs was examined using Levene’s test. To compare differences across DPPs in food groups, nutrient intake, and maternal and birth outcome data, the AΝOVA method followed by the Tukey’s or the Games–Howell post hoc test (when the homogeneity of variance assumption was violated) was used. DPPs were compared using the chi-squared test  was established using the last menstruation date confirmed with ultrasound scanning. After the biochemical analysis for cytogenetic-based diagnostics was realized, aliquots from the residual AF were used for NMR metabolomics .The analysis of AF samples was carried out using nuclear magnetic resonance (NMR) spectroscopy  followin2O and 150 μL phosphate buffer in D2O were added to lyophilized AF samples. After centrifugation , 50 μL of sodium maleate was added as an internal standard to 500 μL of the supernatant, and the sample was transferred to 5 mm NMR tubes.For the AF sample preparation, 400 μL DSodium maleate was chosen as the reference standard since it is suitable for the CPMG pulse sequence and provides a distinct peak in the 1 H NMR spectrum. The samples were thawed at room temperature for 60 min before performing NMR experiments.All NMR spectra were acquired using a Varian-600MHz NMR spectrometer equipped with a triple resonance probe {HCN} at 25 °C. The Carr–Purcell–Meiboom–Gill (CPMG) pulse sequence was applied to AF samples with 128 transients collected with 64 K data points. The relaxation delay was set to 6 s. The receiver gain was kept constant for all acquisitions. Proton spectra were referenced at the resonance peak of sodium maleate (5.95 ppm).1H-NMR spectra were preprocessed with the MestreNova (v.10.1) software . Manual phase correction, automatic baseline correction, and sinc apodization were applied to improve spectra resolution. Total area normalization and binning of 0.0001 ppm were selected. A superimposed spectrum was constructed, and the peaks were manually aligned. The water D2O region (4.68–5.00 ppm) was excluded.All Metaboneer, an in-house, fully automated metabolite identification platform , facilitA series of 2D experiments, i.e., gCOSY, zTOCSY, gHMBCad, and gHSQCad experiments, were recorded at 25 °C and permitted the unambiguous assignment of metabolites; their acquisition parameters are described in Postprocessing of Spectral Data: SIMCA-P  was facilitated. The spectral data were mean-centered Pareto-scaled (Par), and the principal component analysis (PCA) and orthogonal partial least squared discriminant analysis (OPLS-DA) models were extracted at a confidence level of 95%. The mathematical background and applications of these methods have been extensively discussed [iscussed .Identification of Important Features: S-line plots were used to pinpoint the metabolites that contributed to the samples’ discrimination.Model Validation: The quality of models (PCA/OPLS-DA) was described using the goodness-of-fit R2 (0 ≤ R2 ≤ 1) and the predictive ability Q2 (0 ≤ Q2 ≤1) values. The R2 explained the variation, thus constituting a quantitative measure of how well the data in the training set were mathematically reproduced. The overall predictive ability of the model was assessed using the cumulative Q2, representing the fraction of the variation in Y that could be predicted using the model, which was extracted according to the internal cross-validation default method of the software SIMCA-P. Q2 is considered a de facto default diagnostic parameter for validating OPLS-DA models in metabolomics. In particular, the difference between the goodness of fit and the predictive ability always remained lower than 0.3 (R2X(cum) − Q2 (cum) < 0.3), and the goodness of fit never equaled one (R2X(cum) ≠ 1). The extracted models abided by these rules; therefore, their robustness and predictive response were enhanced and over-fitting was effaced.p-value < 0.05. Permutation tests were used (999 permutations) to evaluate whether the specific classification of two classes in a model was significantly better than any other models obtained by randomly permuting the original group’s attribution. An additional measure of PLS-DA model validity included the extraction of receiver-operating characteristic (ROC) curves to assess the ability of the PLS latent variable T pred to correctly classify the test set. A marker explained a low, fair, and superior diagnostic accuracy when the area under the ROC (AUROC) curve reached values of 0.5 < AUC < 0.7, 0.7 < AUC < 0.9, and AUC > 0.9, respectively. The area under the ROC (AUROC) was calculated. A perfect discrimination corresponded to an AUROC equal to 1 [Regression models were validated using cross-validation analysis of variance (CV-ANOVA) with a ual to 1 ,52.Metabolic Pathways: The online platform MetaboAnalyst RRID:SCR_015539  [As already mentioned, from the 298 women who finally participated in the study, 62 women also provided AF samples for NMR metabolomics analysis . For thin = 74), “Med-fusion” (n = 104), and “Traditional-inspired” (n = 120)) (n = 9), “Med-fusion” (n = 23), and “Traditional-inspired” (n = 30)) were compared with the corresponding 95% bootstrap CIs for each of the three DPPs (“Dairy-focused” (n = 74), “Med-fusion” (n = 104) and “Traditional-inspired” (n = 120)).In more detail, initially, for each of the three extracted DPPs from the whole sample (“Dairy-focused” ( = 120)) , 95% booA. Results Regarding the Primary Objective2, and 65.1% were of normal weight. Less than 10% of the study population was obese.Selected characteristics of the 298 pregnant women participating in the current prospective study are summarized inn = 74), “Med-fusion” (n = 104), and “Traditional-inspired” (n = 120). Selected sociodemographic and anthropometric characteristics of the women in each DPP are listed in After applying HCA to the log-transformed percentage of energy derived from protein intake, three DPPs were identified. The patterns were named as follows: “Dairy-focused”  of the energy derived from protein.p ≤ 0.05) were observed. The most profound differences . The “Dairy-focused” DPP presented the highest intake of “low-fat dairy products” (2.92±1.41%), whereas the “Med-fusion” DPP consumed more “full-fat dairy products” (1.85±1.37%) compared with the two other DPPs. The “Traditional-inspired” DPP was characterized by an increased intake of “whole grain cereals” (1.50±0.58%) and intermediate consumption of both “low-fat dairy products” and “full-fat dairy products” .As listed inferences A–C, as sNotably, women in the “Dairy-focused” DPP also preferred consuming other dairy products, such as yellow cheese , while in the case of the “Med-fusion” DPP, the elevated intake of “Refined cereals” was accompanied with an increased preference for all starchy foods . Women in the “Traditional-inspired” DPP favored more nutritious plant-based products, such as legumes and nuts.p = 0.004) and as a percentage of total energy intake (%E) (p < 0.001) compared with the “Med-fusion” and “Traditional-inspired” DPPs. Differences were also reported regarding the quality of protein consumed. Individuals following the “Dairy-focused” DPP presented the highest animal protein intake (mean different: ~8 g), while those in the “Traditional-inspired” DPP consumed much more plant protein (6.12 ± 0.95%E) compared with those following the “Dairy-focused” and the “Med-fusion” DPP .Differences in food group consumption between DPPs were mirrored in the nutritional profile as well as the indices of dietary quality . IndividBeyond the variations in protein, the different dietary behaviors were also reflected in the “whole protein package” . ParticiIn No statistically significant differences were noted between the DPPs regarding gestational age or neonate gender.p = 0.601), and height (p = 0.279) (p = 0.007). This statistically significant difference was also accompanied by a slightly lower ponderal index.Women following the “Dairy-focused” DPP gave birth to infants with slightly lower ponderal index (2.39 ± 0.26 g/cm3) than those following the “Med-fusion” (2.49 ± 0.25 g/cm3) and “Traditional-inspired” DPPs (2.52 ± 0.23 g/cm3) (p = 0.003).No statistically significant differences were found among the three DPPs for birth weight (= 0.279) .HoweverB. Results Regarding the Secondary Objectiven = 62) to the full dimensions of the sample (n = 298) was to compare the dietary databetween the initial DPPs, derived from the whole sample , in response to a dietary pattern, were aminoacyl-tRNA biosynthesis, valine, leucine and isoleucine biosynthesis, histidine metabolism, pantothenate and CoA biosynthesis, beta-alanine metabolism, alanine, aspartate and glutamate metabolism and valine, leucine and isoleucine degradation. Of these, the “histidine metabolism” and the “alanine, aspartate, and glutamate metabolism” pathways had the greatest impact.Our results revealed that the primary disturbed statistically significant pathways , the interpretation of their dominating presence in the “Dairy-focused” AF specimens seems to be supported by the existing literature ,74,75,76The assumption that the maternal diet contributes to the pool of free AAs in AF could also be sound in the case of histidine. The human organism, during intrauterine life, has not yet developed the ability to produce histidine . Indeed,Providing explanations regarding the presence of several dispensable AAs  in elevated levels in the “Dairy-focused” DPP compared with the other two DPPs is rather a laborious process. So far, several studies conducted among pregnant  and non-Glucose also appears to be a metabolite associated with the dietary habits of the population under study. A plausible explanation for this observation may be also associated with the specific protein sources consumed. As has already been discussed, women in the “Traditional-inspired” DPP compared with the other DPPs exhibited a higher preference for plant protein sources, such as whole grain cereals, legumes, and nuts; a dietary choice that is directly reflected in the intake of dietary fibers and the quotient of carbohydrate-to- fiber. As such, it can be postulated that women in the “Traditional-inspired” DPP may exhibit a more gradual rise in postprandial glucose levels than the other DPPs, resulting in differential glucose fluxes from the mother to the developing fetus through the placenta. Similar findings have also been reported in previous studies conducted both in humans  and animThe findings of the current study should be considered in the context of potential strengths and limitations. Notably, the implementation of a protein-centric approach to identify maternal dietary patterns is an innovative aspect of this work. This methodological framework enables a more comprehensive understanding of the potential associations between maternal protein intake during the second trimester of pregnancy and fetal growth and development. A noteworthy advantage of our study is the use of a validated FFQ for dietary data collection, which was indeed completed through personal interviews by a registered dietician or a well-trained interviewer. This approach ensured to a great extent the accuracy and reliability of the obtained information . Moreoven = 62) revealed unique metabolic signatures associated with each DPP, indicating that not only proteins but also the “whole protein package” might have a differential effect on metabolic processes. Moreover, these findings highlight the importance of investigating DPPs during pregnancy, as they may be related to the risk of non-communicable diseases in later life. However, further research with larger sample sizes and comprehensive metabolomic analyses is urgently warranted to validate these findings and better understand the associations between maternal DPPs, neonatal anthropometrics, and AF metabolic signatures. This knowledge may also contribute to optimizing maternal dietary recommendations regarding optimal protein intake and the proportions of proteins derived from different sources.In conclusion, the findings of the current prospective study suggest that maternal protein intake, when investigated from the whole-diet approach, may influence neonatal size, as depicted in birth height centiles and the ponderal index. The comparative analysis of AF specimens (


Visual perspective taking (VPT) represents how the world appears from another person’s position. The age, group status and emotional displays of the other person have been shown to affect task performance, but tasks often confound social and spatial outcome measures by embedding perspective taking in explicitly social contexts or theory-of-mind reasoning. Furthermore, while previous research has suggested that visual perspective taking may be impacted by avatar characteristics, it is unknown whether this is driven by general group processing or a specific deficit in mentalizing about outgroups, for example, children. Therefore, using a minimally social task , we examined whether avatar age and avatar gender affect performance on simpler (low angular disparity) and more effortful, embodied (high angular disparity) perspective judgments. Ninety-two participants represented the visuospatial perspectives of a boy, girl, man, or woman who were presented at various angular disparities. A target object was placed in front of the avatar and participants responded to the orientation of the object from the avatar’s position. The findings suggest that social features of visuospatial perspective taking (VSPT) are processed separately from the fundamental spatial computations. Further, Level-2 VSPT appears to be affected by general group categorization  rather than a deficit in mentalizing about a specific outgroup .The online version contains supplementary material available at 10.3758/s13423-023-02249-7. Specifically, it will illuminate social and perceptual processing integration, explore the limits of avatar-characteristic influence in L2-VSPT, and determine whether general group processing or age-dependent mentalizing explains social processing effects in L2-VSPT. Gender and age were manipulated for three reasons. First, gender and age are easily accessible physical characteristics , if general group processes exert influence over visuospatial processing, similarity may be facilitative , if general group processes exert influence over visuospatial processing, significantly different response times are hypothesized between same-gender adult avatars and others. Replicating previous findings  were recruited online via opportunity sampling (n = 39) and Prolific Academic (n = 60) and received £3.00 remuneration for ~20 min of their time. The experiment was granted ethical approval by the Departmental Research Ethics Committee. Participants that did not finish the task (n = 2), did not complete demographic information (n = 1), did not meet eligibility criteria (n = 1), identified as a gender other than male or female (n = 1), or had < 66% accuracy (n = 2) were excluded from further analysis. This left a total of 92 participants  whose data was able to be included in the analysis.For linear mixed effects models, to discover an effect analogous to Pavlovia.org  sitting in front of, and looking at, a dog. Experimental stimuli are based on Surtees et al.  but withThe study used a 2 × 2 × 3 repeated-measures design with avatar age (adult/child), avatar gender (same/other), and angular disparity 45°/90°/135°) as factors. To code the avatar gender, participants who identified themselves as female had “woman” and “girl” stimuli recoded to “same-gender” and “man” and “boy” stimuli recoded to “other-gender,” and vice versa. To provide the greatest opportunity for social processing, the decision was made to position the avatar centrally for maximum saliency  and, after agreeing to participate, they were redirected to the task on To facilitate good quality data, participants recruited from Prolific Academic had declared themselves as fluent speakers of English, completed at least ten prior studies, and had 100% approval ratings. The number of trials per condition was maximized to account for the expected variability from collecting data online whilst keeping block duration (~3 min) and overall participation time (~20 min) brief to facilitate attention  for R  and a significant fixed effect of 135° . Further analysis confirmed significantly less accuracy at 135°  compared to 90°, OR = 5.24, p < .001. No other fixed effects or interactions were significant  and 135° . No fixed effects of age or gender reached significance (both ps > .07). However, there was a significant interaction between age and gender, β = -0.02, t = -2.06, p = .04. Figure Analyses found significant fixed effects of 90° ((how the world appears). Accordingly, this study is the first to use a minimally social task to explore the effects of agent characteristics in L2-VSPT. Results suggest that implicit processing of avatar age modulates response times only for same-gender avatars. Explanations address how social processing may be incorporated into L2-VSPT, provide support for the general group processing hypothesis rather than a diminished salience of children’s minds, and help determine the limits of avatar-characteristic effects.Previous research suggests that social features affect VSPT task performance, but confounded measures mean it is difficult to disentangle whether social cues impact fundamental perspective computation or the communicative and social elements of tasks. Furthermore, despite growing emphasis on agent characteristics in L1-VPT (visibility judgments), little attention has been given to L2-VSPT The significant fixed effect of angle found for accuracy and response times replicates a considerable amount of previous work that suggests that cognitive effort increases with increasing angular disparity and, thus, accuracy decreases, and response times increase


Pharmaceutical agonists of peroxisome proliferator-activated receptors (PPARs) are widely used in the management of type 2 diabetes, chiefly as lipid-lowering agents and oral hypoglycaemic agents. Although most of the focus has been placed on their cardiovascular effects, both positive and negative, these agents also have significant renoprotective actions in the diabetic kidney. Over and above action on metabolic control and effects on blood pressure, PPAR agonists also appear to have independent effects on a number of critical pathways that are implicated in the development and progression of diabetic kidney disease, including oxidative stress, inflammation, hypertrophy, and podocyte function. This review will examine these direct and indirect actions of PPAR agonists in the diabetic kidney and explore recent findings of clinical trials of PPAR agonists in patients with diabetes. Over the last decade, PPARγ agonists, known as thiazolidinediones (TZDs) or glitazones, have also come into clinical use as oral hypoglycaemic agents. Selective agonists of a third isoform of PPAR, PPAR β/δ are also under clinical development for treatment of the metabolic syndrome . Im. Imα agofindings –31. By cype mice . P < 0.05) . In. Inα agoofibrate . These fofibrate . Anotherofibrate , 57, whidiabetes . Recent  actions .α agonists exert a range of anti-inflammatory actions which may be beneficial in diabetic kidney disease. For example, treatment with fibrates is able to suppress circulating levels of cytokines in patients with diabetes . Th. Thγ agoty) rats –31. In oty) rats . Even inving GFR .α agonists, approximately 10–20%, and appeared to be independent of improved glycaemic control. However, treatment with the PPARγ agonist, pioglitazone was associated with greater decline in estimated GFR than with placebo [A large number of clinical trials have demonstrated that TZDs can also reduce albuminuria patients with type 2 diabetes. A recent meta-analysis including 15 of these randomised controlled studies (5 with rosiglitazone and 10 with pioglitazone) suggested that TZD treatment, was associated with a significant fall in urinary albumin excretion . The mag placebo .Further evidence for a role of PPAR in diabetic kidney disease also comes from polymorphism studies in patients with diabetes and nephropathy. For example, the Pro12Ala polymorphism has been linked to a higher incidence of nephropathy in patients with type 2 diabetes , 82. Theγ activation results in increased sensitivity to the metabolic actions of insulin, partly by reversing lipotoxicity-induced insulin resistance. In addition, TZDs have been shown to “rejuvenate” pancreatic β-cells, reducing β-cell apoptosis, and increasing β-cell proliferation, thus maintaining β-cell mass and function [γ activation may indirectly improve renal function. In addition, there is some evidence that insulin resistance plays a more direct role in the pathogenesis of diabetic renal disease. In particular, pathway-selective insulin resistance appears to be associated with activation of pathogenic pathways due to hyperinsulinism [β expression in the kidney are partly attenuated by pioglitazone, without affecting glycemic control [PPARfunction . Resistasulinism . Indeed,sulinism . In expe control .γ agonists also appear to have direct effects on a number of critical pathways that are implicated in the development and progression of diabetic kidney disease, over and above action on metabolic control. None of these actions occurs in isolation, but they appear synergistically intertwined, consistent with the key role of the PPARγ signalling pathway as a metabolic and cellular regulator.PPARγ activation by TZDs commonly reduces blood pressure and prevents the development of hypertension [γ gene are associated with hypertension in humans [Agonists may also directly impact on blood pressure –89, one rtension , 91. In n humans , 93. It n humans , 90. Howγ agonist, rosiglitazone [γ agonist rosiglitazone or PPARγ overexpression is also able to protect against podocyte injury inducted by aldosterone [γ activation is also able to attenuate AGE-induced ROS generation and decrease RAGE expression [We have previously shown that NADPH-dependent superoxide production is increased in the vasculature of diabetic mice and that this may be attenuated following treatment with the PPARlitazone , 95. Thilitazone , and imposterone , potentiosterone . PPARγ apression .γ agonists, rosiglitazone, is able to reduce renal hypertrophy associated with STZ diabetes [γ is able to directly modify cell cycle signaling [γ agonists. PPARAs noted above, diabetes is characterized by maladaptive renal hypertrophy. We have previously shown that treatment of diabetic mice with the PPARdiabetes  and unindiabetes . It has ignaling . Certainignaling . Other eκB activation, and macrophage infiltration in the kidney [Inflammation also plays a significant role in the development and progression of diabetic kidney disease. Treatment with TZDs is able to suppress circulating levels of cytokines in patients with diabetes , 61. It e kidney .in vitro [α agonists are also able to increase gene expression of the key slit-pore protein, nephrin, in diabetic nephropathy, potentially contributing to their antiproteinuric actions [γ agonists also affect nephrin gene transcription [γ agonist, pioglitazone, had an antiproteinuric effect, possibly via transcriptional regulation of nephrin [The glomerular podocyte plays a key role in the development and progression of albuminuria and glomerulosclerosis associated with diabetes –69. Suchin vitro  and in vin vitro , 72–74.  actions . PPARγ acription . Similarβ and CTGF, as well as metalloprotease activity. Certainly, treatment with PPARγ agonists is able to reduce glomeruloscleoriss and tubulointerstitial fibrosis in diabetic mice  cholesterol [Some studies have suggested that there is an association between the metabolic phenotype and polymorphisms of the PPARδ gene . In humalesterol . In theolesterol –41. Howeα may be partly responsible for the renoprotective actions of these agents. Activation of the PPARδ receptor is thought to alter the circulating lipid profile by enhancement of fat oxidation in skeletal muscle. PPARδ knockout mice are also insulin resistant/glucose intolerant, although this may reflect lipotoxicity rather than a direct effect on insulin sensitivity. PPARδ agonists may also have direct effects on a number of critical pathways that are implicated in the development and progression of diabetic kidney disease, over and above action on metabolic control.As noted above, dyslipidaemia is a major reversible risk factor for diabetic CKD –41. The δ is involved in the regulation of apoptosis in response to renal injury [δ increases susceptibility to ischemic injury in the kidney, while selective agonists of PPARδ protect against ischemic acute renal failure [δ is increased in the diabetic kidney [Some data suggest that PPARl injury . Deficie failure . Certain failure , implyinc kidney .δ receptor appears to have anti-inflammatory actions in the kidney [δ binds to the anti-inflammatory transcriptional repressor Bcl-6, inhibiting its suppressive actions on inflammation [δ agonist GW0742 reduces macrophage infiltration in the diabetic kidney [δ is not essential for the anti-inflammatory effect of some of the so-called selective PPARδ receptor ligands, suggesting that other pathways may also be involved [Activation of the PPARe kidney . It is kammation . Certainc kidney . Whetherinvolved .δ receptor in diabetic mice is reduced following treatment with the PPARδ agonist, L-165041 [in vitro [The AGE/RAGE pathways has been strongly implicated in the pathogenesis of diabetic kidney disease. Activation of the PPARL-165041 . Certainin vitro , partly PPAR agonists have clear and reproducible renoprotective effects, impacting on a range of pathways implicated with the development and progression of diabetic kidney disease, including oxidative stress. For the TZDs, the cardiovascular toxicity, fluid retention, and risk of bladder cancer have now overshadowed these important renoprotective actions. However, the new recognition of fibrates as agents that prevent microvascular complications of diabetes will hopefully leads to renewed interest in the role of PPARs in the kidney.α/γ agonists is also salutary. Although many times more effective activators of PPAR receptors, these agents appeared no more effective than currently available agonists, while having a poorer side effect profile. It may be that PPAR agonists also have activities through PPAR-independent pathways, possibly via the transrepression of pathogenic genes, such as activator protein-1 (AP-1) and nuclear factor-κB (NF-κB) [The actions of the glitazars, highly potent combined PPAR  (NF-κB) . Such ac (NF-κB) . Greater


A comparison among two forms of half-diallel analysis was made. The different half-diallel techniques used were Griffing's model I, method 2 and 4. These methods of diallel analysis were found to be interrelated. However, as Griffing's model I, method 4 partitioned heterosis into different components as well as gave information about combining ability and this method had certainly some advantages over the other. The results further indicated using parental generations in the second Griffing method may cause biased estimate of the GCA and SCA variances. Thus, using the fourth Griffing method is more suitable than the other methods in providing time, cost, and facilities, and it is recommended as an applicable method. Estimates of combining ability are useful in determining the breeding value of cucumber lines by suggesting the appropriate use in a breeding program. In studying combining ability, the most commonly utilized experimental approach is the diallel design. In the diallel analysis, Sprague and Tatum  introduci, j). Similarly, specific combining ability represents the dominance deviation value in the simplest case ignoring epistatic deviation; see Kempthorne [Griffing  defines mpthorne  and Mayompthorne  for detap inbred lines. Gupta and Kageyama [p, is large, or reciprocal crosses are similar to direct crosses it becomes impractical to carry out an experiment using a complete diallel cross design. In such situations, we use partial diallel cross designs where a subset of crosses are used. Although efficient designing of partial diallel crosses has been studied by several authors [Complete diallel cross designs involve equal numbers of occurrences of each of the distinct crosses among Kageyama , Dey andKageyama , and DasKageyama  investig authors –11, no f authors  and Das  authors . Sometim authors  and Muke authors  provide In the present paper, a comparative view of Griffing's model I, method 2 and 4 has been presented and discussed in light of their practical significance.(Cucumis sativus L.) were produced. The varieties used were (1) “BH-502”, (2) “BH-504”, (3) “BH-604”, (4) “BH-605”, (5) “08wvc c-115”, (6) “08wvc c-118.” These crosses, along with their parents, were evaluated in a randomized block design with three replications. The following characteristics were recorded: early, unmarketable, marketable, and total yield; simple weight index (SWI). Simple weight index was calculated following Wehner and Cramer [To start with, 6 × 6 half diallel crosses of cucumber Griffing's model IXij = u + gi + gj + sij + (1/b)∑keijk, (i) Method 2: Xij = u + gi + gj + sij + (1/b)∑keijk (ii) Method 4: i = j = 1 … p; k = 1 … b), where u = the population mean; gi = the general combining ability effect of the ith parent; gj = the general combining ability effect of the jth parent; Sij = the specific combining ability effect of the cross between ith and jth parents such that slj = sji; eijk = the environmental effect associated with ijk th observation. . In fact, this result indicated that method 4 is more suitable than method 2. Some authors believe that when the differences between hybrids and parents are significant, method 4 without parents entering in estimations is better that method 2 . They in