Cystinosis is a rare inheritable lysosomal storage disorder characterized by cystine accumulation throughout the body, chronic kidney disease necessitating renal replacement therapy mostly during adolescence, and multiple extra-renal complications. The majority of male cystinosis patients are infertile due to azoospermia, in contrast to female patients who are fertile. Over recent decades, the fertility status of male patients has evolved from a primary hypogonadism in the era before the systematic treatment with cysteamine to azoospermia in the majority of cysteamine-treated infantile cystinosis patients. In this review, we provide a state-of-the-art overview on the available clinical, histopathological, animal, and in vitro data. We summarize current insights on both cystinosis males and females, and their clinical implications including the potential effect of cysteamine on fertility. In addition, we identify the remaining challenges and areas for future research. CTNS gene. This gene encodes the protein cystinosin, which is a lysosomal membrane protein responsible for transporting cystine, produced by the degradation of proteins in lysosomes, from the lysosome into the cytosol. Hence, mutations in CTNS cause the intralysosomal accumulation of cystine, leading to various effects in the body [Cystinosis is a rare autosomal metabolic disorder caused by bi-allelic mutations in the the body . The agethe body ,3. In ththe body ,3. One othe body .For clinical fertility management purposes, azoospermia is classified into obstructive (OA) and nonobstructive azoospermia (NOA), based on clinical sexual characteristics (testicular volume) and sex hormone levels . In OA, Due to the advances in the treatment of cystinosis patients over recent decades, their life expectancy has substantially increased, making fertility a new and important issue for both patients and their treating physicians . In thisWomen with cystinosis have been reported to suffer from delayed puberty, with a menarche around the age of 15–19 years, while a stable menstrual cycle is reached at least two years after menarche . In accoUnlike men suffering from cystinosis, fertility seems to be unaffected in female cystinosis patients. Several reports have described successful pregnancies in women with cystinosis ,11,12,13Another important issue in female cystinosis patients who want children is the teratogenicity of cysteamine. Since this is a lifesaving treatment, the timing of its discontinuation should be well thought out. The current advice is to stop cysteamine as soon as a pregnancy test is found to be positive . HoweverCTNS gene, the chances that the child will develop cystinosis are minimal. Importantly, some mutations can be difficult to detect (such as deep intronic mutations), so couples should be counseled that there is still a very small likelihood that the child will have cystinosis if such a mutation remains undetected in the patient’s partner.In general, it is advised for women with cystinosis who want to become pregnant to seek medical advice in advance, preferably in a clinic with expertise in pregnancies in patients with chronic kidney disease or kidney transplantation. By doing so, teratogenic medications can be switched before conception and a plan can be made when cysteamine has to be stopped. Since cystinosis is an autosomal recessive disorder, the chances for the child to develop cystinosis are very low if the father is unaffected. However, since he could be an asymptomatic carrier, pre-conceptional genetic counselling should be offered ,17. If tCTNS gene. The delay of a few weeks in the diagnosis of cystinosis is unlikely to influence long-term prognosis, since the diagnosis will still be made very early, and treatment can be started accordingly.In our expert opinion, we do not advise performing prenatal genetic testing to exclude cystinosis in an unborn child given the risk of miscarriage caused by the procedure. If early genetic testing is warranted, DNA can be extracted from cord blood or from the baby after birth to perform urgent analysis of the Primary hypogonadism in male cystinosis patients was first described in 1993 . In thosReduced kidney function or immunosuppressive treatment for kidney transplantation did not seem to be the cause for these hormone disturbances ,19,20. IIn the early cohort described by Chik et al., who were likely to be treated with cysteamine less vigorously compared to current practice, testicular volume was reduced in all 10 studied men . In moreThere are sporadic reports of testicular biopsy samples, the first being a postmortem investigation showing fibrosis, germinal dysplasia and Leydig cell hyperplasia with numerous cystine crystals, but seminiferous tubules were still visible . Later, In addition, the infiltration of activated macrophages was found in the interstitial testicular tissues, as well as the presence of perturbed blood–testis barrier in infantile cystinosis patients using histological analysis and Zonula occludens-1 as a marker for the quality of the blood–testis barrier. This could indicate that inflammation might be a common cause for both the primary hypogonadism and for epididymal dysfunction, ultimately causing obstruction .In the first paper that mentions semen analysis in cystinosis patients, azoospermia was found in all three investigated men, including one patient with normal sex hormone levels .Years later, semen analysis in five male cystinosis patients confirmed azoospermia in all subjects, even in those with normal sex hormone levels, and normal ejaculate volumes and pH in four of them. In this report, for the first time, concerns were raised regarding male fertility in cystinosis patients treated with cysteamine .CTNS mutation did not seem to play a role, since all four men harbored severe mutations in CTNS [Later studies confirmed azoospermia in most men suffering from infantile cystinosis; however, oligozoospermia was found in 1 out of 10 male infantile cystinosis patients in a subsequent investigation  and anot in CTNS ,21. Even in CTNS .Since male cystinosis patients with a normal Johnson score in their testicular biopsies were often reported to also suffer from primary hypogonadism, this primary hypogonadism could not have been the cause of the observed azoospermia. On the other hand, no sperm could be retrieved by percutaneous epididymal sperm aspiration (PESA) on several occasions in a man with a normal Johnson score, which could indicate a nonobstructive due to his azoospermia. Since testicular ultrasound also showed no signs of obstruction in two additional men, a nonobstructive cause of the azoospermia was initially suspected . HoweverSince the finding that there seemed to be no signs of obstruction on testicular ultrasound in two men with cystinosis , it has Several markers in seminal plasma have been studied in order to confirm an obstructive due to the azoospermia observed in infantile cystinosis. One study aimed to perform semen analysis in 15 male infantile cystinosis patients, although semen volume was too small (<0.3 mL) in two of them for seminal markers to be determined. In the remaining patients, normal levels of neutral α-glucosidase (NAG), secreted mainly by epididymis, were found in all except one patient, and the authors concluded that the epididymal secretory capacity and flux of secretions was not affected. They found reduced levels of both fructose , and zinc in 33% of the studied patients. The authors hypothesized that reduced fructose levels could be a sign of obstruction at the level of the excretory ducts of the vesicular glands and that reduced zinc levels could represent an obstruction at the level of the prostate. These abnormalities, however, did not explain all cases of azoospermia or oligozoospermia, since the remaining 67% of patients had normal levels of these seminal plasma markers .In another study, the seminal plasma levels of the epididymal secreted markers Extracellular Matrix protein-1 (ECM-1), which is highly expressed in the epididymis, and NAG, which is a specific and established marker for epididymal secretion used in the previous study as a seminal marker , were anCtns−/− knockout mouse model [Ctns gene with an IRES-βgalneo cassette [Ctns−/− knockout mice showed normal fertility compared to their wild-type litter mates, represented by a similar litter size, testicular morphology, and semen analysis. This meant that the knockout model, albeit showing a renal but mild phenotype, cannot be used to investigate male fertility in cystinosis in absence of the reproductive phenotype [Ctns−/− knockout mice had a perturbed blood–testis barrier [−/−Ctns knockout mice, as well as the reproductive phenotype, is less pronounced compared to humans.Unequivocally, animal studies can be of great importance to understand mechanisms and pathophysiology of diseases. In this context, male fertility in cystinosis was studied using a se model  that wascassette . The reshenotype . It was henotype ,35,36. L barrier . A possiCtns−/− zebrafish were fertile. However, female Ctns−/− zebrafish showed reduced egg production and percentage of fertilized eggs compared to wild-type zebrafish [In an attempt to find a more suitable animal model for cystinosis, zebrafish were proposed as an alternative ,38. In aCTNS gene was downregulated by siRNA silencing [In addition to these studies on men and mice, a human epididymal epithelial cell line  was studilencing . A transilencing . Since tilencing .Cysteamine is a cystine-depleting agent and the only available lifesaving treatment for nephropathic cystinosis. In early studies, it was proven to be a protective agent for spermatogonial stem cells (SSCs) in rats, following X-ray irradiation ,41. In aIn contrast, there are no clear studies on the effects of cysteamine on sperm quality in men. It had been hypothesized that cysteamine could, in theory, cause increased ghrelin levels, which in turn could have a negative effect on Leydig and Sertoli cell function . HoweverCtns−/− knockout mice that were fed with food mixed with cysteamine at a high dose of 500 mg/kg/day for 6 months [Recently, the effect of cysteamine in the previously described cystinosis mouse model was studied, using wild-type C57BL/6 mice and 6 months . FollowiIn accordance, in a previous study investigating the effect of oral cysteamine on female reproduction in wild-type rats, the authors showed that cysteamine had no adverse effects on conception and early embryonic development . Hence, Even though female infantile cystinosis patients seem to have normal fertility, there is room for future research to focus on neonatal outcomes in these children. To our knowledge, there are no reports of intrauterine growth retardation caused by placental dysfunction in cystinotic women, although in theory placental cystine accumulation could adversely influence its function. However, this has never been studied in a systematic manner. Additionally, it would be interesting to study whether or not compliance to cysteamine treatment prior to pregnancy improves pregnancy outcomes.Given the obstructive origin of azoospermia, preserving gonadal function in male cystinosis patients, if possible, is of the utmost importance to safeguard the possibility to father their own offspring via assisted reproductive technology. Therefore, future research should be aimed at further elucidating the exact pathogenesis of hypogonadism, which is now presumed to be related to inflammation and fibrosis caused by cystine deposition. More specifically, studying Leydig and Sertoli cell function in cystinosis and how to preserve it, could be of interest. In addition, it remains unclear why regular cysteamine dosages have subtherapeutic effects in testicular tissue and future studies could focus on different treatment strategies that have more of an effect on testicular cystine accumulation. In this regard, a cystinosis animal model more resembling human disease and in vitro models are indispensable.Additionally, it would be interesting to investigate the inflammatory aspect of cystinosis and its role in the reproductive phenotype, since this could open the door to study the use of anti-inflammatory agents to alleviate the reproductive symptoms. In addition, the use of medications that can lower the seminal plasma viscosity has not been studied thus far.Furthermore, semen analysis and sperm cryopreservation should be offered to all post-pubertal cystinosis males and hence be included as a recommendation in the standard clinical management guidelines for cystinosis. Once implemented, a large-scale registry-based observational study could provide the most reliable data on the proportion of infantile male cystinosis patients that show oligo- instead of azoospermia and give a better insight into their clinical determinants.CTNS mutation.In conclusion, female cystinosis patients seem to have a normal fertility, although complications during pregnancy that are secondary to the disease and kidney transplantation frequently occur. In contrast, most male infantile cystinosis patients suffer from azoospermia due to obstructive causes, with intact spermatogenesis in early adulthood. This renders them able to father their own biological children using epididymal or testicular sperm, followed by ICSI. Given the fact that cystinosis is an autosomal recessive disorder, the chances for the child to develop cystinosis are extremely small when the partner is confirmed not to be carrier of a Surprisingly, oligozoospermia or even normozoospermia can be found sporadically in male infantile cystinosis patients (mainly in those <30 years of age), which may be due to the presence of another yet unidentified factor that mitigates the effect of cystinosis on fertility. Moreover, oligozoospermia and normozoospermia have been reported in juvenile and ocular cystinosis, respectively. Based on this specific finding, we recommend performing a semen analysis at the earliest age possible for  male cystinosis patients to confirm the presence or absence of azoospermia, since some patients could still have a few sperm cells in their semen which could be cryopreserved. This will increase their opportunities to father their own biological children. Importantly, compliance to cysteamine treatment does not negatively affect the clinical fertility phenotype in male cystinosis patients, albeit it is not able to restore fertility.


International Journal of Neuropsychopharmacology.Misiak, B., Samochowiec, J., Konopka, A., Gawrońska-Szklarz, B., Aleksander Beszłej, J., Szmida, E., and Karpiński, P. (2021) Clinical Correlates of the NR3C1 Gene Methylation at Various Stages of Psychosis. In the originally published version of this manuscript, Dr Paweł Karpiński’s affiliation was incomplete. The correct affiliation is as follows: Department of Genetics, Wroclaw Medical University, Wroclaw, Poland and Laboratory of Genomics & Bioinformatics, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland. This has now been corrected online.


Bemisia tabaci affects plant performance by feeding directly from its energy sources, making it difficult to quantify crop damage, which is indirectly assessed through losses in productivity. The goal of this study was to characterize the influence of B. tabaci feeding on soybean, an economically valuable crop, and attempt to identify the optimal parameter for direct quantification of crop damage. A correlation network was created to extract biological interactions between the plant and B. tabaci nymphs at different densities and plant stages. Nymphs were more abundant during the vegetative stage, and a strong correlation between the density of nymphs and starch and fructose content was observed. The photosynthetic parameter turn-over number N was positively correlated with nymph density at a low-infestation level and negatively correlated with nymphs when they occur at a high infestation level. This association between nymph density and N may allow for development of a ranking scale to predict pest density, representing a useful tool for evaluating the potential impact of B. tabaci on soybean, especially in large areas, where nymph monitoring can be time-consuming.Bemisia tabaci (MEAM1) represents a species of economic importance in soybean. One of the obstacles to the management of B. tabaci is the quantification of damage by the pest because damage is indirectly inferred through losses in productivity. The objective of this study was to characterize the influence of B. tabaci feeding on soybean by assessing effects on photosynthetic parameters and the sugar and starch content of soybean leaves. The goal was to identify the optimal parameter to directly quantify pest damage on crop yield. Correlation networks were created among data on sugar content , starch and photosynthetic parameters , and the number of nymphs at each of three infestations level  during both the vegetative and reproductive stage of the crop. In general, nymphs were more abundant during the vegetative stage. Starch content was strongly correlated with nymph density. A strong positive correlation was observed between fructose and nymph density during the vegetative stage. Among the photosynthetic parameters, the turn-over number N was positively correlated with nymph density at a low-infestation level and negatively correlated with nymphs when they occurred at a high-infestation level. B. tabaci feeding affected the plant’s physiology and its interaction is reflected in part by the relationships among photosynthetic parameters as well as the levels of sugars and starch. This understanding might be useful in developing better monitoring tools for pest management. Bemisia tabaci MEAM1 (Gennadius 1889) (Hemiptera: Aleyrodidae), is considered one of the most destructive invasive pests of many crops, including soybean (Glycine max (L.) Merril) [Capnodium sp.) that causes sooty mold due to the excretion of honeydew.The whitefly,  Merril) , responsB. tabaci is the ability to quantify the damage caused by the pest, as these are indirectly inferred through losses in productivity and are invariably confounded by the impacts of other pests during the soybean production cycle. An alternative to directly assessing the damage caused by B. tabaci is to estimate the effects of its feeding on photosynthesis and the sugar and starch composition of the leaves.One of the obstacles to the management of B. tabaci [B. tabaci development [B. tabaci and might be useful for understanding the influence of B. tabaci on soybean at different phenological stages under different pest population densities.The interaction between the chemical composition of leaves and the performance of whiteflies has been investigated in eggplant, and results show that varieties with higher glucose and sucrose concentrations provide better conditions for the development of . tabaci . A posit. tabaci , and hig. tabaci . In soybelopment ,7. TheseB. tabaci in soybeans is by assessing how infestations interfere with photosynthesis. Reductions in photosynthetic rate caused by B. tabaci infestations has been reported for many crops, including tobacco, eggplant, zucchini, and cotton [B. tabaci feeding has been associated with a reduction in the net photosynthetic rate in zucchini [B. tabaci causes damage to soybean [B. tabaci feeding on soybean by assessing effects on photosynthetic parameters, chlorophyll, and the sugar and starch content of soybean leaves, with a goal of establishing better parameters to quantify crop damage by this pest.Another way to investigate the injury caused by d cotton ,9,10,11.zucchini . It is k soybean , resultiGlycine max (L.) Merril) as the host. Plants were grown in 2-L pots in a greenhouse and replaced every 20 days. The initial whitefly population was collected from a cotton field at the United States Department of Agriculture—Arid Land Agricultural Research Center (USDA-ALARC) in Maricopa, Arizona, USA, and was previously identified as B. tabaci biotype B (MEAM1). Molecular characterization of the insects was made periodically during the study for confirmation of the insect biotype [Whiteflies were reared in cages in a greenhouse using soybean ( biotype .var. 01057408) was sown in seed trays containing a general-purpose growing medium. When the seedlings reached the V1 phenological stage [B. tabaci. Each plant was covered with an anti-aphid screen attached to the bottom of the pot that served as a cage for eventual whitefly infestation. The experiment was carried out under greenhouse conditions.Soybean . Then, the leaf area of the sample was measured with a LI-3100C .For the Chlorophyll Fluoresce Induction Kinetics (OJIP) protocol, a leaf on the plant was dark-adapted for at least 20 min with detachable leaf clips. Measurements were then performed using the FluorPen FP100 .0), performance index on absorption basis (Pi_Abs), and turn-over number (N). These assays were performed in situ in the greenhouse weekly.A clip was attached to one of the leaflets in the upper middle third of the plant before the measurements. The device emits a saturation pulse through a beam of light, which when reflected, is read by the device. The OJIP protocol is a dark-adapted chlorophyll fluorescence technique used to measure plant stress. The parameters measured include initial fluorescence  and chlorophyll .The leaflets were individually homogenized in liquid nitrogen using a pestle and a mortar, weighted in two aliquots of 20 mg each sample, placed in 2-mL EppendorfAn assessment of the quantity and quality of sugars and starch present in the infested soybean leaves was carried out for all the leaf samples previously prepared. The evaluation of the sugar content was carried out by first preparing the samples for extraction of non-structural carbohydrates and then extracting the starch for subsequent chromatographic analysis.A total of 25 µL of maltose (20 mg/mL of 80% ethanol) and 1 mL of 80% ethanol were added to the tubes containing the samples; the tubes were placed in a water bath at 85 °C for 10 min and then centrifuged for 5 min at 10,000 rpm. The supernatant was transferred to a new 2-mL tube. Then, 0.5 mL of 80% ethanol was added to the tubes containing the precipitate, submitting them again to the water bath and centrifugation as described above two more times, adding the supernatants each time. The tubes containing the total supernatant were adjusted to 2 mL with 80% ethanol and centrifuged once more for 5 min at 10,000 rpm. The samples were then transferred to HPLC vials using a syringe and a 0.45-µm filter. The sugars assessed were fructose, glucose, and sucrose. Maltose could not be analyzed because the test protocols for other sugars required the addition of maltose, which was used as a surrogate.Then, 25 µL of maltose (20 mg/mL in 80% ethanol) and 500 µL of 2M potassium hydroxide were added to the tubes containing the centrifuged precipitate resulting from the extraction of non-structural carbohydrates, placed in a water bath at 100 °C for 10 min, and then allowing to cool to room temperature. Then, 100 µL of 1M acetic acid (PH = 4.5), 50 µL of Tris 1M buffer solution (PH = 7.2), and 100 µL of alpha-amylase were added. The tubes were placed in a water bath at 85 °C for 30 min and allowed to cool to room temperature. A total of 100 µL of 1M acetic acid (PH = 4.5) and 500 µL of amyl glucosidase were added, and again, tubes were placed in a water bath at 100 °C for 4 min. Finally, 75 µL of 2M sodium hydroxide was added and the volume of the tubes adjusted to 2 mL with 100% ethanol for centrifugation at 6000 rpm for 5 min. The sample solution was then transferred to HPLC vials using a syringe and a 0.45-µm filter.The analyses of content were determined in 2 mL of each sample following methods proposed for measurements of nonstructural carbohydrates in plant tissues  using a ® tubes containing the samples previously weighted. The tubes were completely covered with aluminum foil to avoid exposure to light and placed on a shaking plate in a refrigerator at 4 °C for 48 h, inverting the tubes twice a day to keep the samples homogeneous. After 48 h, the samples were centrifuged at 10,000 rpm for 10 min, then 200 µL of the supernatants were transferred to propylene microplates to perform the readings.For the extraction of chlorophyll, 1 mL of 100% methanol at 4 °C was added to the 2-mL Eppendorf® HT microplate reader . The absorbances were then used in the calculations of chlorophyll a, b, and a + b by multiplying the absorbance values by the respective correction factor (A665.2 = 0.478) or (A652 = 0.466) divided by the sample weight (mg) to obtain the value in µg/mg using the following equations :The chlorophyll contents were obtained by spectrophotometry at absorbances 652 and 665, performed with the SynergyStatistical analyses were performed using the R Core Team  program.p > 0.05, In the univariate analyses, we observed no significant interactions between infestation treatment and plant growth stage for any of the variables. We also found no differences between treatments or stages  in two phenological stages, vegetative and reproductive. The more solid the color of the line, the stronger the correlation between the factors. Correlation coefficients and significance levels are provided in The number of nymphs was significantly different between the treatments both in the vegetative and reproductive stages, and we observed higher numbers as infestation levels increased. Differences in leaf area were observed only during the vegetative stage .B. tabaci, they are not included in this correlation network analysis.The objective of the correlation network analysis was to observe how the interactions between the factors evaluated were altered by different densities of nymphs. Because the control treatments were not infested with ρ = 0.99; p < 0.01)   . This co < 0.05) . The sam < 0.05) .ρ = −0.92; p < 0.01) was negative; however, in the vegetative stage, these correlations were positive . This same pattern was observed between fructose and nymphs. The networks also show that Pi_Abs and fructose are positively correlated in all the treatments.In the reproductive stage, the correlation of nymphs and Pi_Abs   (We observed different patterns of correlation between the reproductive and vegetative stages . In the  < 0.05) . In the  > 0.05) .ρ = −0.16; p > 0.05) and a positive correlation between nymphs and starch  .While phenological stages altered the correlation network, the different infestation treatments also affected the relationships between the parameters in the correlation network. For medium infestation in the reproductive stage, we observed a weak negative and not significant correlation between nymphs and fructose ( < 0.05)  and betwρ = 0.75; p < 0.05) and starch  were positive  and also a strong positive correlation between nymphs and starch  , but again no significant correlation was observed with Pi_Abs  , which i < 0.05) . For phoh Pi_Abs .ρ = 0.93; p < 0.01) and sucrose  and moderate and non-significant for starch . No significant correlation was observed for glucose   and F0 negatively correlated , while N was again negative , following the same pattern described for the low-infestation treatment.In the high-infestation vegetative stage treatment, the correlations of nymphs with sugars were positive for fructose ( > 0.05) . For phoa content between plants in the vegetative and reproductive stages. A higher concentration in the vegetative stage was observed for all levels of infestation although there was no difference between treatments in the same plant phenological stage  likely affected leaf growth, as was reflected in reduced leaf area  from a photosynthetic organ like a leaf should be lower compared with treatments that subject the plant to some type of stress [B. tabaci induced stress that was measurable through changes in chlorophyll fluorescence. While there was a positive correlation between nymphal density and F0 at both low- and high-infestation rates, there was a negative correlation for medium infestations. The same pattern of correlations was observed for starch. This is consistent with a reduction in chlorophyll b content in the medium infestation during the reproductive stage. Previous studies have shown that increases in chlorophyll b content results in increased amount of starch content, both indicators of growth vigor in plants [0, which is one most representative chlorophyll fluorescence parameters and widely used in studies to measure stress on plants [Ideally, in a healthy plant, the initial fluorescence ).For the absorption flow performance index (Pi_Abs), the expected response is the inverse of that expected for Fhe plant . TreatmeA) was reduced to QA- in the interval from time zero (t0) to reach maximum fluorescence (Fm) after receiving electrons from water using light energy. This direct electron transfer is essential in the process of converting light energy into chemical energy [A) such that the plant would be absorbing more light but not converting this light energy to photoassimilates. Previous study has shown that increases in F0, which is inverse to Pi_Abs, represents more QA- formation [0 and nymphs. Higher levels of chlorophyll (a + b) were observed in the vegetative treatments, and during the reproductive treatments, the medium infestation showed the lowest concentration. We also observed the inversion in the correlations between nymphs and starch and between nymphs and F0 in the medium-infestation-level treatments. In other words, besides a higher light absorption, the plant also depends on good photosynthetic performance to provide resources such as starch for plant health, making it a more suitable host to B. tabaci.The N, or turn-over number, represents how many times the primary quinone acceptor (Ql energy ,27. An iB. tabaci infestation and photosystem activity is expected. Here, that situation resulted in poorer plant development, as indicated by smaller leaves in soybean under high insect infestation.Whiteflies are phloem-feeders, and phloem is responsible for transporting photoassimilates, with generally high concentrations of sugars . The priB. tabaci development. Glucose is second only to maltose as a product of starch breakdown and a source of carbon exported from chloroplasts during its degradation [The vegetative stage appears to provide better conditions for the insect’s development, as evidenced by the faster growth of nymphs during this stage. Differing from that observed for the other sugars, the correlations between glucose with nymphs were non-significant, indicating that during the vegetative stage other components, such as fructose and starch, may provide better conditions for radation .Even though glucose also is a product of starch breakdown, it did not present negative correlation with starch. First, it could be that maltose provided an energetic advantage over the export of glucose, representing two-thirds of the carbon export, while glucose represents the rest . SecondlB. tabaci nymphs occurred. A similar pattern was observed for sucrose. Varieties with higher content of fructose and sucrose in their leaves were reported to promote higher densities and better conditions for the development of B. tabaci in eggplants and cotton, respectively [B. tabaci needs during the reproductive stage.One of the main components of honeydew excreted by whiteflies is fructose ,32, whicectively . On the ectively . During ectively ,7. This B. tabaci in the crop, especially in large areas, where nymph monitoring can be time consuming.Among the photosynthetic parameters, the turn-over number N was positively correlated with nymphs at lower densities of nymphs and negatively correlated with nymphs at higher densities. These correlations also were affected by the phenological stage, whereby correlations were negative during the vegetative and positive during the reproductive stage. Nevertheless, this pattern is modified somewhat by the density of infestation. During the reproductive stage, the correlation is strongest at the lowest density of infestation and declines in strength with increasing density. During the vegetative stage, the negative correlation between nymphs and N is weakest at the lowest infestation and grows in strength as density increases at the medium infestation. Although the maximum correlation was not observed at the high infestation compared with the medium infestation, a strong negative correlation was still observed. This may have been influenced by the significant drop in chlorophyll content observed at the high infestation affecting N. The assessment of the correlation between nymphs and N may allow for development of a ranking scale of N values to predict density of nymphs and might represent a useful tool for evaluating the impact of B. tabaci feeding affects the plant’s physiology reflected in part by the relationships among photosynthetic parameters as well as the levels of sugars and starch. From our results, the greatest impacts of insect infestation occur during the vegetative phenological stage, when plants are more suitable for B. tabaci development, thus leading to plant stress that will likely have consequences on productivity.


The dimensional error of forming the architecture was 1% as a coefficient of variation. Bone marrow-derived osteoblasts were cultured on a polished surface and on meso- and nano-scale hybrid textured surfaces with different spike heights. The osteoblastic differentiation was significantly promoted on the hybrid-textured surfaces compared with the polished surface, and among them the hybrid-textured surface with 40 μm-high spikes showed unparalleled performance. In vivo bone-implant integration also peaked when the hybrid-textured surface had 40 μm-high spikes. The relationships between the spike height and measures of osteoblast differentiation and the strength of bone and implant integration were non-linear. The controllable creation of meso- and nano-scale hybrid biomimetic surfaces established in this study may provide a novel technological platform and design strategy for future development of biomaterial surfaces to improve bone integration and regeneration.Biomimetic design provides novel opportunities for enhancing and functionalizing biomaterials. Here we created a zirconia surface with cactus-inspired meso-scale spikes and bone-inspired nano-scale trabecular architecture and examined its biological activity in bone generation and integration. Crisscrossing laser etching successfully engraved 60 μm wide, cactus-inspired spikes on yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) with 200–300 nm trabecular bone-inspired interwoven structures on the entire surface. The height of the spikes was varied from 20 to 80 μm for optimization. Average roughness (Sa) increased from 0.10 μm (polished smooth surface) to 18.14 μm (80 μm-high spikes), while the surface area increased by up to 4.43 times. The measured dimensions of the spikes almost perfectly correlated with their estimated dimensions (R Bio-inspired or biomimetic design of biomaterials presents new possibilities for developing implantable devices with enhanced biocompatibility and novel functions ,5,6,7,8.A recent laser technology advancement made a breakthrough in simultaneous creation of meso- and nano-scale topography on zirconia . ZirconiWe attempted to create zirconia surfaces with cactus-like meso-scale spikes and nano-scale architecture using crisscrossing solid-state laser etching, as illustrated in We continued analyzing the meso-/nano-scale hybrid textured zirconia. Three-dimensional images captured 3-D projection of the meso-scale spikes with heights incrementally increased from 20 to 80 μm A. Zoomed2 = 0.9975; Cross-sectional profiling along the line connecting spike peaks confirmed smooth, oscillating curves of spikes and valleys A. The wiIn a quest to optimize a zirconia surface for the promotion of bone–implant integration, we cultured osteoblasts on meso-/nano-scale hybrid textured zirconia with various spike heights as well as on a polished-smooth zirconia surface. The number of osteoblasts attached to the zirconia surfaces was evaluated during the initial stage of culture on day 2. All hybrid-textured zirconia surfaces showed a significantly lower number of attached cells than on the polished surface , and amoWe then examined the ability of the zirconia samples to support cell proliferation by measuring the density of cells on day 5 of culture . The celWe next examined osteoblastic differentiation by measuring ALP activity, matrix mineralization ability, and the expression levels of osteoblastic differentiation marker genes. Unlike the results in the cell attachment and cell density assays, the ALP activity measured on day 10 was significantly higher for all meso-/nano-scale hybrid textured zirconia than on the polished surface  and cylindrical form , prepared from yttria-stabilized tetragonal zirconia polycrystal (Y-TZP), were polished and assigned as the “polished-smooth surface” group. To create dual-scale textured surfaces, polished zirconia samples were treated with solid-state laser etching. The laser etching was established to carve grooves with hemispherical bottom surfaces [−8 M dexamethasone, and antibiotic–antimycotic solution containing 10,000 units/mL penicillin G sodium, 10,000 mg/mL streptomycin sulfate, and 25 mg/mL amphotericin B. Cells were incubated in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. At 80% confluency, the cells were detached using 0.25% trypsin–1 mM EDTA-4Na and seeded onto zirconia disks placed in 12-well culture dishes at a density of 3 × 104 cells/cm2. The culture medium was renewed every three days.Bone marrow-derived osteoblasts were isolated from the femurs of 8-week-old male Sprague–Dawley rats and placed into alpha-modified Eagle’s medium supplemented with 15% fetal bovine serum, 50 μg/mL ascorbic acid, 10 mM Na-ß-glycerophosphate, 10The number of osteoblasts attached to the zirconia surfaces during the initial stage of culture was evaluated using a tetrazolium salt (WST-1)-based colorimetric assay  on day 2 of culture as described elsewhere ,34,45,602O and 250 μL of p-nitrophenyl phosphate was added, followed by incubation at 37 °C for 15 min. ALP activity was evaluated by measuring the released nitrophenol in the enzymatic reaction and determined at 405 nm using a plate reader (Biotek). For staining, cultured osteoblasts were washed twice with Hanks’ solution and then incubated with 120 mM Tris buffer (pH 8.4) containing 0.9 mM naphthol AS-MX phosphate and 1.8 mM fast red TR for 30 min at 37 °C.Osteoblast ALP activity was examined on day 10 using two methods of chemical detection: colorimetry and a staining-based assay. As previously described ,27,61, tThe mineralization capability of cultured osteoblasts was examined by colorimetry-based quantification of calcium deposition on day 15. The cultures were washed with PBS and incubated overnight in 1 mL of 0.5 mM HCl solution with gentle shaking. The solution was mixed with o-cresolphthalein complexone in an alkaline medium  to produce a red calcium cresolphthalein complexone complex. Color intensity was measured by an ELISA plate reader at 575 nm absorbance.Gapdh expression was used as the endogenous control.Gene expression was analyzed using qPCR on days 5, 10, and 15. Total RNA was extracted from cells using TRIzol  and a Direct-zol RNA MiniPrep kit . Extracted RNA was reverse transcribed into first-strand cDNA using SuperScript III Reverse Transcriptase (Invitrogen). Quantitative PCR was performed in a 20 μL volume containing 90 ng cDNA, 10 μL TaqMan Universal Master Mix II, and 1 μL TaqMan Gene Expression Assay using a QuantStudio 3 Real-Time PCR System , to quantify the expression of type I collagen and osteocalcin mRNA. Eight-week-old male Sprague-Dawley rats were anesthetized with 1–2% isoflurane inhalation. After their legs were shaved and scrubbed with 10% povidone-iodine solution, the distal aspects of the femurs were carefully exposed via skin incision and muscle dissection. The flat surfaces of the distal femurs were selected for implant placement. The implant site was prepared 9 mm from the distal edge of the femur by drilling with a 0.8 mm round burr and enlarged using reamers (#ISO 090 and 100). Profuse irrigation with sterile isotonic saline solution was used for cooling and cleaning. One of the variously textured zirconia implants (1 mm in diameter and 2 mm in length) was placed into a prepared hole. Surgical sites were then closed in layers. Muscle and skin were sutured separately with resorbable suture thread. The University of California at Los Angeles (UCLA) Chancellor’s Animal Research Committee approved this protocol, and all experimentation was performed in accordance with the United States Department of Agriculture (USDA) guidelines on animal research .The established implant biomechanical push-in test was used to assess the biomechanical strength of bone–implant integration ,24,25. AAfter the push-in test, implants were carefully exposed and soaked in agitated water for one hour and dried under heat and vacuum. After being carbon sputter-coated, the specimens were examined by scanning electron microscopy (SEM). The elemental composition of the tissue remnants and the implant interface were analyzed by energy dispersive X-ray spectroscopy (EDX) .n = 6). Three disks were used for all cell culture studies (n = 3). Six implants were used for the biomechanical push-in test (n = 6) for each of the implant surfaces at each healing time point. One-way ANOVA was performed to examine the differences among differently textured surfaces. When appropriate, Bonferroni’s test was used as a post-hoc test. p-values less than 0.05 were considered statistically significant.Data on surface roughness parameters were collected from six sites on three different disks (We have created a meso-/nano- dual-scale bio-inspired zirconia surface by laser etching. The surface carries cactus-inspired meso-scale spikes and bone-inspired nano-trabeculae. The height of the meso-spikes is controllable and in this study was varied incrementally from 20 to 80 μm. The function of osteoblasts and the bone-implant integration responded in a non-linear fashion to the spike height and peaked or plateaued when the spikes were 40 μm high. These 40 μm-high meso-spikes increased Sa to 7.72 μm from the baseline level of 0.10 μm for a polished-smooth zirconia surface, and likewise increased the surface area by 2.64 times. The dual-scale bio-inspired texturing was proven feasible on the complex surface of a prototype dental implant, providing a novel, biomimetic platform technology and design strategy for developing dental and orthopedic implants and other biomaterials.


Philip Alexander, MD, is a native Texan, retired physician, and accomplished musician and artist. After 41 years as an internal medicine physician, Dr. Phil retired from his practice in College Station in 2016. A lifelong musician and former music professor, he often performs as an oboe soloist for the Brazos Valley Symphony Orchestra. He began exploring visual art in 1980, evolving from pencil sketches—including an official White House portrait of President Ronald Reagan—to the computer-generated drawings featured in this journal. His images, which first appeared in this journal in the spring of 2012, are his own original creations. To learn more about Dr. Phil and his art, read “Art and Medicine with Dr. Phil” in issue 13.2.Methodist DeBakey Cardiovascular Journal, submit your creation online at journal.houstonmethodist.org as a “Humanities” entry.If you would like to see your art published in the


Acinetobacter has been frequently detected in backwater areas of the Three Gorges Reservoir (TGR) region. We here employed Caenorhabditis elegans to perform biosafety assessment of Acinetobacter strains isolated from backwater area in the TGR region. Among 21 isolates and 5 reference strains of Acinetobacter, exposure to Acinetobacter strains of AC1, AC15, AC18, AC21, A. baumannii ATCC 19606T, A. junii NH88-14, and A. lwoffii DSM 2403T resulted in significant decrease in locomotion behavior and reduction in lifespan of Caenorhabditis elegans. In nematodes, exposure to Acinetobacter strains of AC1, AC15, AC18, AC21, A. baumannii, A. junii and A. lwoffii also resulted in significant reactive oxygen species (ROS) production. Moreover, exposure to Acinetobacter isolates of AC1, AC15, AC18, and AC21 led to significant increase in expressions of both SOD-3::GFP and some antimicrobial genes  in nematodes. The Acinetobacter isolates of AC1, AC15, AC18, and AC21 had different morphological, biochemical, phylogenetical, and virulence gene properties. Our results suggested that exposure risk of some Acinetobacter strains isolated from the TGR region exists for environmental organisms and human health. In addition, C. elegans is useful to assess biosafety of Acinetobacter isolates from the environment. That is, a novel ecosystem is created by construction of the TGR Dam. Meanwhile, due to rapid development in industrialization and urbanization in the recent years, various organic and inorganic pollutants are potentially released into the environment in the TGR region through industrial or residential wastewater5. Mover, a large amount of bacterioplankton community, including waterborne pathogens, has been detected in the TGR region8.The Three Gorges Reservoir (TGR), whose distance is approximately 662.9 km, is a major water source in China. Water fluctuation in the TGR region produces a water-level fluctuating zone (WLFZ) every yearCaenorhabditis elegans can be used to perform the toxicological study at the whole animal level12. C. elegans has become an ideal surrogate model to determine both pathogenesis and conserved mechanisms in host-microbe interactions of human pathogens14. More importantly, C. elegans is highly sensitive to various environmental exposures18. Considering the sensitivity to environmental exposure, it has been employed to perform biosafety evaluation of water samples in TGR region in both flood season and quiet season20. Based on our previous toxicity evaluation, only acute exposure to water sample in backwater area resulted in toxic effects on nematodes, such as decrease in locomotion behavior and activation of oxidative stress20. Moreover, both liquid phase and solid phase contributed to toxicity induction of water sample in backwater area20. In the liquid phase, the potential toxicants were suggested to be the organic pollutants20. Nevertheless, the toxicity contributors in the solid phase of water sample in backwater area in the TGR region are still largely unclear.8. Environmental Acinetobacter was realized as an important nosocomial pathogen during the late 1970s, and has received an increasing attention because of its potential to cause severe nosocomial infections and formation of multiple-drug and pan-drug resistance22. Acinetobacter occupies a considerable position in nature because it prevails in natural environments, such as soil, oceans, fresh water, and sediments24. Nevertheless, the biosafety properties of Acinetobacter in the TGR region remain largely unclear. We here aimed at performing biosafety assessment of Acinetobacter strains isolated from backwater areas in the TGR region in nematodes. Among the isolated 21 Acinetobacter strains, exposure to four isolates  resulted in toxic effects in nematodes. Our data implied the possible exposure risk of some Acinetobacter strains in the TGR region for environmental organisms and human health.Environmental pathogens are an important component in the solid phase of surface water samples. Existence of high prevalence pathogens has been found in backwater areas of the TGR regionAcinetobacter strains  , A. junii NH88-14 (FJ447529), A. baumannii ATCC19606T (HE651907), A. lwoffii DSM2403T (X81665) and A. haemolyticus TTH04-1 (KF704077) . Among the named species, A. baumannii is the most studied species associated with clinical infections followed by the non-A. baumannii species A. haemolyticus, A. junii, A. johnsonii, and A. lwofii21.In the TGR region, we isolated 21 s) Table . Based onk Table . In phylAcinetobacter strains for 24-h. Totally 21 Acinetobacter strains isolated from the TGR region and 5 reference strains of Acinetobacter species were used for the lifespan analysis. Based on the comparison of lifespan curves, exposure to Acinetobacter strains of AC2, AC3, AC4, AC5, AC6, AC7, AC8, AC9, AC10, AC11, AC12, AC13, AC14, AC16, AC17, AC19, AC20, A. johnsonii H10, and A. haemolyticus TTH0-4 could not alter lifespan curve  different from that in control nematodes  and these 5 reference speices are important clinical microorganisms38, and A. baumannii ATCC 19606T is a model strain of pathogenic bacteria causing nosocomial infection39, followed by the non-A. baumannii species A. haemolyticus, A. junii, A. johnsonii, and A. lwofii21.The high prevalence pathogens exist in the backwater area of the TGR region20. In the liquid phase, the potential toxicants were suggested as the organic pollutants20. In this study, using lifespan as the toxicity assessment endpoint, we found that four  of the isolated and examined Acinetobacter strains and tree reference strains of A. baumannii, A. junii, and A. lwoffii significantly reduced lifespan  with oxidative stress activation. Nevertheless, we did not detect the decrease in SOD-3::GFP expression after exposure to the above pathogenic Acinetobacter strains. This may be largely due to the short exposure duration (24-h) for these pathogenic Acinetobacter strains. Usually, long-term exposure to toxicants at high concentrations causes decrease in SOD-3::GFP expression9. Exposure to nanopolystyrene (1–10 μg/L) caused increase in SOD-3::GFP expression, whereas exposure to nanopolystyrene (1000 μg/L) resulted in decrease in SOD-3::GFP expression40.We further observed the significant ROS production in nematodes exposed to fii Fig. A, which fii Fig. B, which Acinetobacter strains of AC1, AC15, AC18, and AC21 induced increase in expressions of some antimicrobial genes   was observed in nematodes infected with AC1, AC15, AC18, and AC21 . The reason to select these intestinal antimicrobial genes is that the ROS production is mainly activated in the intestine25. Similarly, we also did not observe the suppression in expressions of these antimicrobial genes in nematodes exposed to Acinetobacter strains of AC1, AC15, AC18, or AC21, which is also largely due to the performed short exposure duration (24-h) in nematodes. Moreover, we found that exposure to Acinetobacter strains of AC1, AC15, AC18, and AC21 induced the different dysregulation of examined antimicrobial genes  observed in mammals. Therefore, the further biosafety assessment experiments in mammals for the identified four Acinetobacter strains are still needed.In this study, we provide some lines of evidence to show the important value of Acinetobacter strains of AC1, AC15, AC18, and AC21. However, we did not observe the obvious difference in morphological properties of cell shape, arrangement of cell, Gram staining, and colony morphology among the examined Acinetobacter strains of AC1, AC15, AC18, and AC21  were collected from the depths of 0.5, 5, 10 m in the backwater area site. Water samples were used for the isolation of Acinetobacter after mixing fully in the sterile bucket, and water samples were stored at 0 °C after collection.The water sample was collected in backwater area  in Wanzhou, Chongqing in the flood season−1, and 10−2), inoculated into LB medium, and incubated at 37 ± 0.5 °C for 24 h. Subculture and purification of bacterial colonies were carried out by the streak plate method. For the purified bacterial isolates, genomic DNA of different bacterial isolates was extracted using bacterial genomic DNA extraction kit  according to the manufacturer’s instructions. The complete bacterial 16S rRNA gene was amplified with the primer set 27 F and 1492R. PCR products were visualized using 1% agarose gels stained with ethidium bromide. Positive amplicons were quantified using a PicoGreen dsDNA Assay kit . Purified products were sequenced and analyzed by Magigen . Phylogenetic tree was constructed using the Mega 5.0 program using the neighbor-joining (N-J) method with a 1000-bootstrap.The mixed water sample was diluted serially , A. lwoffii (DSM 2403T), A. junii (NH88-14), A. haemolyticus (TTH0-4), and A. johnsonii (H10) from China General Microbiological Culture Collection Center (CGMCC) were used to expose the Caenorhabditis elegans in this study. The information of these reference strains is listed in the Table Acinetobacter strains inoculated on broth agar medium were incubated for 24 h at 37 °C45. Primary identification and characterization of different Acinetobacter strains were performed to determine cell shape, arrangement of cell, gram staining, and colony morphology using UVsolo 2 touch 46. After the growth at 37 °C for 24 h, the biochemical properties of different Acinetobacter strains were further determined using standard Enterobacteriaceae biochemical identification tube 47.Different muIs84[SOD-3:GFP] and wild-type N2 were used. Normal nematode growth media (NGM) plates were used to maintain nematodes48. To prepare synchronized L4-larvae, gravid worms were first treated with bleaching solution (0.45 M NaOH and 2% HOCl)49. The released eggs were let to further develop into the L4-larvae population.CF1553/Acinetobacter strains. Different Acinetobacter strains were seeded on modified NGM containing 0.35% peptone. Exposure to different Acinetobacter strains was started by transferring nematodes onto each assay plate by adding 60 animals to each assay NGM plate. Full-lawn assay plate was used for Acinetobacter pathogenesis assay as described50. That is, the surface of assay NGM plate was all seeded Acinetobacter strains. The aim of using full lawn assay was to exclude the possibility of effect from the avoidance behavior of nematodes to Acinetobacter strains.The L4-larvae population was exposed to different 51. After the infection, the treated nematodes were transferred to M9 solution containing 25 mM levamisole to paralyze the nematodes. Nematodes were first transferred to a NGM plate containing ampicillin (1 mg/mL) and gentamicin (1 mg/mL) for 15 min to eliminate Acinetobacter on the body surface. Nematodes were further transferred to a new NGM plate containing ampicillin (1 mg/mL) and gentamicin (1 mg/mL) for 30 min to remove any residual Acinetobacter. After that, the nematodes were lysed with a motorized pestle. The lysates were serially diluted in M9 solution and plated on Luria–Bertani plates. After overnight incubation at 37 °C, the colonies were counted to determine the CFU per nematode. The experiments are repeated for three times.The method was performed basically as describedAcinetobacter strains for 24-h, the survival of worms was counted every day at 20 °C52. If no response was observed after prodding using platinum wire, the worms were considered as dead. The animals were transferred daily during the first 7-day. For the lifespan assay, 60 animals were examined for each treatment. Three replicates were carried out. We used log-rank test to analyze the lifespan curve data. Survival curves were considered to have significant difference if P values were ≤ 0.01.After exposure of L4-larvae nematodes to different 53. Body bend and head thrash were selected as the endpoints54. After exposure, the worms were first washed with M9 buffer. After that, assuming that animals traveled along x axis, a body bend is defined as a change of bending direction at the mid-body. A head thrash is defined as a change of posterior bulb direction along y axis. For each treatment, 40 animals were analyzed.Locomotion behavior reflects the functional state of motor neurons55. The method was performed as described56. After the exposure to different Acinetobacter strains, the animals were labeled for 3 h using CM-H2DCFDA (1 µM). After that, the animals were observed at 488 nm (excitation wavelength)/510 nm (emission filter) under a laser scanning confocal microscope. Using Image J software, we semi-quantified intestinal fluorescence intensity in comparison to intestinal autofluorescence. For each treatment, 50 animals were examined.Production of reactive oxygen species (ROS) reflects the activation of oxidative stresssod-3 encodes mitochondrial Mn-SOD9. Using Image J software, fluorescence intensity of SOD-3::GFP signals in the intestine was semi-quantified. For each treatment, 50 animals were examined.In nematodes, spp-1, lys-8, lys-7, lys-1, spp-12, dod-6, dod-22, and F55G11.4 were determined in real-time PCR system. The reference gene was tba-1. Three biological replicates were carried out. Primer information is provided in Table The total RNAs of control and exposed nematodes were extracted using Trizol. Using a spectrophotometer, concentration and purity of the obtained RNAs were determined. We performed the reverse transcriptase reaction with Mastercycler gradient PCR system for cDNA synthesis. With the aid of SYBR Green qRT-PCR master mix, transcriptional expression of Acinetobacter on GenBank. Primer information was provided in Table Primers were designed according to the published sequences of virulence genes of Statistical analysis was carried out using SPSS Statistics 19.0 Software . Probability level of 0.01 was considered statistically significant. Using one-way analysis of variance (ANOVA), the differences between groups were tested.Supplementary Information.


Both postprandial lipemia (PPL) and disturbed blood flow (DBF) induce endothelial dysfunction. However, the interactive effect of these stimuli on endothelial function is currently unknown. In the present study, we tested whether PPL plus DBF causes a greater reduction in flow-mediated dilation (FMD) than PPL and if this response is associated with elevations in oxidative stress and endothelial microvesicles (EMVs).2) randomly underwent two experimental sessions: PPL and PPL plus DBF. FMD and venous blood samples were obtained at baseline and 30, 70, and 110 min after stimulation. PPL was induced by fat overload via mozzarella pizza ingestion and DBF by forearm cuff inflation to 75 mm Hg per 30 min. Lipidic profile, oxidative stress  and EMVs were measured in blood samples.Eighteen individuals (aged 28 ± 1yrs, 3 females, and BMI 24.43 ± 0.8kg/m2O2 similarly increased in both sessions, but TBARS and FRAP did not change.Hypertriglyceridemia was observed in both sessions. Retrograde shear rate and oscillatory index responses were significantly higher in the PPL plus DBF compared with PPL. PPL plus DBF evoked a greater reduction in FMD than did PPL and EMVs, NADPH oxidase, and HThese data indicate that the association of PPL plus DBF additively impairs endothelium-dependent function in 110 min after stimulus in healthy individuals, despite a similar increase in oxidative stress and EMVs. Further studies are needed to understand the mechanisms associated with the induced-endothelial dysfunction by association of PPL and DBF. Coronary arterial disease is the main cause of death and reduction in quality of life worldwide, leading to enormous economic consequences . AtherosAn important risk factor for endothelial dysfunction is postprandial lipidemia (PPL) . The ingIt is well known that geometrically irregular arterial regions including bifurcations, branches, and curvatures, which are characterized by low time-average shear stress and high oscillatory blood flow profiles , are markedly susceptible to the development of atherosclerosis . These fEndothelial microvesicles (EMVs) are small vesicles released from the endothelium into the circulation in response to activation, injury, or apoptosis of endothelial cells . They haIn the present study, we hypothesized that short-term vascular exposure to PPL and DBF additively impairs endothelial function, as assessed by FMD, in young healthy subjects. We also attempted to determine whether this additive effect in the reduction of FMD induced by PPL and DBF is associated with increases in both circulating oxidative stress and EMVs.All experimental procedures and measurements conformed to the Declaration of Helsinki and were approved by the Research Committee of the Heart Institute (SDC 4549/17/049), the Human Subject Protection Committee at the Clinics Hospital of the University of São Paulo Medical School (CAAE 71981517.8.0000.0068). All participants provided written informed consent before enrollment.–1), aged 18–35 years, with no history of cardiovascular or metabolic diseases were included in this study. Initially, the participants were screened by telephone and reported to the laboratory following an overnight fast for a screening visit to verify eligibility. Exclusion criteria were as follows: systolic blood pressure ≥ 140 mm Hg, diastolic blood pressure ≥ 90 mm Hg, serum total cholesterol ≥ 200 mg dL–1; low-density lipoprotein-cholesterol ≥ 130 mg dL–1; high-density lipoprotein-cholesterol ≤ 35 mg dL–1; fasting glucose ≥ 100 mg dL–1. Screening for the study consisted of a complete medical history questionnaire, followed by a physical examination that included biochemical blood analysis, resting blood pressure (BP) measurement (sphygmomanometry), a resting electrocardiogram, and anthropometry .A total of 18 healthy subjects, non-smoking, recreational to highly active .The study protocol was performed randomly in two sessions, PPL and PPL plus DBF . All sesvia the pulsed-wave mode at linear frequencies of 13 and 6.0 MHz, respectively. The sample volume was located at the center of the brachial artery and then adjusted to cover vessel width. After this, baseline diameter and blood velocity waveforms were continuously recorded over 60 s. Reactive hyperemia was assessed immediately after relief of 5 min of total ischemia, which was induced by external compression of the arm using an inflatable cuff at 220 mm Hg. After this period, the cuff was rapidly deflated and the changes in artery diameters from baseline were expressed as percentages. The probe location was marked on the skin to guarantee that the FMD was repeated in the same location during the study.Brachial artery FMD was measured in the left arm, with the shoulder abducted at approximately 80° and the forearm supinated. In accordance with the most recent FMD guidelines , 2019, aPostprandial lipidemia (PPL) was induced through the ingestion of a mozzarella pizza (Sadia), with a total of 460 g and the following nutritional information: 59.4 g of fat, 102 g of carbohydrates, and 72 g of protein. The total energy value of this pizza is 1230Kcal. The pizza was divided into 6 pieces and participants had to consume each portion in 5 min during a total period of 30 min. Previous published and unpublished data (pilot study) have demonstrated that this fat overload model efficiently reduces the endothelium-dependent function of the brachial artery .After 10 min of quiet resting, a 30-min intervention to induce DBF or control) was conducted at three time points . Then, a pneumatic cuff (Hokanson) was placed around the left forearm and inflated to 75 mm Hg, and a second proximal cuff was positioned 3 cm distal from the axilla and inflated to 40 mm Hg to partially occlude venous flow from the arm. For normal flow measures, the cuff was placed on the same arm, but it was not inflated. Mean SR and the pattern of SR  were recorded by 60 s at three different time points, 0 (before the inflation of the cuffs), and 15 and 30 min (during inflation of the cuffs).Whole blood was sampled from the participants in EDTA tubes and then centrifuged at 448 relative centrifugal force (rcp) for 10 min at 4°C. Plasma was removed and kept aside for further analysis. The supernatant was stored at −80°C for subsequent assessments. Proteins were quantified by the method described by Nicotinamide adenine dinucleotide phosphate oxidase (NADPH) oxidase was determined by the rate of NADPH consumption assessed by measuring the decline in absorbance (340 nm) every 10 min, using a plate reader spectrophotometer  . For the2O2) was analyzed by measuring oxidation of phenol red  mediated by radish peroxidase , using a plate reader spectrophotometer  (Hydrogen Peroxide (HDevices) .Plasma lipid peroxide levels were determined by measuring thiobarbituric acid reactive substances (TBARSs), a common method for measuring the concentration of malondialdehyde, the main breakdown product of oxidized lipids. For the TBARS assay, using 250 μL of sample, trichloroacetic acid  was added to the homogenate to precipitate proteins and to acidify the samples. This mixture was then centrifuged , the protein-free sample was extracted, and thiobarbituric acid  was added to the reaction medium. The tubes were placed in a water bath (100°C) for 30 min. The absorbances were measured at 535 nm using a spectrophotometer  .Total antioxidant activity was determined using the ferric reducing/antioxidant power (FRAP) assay . Sodium +/CD42b– were determined by flow cytometry , as previously described  and during  experimental and control sessions. The endothelial microparticle populations CD31escribed . Briefly–1⋅). But the FMD was normalized by SR-AUC until peak vasodilation (peak diameter). FMD reliability was evaluated through the comparison of FMD value (pre-intervention) between days. The intraclass correlation coefficient was 0.93 (p < 0.05), and the coefficient of variation was 4.5%.The video files were compatible with commercial automated edge-detection and wall-tracking software , which was used for offline analysis. The initial phase of the software analysis consisted of identifying regions of interest on the optimal portion of the brachial artery image and its blood velocity spectra. R-wave gaiting function was applied to continuously assess brachial artery diameter or blood velocity. Flow-mediated dilation was calculated as the percentage rise of this peak diameter from the preceding baseline diameter. After deflation of the cuff, the cumulative shear rate was determined by a 3-min period through the area under the curve  and artery diameter (in cm) . For calculations of antegrade and retrograde SR, antegrade and retrograde mean blood velocities were used, respectively. In addition, oscillatory SR index, a variable that estimates the magnitude of oscillation in the vascular bed was calculated as IRetrograde shearI/IAntegrade shearI + IRetrograde shearI. Oscillatory SR index values range from 0 to 0.5, with 0 corresponding to unidirectional shear throughout the cardiac cycle, and 0.5 representing pure oscillation with time-average shear equal to 0. Oscillatory SR was expressed in arbitrary units (a.u.). The FMD, oxidative stress and EMPs response to experimental sessions is presented as—10 min (baseline) and time point 110 min. The absolute changes (delta) were calculated by subtracting values at 110 min from baseline.Shear rate (SR) was calculated as four times the ratio between mean blood velocity . These results clearly show that our model, via ingestion of mozzarella pizza, was able to induce systemic fat load.The metabolic responses to fat load in both PPL and PPL plus DBF sessions are displayed in the p < 0.05, to both variables, p < 0.05, p > 0.05, As expected, retrograde SR and oscillatory index responses to forearm cuff inflation at the three time points  were greater in PPL plus DBF compared with PPL . Delta analysis showed that brachial artery FMD% was decreased to a greater extent (at 110min) with addition of DBF  relative to basal (-10 min) .Our data revealed that both PPL plus DBF and PPL plus DBF did not change the resting diameter, peak diameter, SR-AUC, and time to peak  and antioxidant substances (FRAP), and lipoperoxidation (TBARS). PPL and PPL plus DBF similarly increased NADPH oxidase  released into the circulation in response to injury, activation, and apoptosis , then, tn = 18), and all were healthy. This does not allow us to say whether our results can be inferred for other populations, especially those who are at increased cardiovascular risk . Second, we did not evaluate the endothelium-independent function due to the prolonged effect of nitroglycerin on the vascular system and that repeated measures of endothelium-independent dilatation could compromise the experimental protocol. However, previous evidence has documented that acute metabolic changes or flow profile disturbances do not affect vascular smooth muscle reactivity in healthy subjects  associated with DBF has an acute additive effect on the reduction of brachial artery endothelium-dependent function 110 min after stimuli in healthy individuals, despite a similar increase in NADPH oxidase and hydrogen peroxide. PPL plus DPF and PPL did not modify plasma levels of TBARS and antioxidant defense, assessed by FRAP. Finally, PPL plus DBF and PPL similarly increased circulating EMVs. Further studies are needed to understand the mechanisms associated with endothelial dysfunction induced by the association of fat overload and blood flow disturbances and to propose preventive strategies  against these vascular insults.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by the Research Committee of the Heart Institute of Clinics Hospital of the University of São Paulo Medical School (CAAE 71981517.8.0000.0068). The patients/participants provided their written informed consent to participate in this study.GA and TS conceived experimental designs, performed experiments, interpreted results, and drafted the manuscript. HR and NR conceived experimental designs, interpreted results, and edited the manuscript. DF, JI, AD, and AA performed experiments and edited the manuscript. FMC-C and KA interpreted results and reviewed and edited the manuscript. MI and AS conceived and designed the study, interpreted results, and reviewed and edited the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


MiTNM staging was performed in both studies by two nuclear medicine physicians who were blinded to the results of the other tracer. After intradisciplinary and interdisciplinary consensus with two radiologists was reached, differences in both malignant and presumable nonspecific tracer accumulation were analyzed.A retrospective analysis of 46 prostate cancer patients (median age: 71 years) who underwent consecutive [18F]-F-PSMA-1007 PET. Concordant miTNM stages were obtained in 37 patients (80.4%). There was no significant difference between [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11 in the SUVmax locally , in lymph node metastases  or in bone metastases . In [18F]-F-PSMA-1007 PET, more patients featured presumable unspecific uptake in the lymph nodes , bones  and ganglia . Probable unspecific, exclusively [18F]-F-PSMA-1007-positive lesions mainly occurred in the ribs (58.7%), axillary lymph nodes (39.1%) and cervical ganglia (28.3%).Differences in terms of miTNM stages in both studies occurred in nine of the 46 patients (19.6%). The miT stages differed in five patients (10.9%), the miN stages differed in three patients (6.5%), and different miM stages occurred only in one patient who was upstaged in [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11, it appears reasonable to choose the PSMA radiotracer depending on local availability with attention to the greater occurrence of nonspecific bone findings with [18F]-F-PSMA-1007.In terms of miTNM staging, both tracers appeared widely exchangeable, as no tracer relevantly outperformed the other. The differences between the two tracers were far more common in presumable unspecific lesions than in malignant spots. A routinely performed two-tracer study could not be shown to be superior. Since it seems at least challenging for most nuclear medicine departments to provide both [ Within nitoring –9.68Ga]-Ga-labeled radiotracers, the capacity of the examination of [68Ga]-Ga-PSMA-11 is limited by its short half-life of only 68min, requiring in most cases in-house production and a sufficient generator supply. However, cyclotron-based production methods have recently been developed [68Ga]-Ga-labeled tracers equivalently to [18F]-F-tracers. Nevertheless, the commercial availability of [68Ga]-Ga/[68Ge]-Ga-generators allows cyclotron independent tracer production for institutions, that have no such access.However, as in all [eveloped . The out18F]-F-PSMA ligands may overcome the limitations of the short half-life of [68Ga]-Ga. Furthermore, the end point positron energy of [18F]-F is much lower than that of [68Ga]-Ga (0.65 vs. 1.90 MeV), which reduces the positron range in tissue and may improve spatial resolution [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11.The implementation of [solution . The dif18F]-F-PSMA-1007 features a lower urinary excretion than [68Ga]-Ga-PSMA-11 -Ga- and [18F]-F-PSMA-ligands in general [18F]-F-PSMA-1007 [68Ga]-Ga-PSMA-11 specifically are considered widely exchangeable for most indications. However, there are only a few clinical studies directly comparing [68Ga]-Ga-PSMA-11 and [18F]-F-PSMA-1007. Matched comparisons in patients with biochemical recurrence (BCR) [18F]-F-PSMA-1007 has indicated more nonspecific lesions -Ga-PSMA-11 and [18F]-F-PSMA-1007 PET/CT or PET/MRI. Two patients received therapeutic measures between the two studies and were therefore not eligible for analysis. Seven more patients were excluded due to a lack of clinical data, no interdisciplinary tumor board presentation or missing consent for scientific re-evaluation . All other patients received [68Ga]-Ga-PSMA-11 PET first. All 46 patients in this retrospective single-center analysis were discussed by an interdisciplinary tumor board, and an interdisciplinary consensus of all imaging results and further therapeutic management were decided upon. Written informed consent was obtained from all patients for the clinically indicated examination and the consecutive scientific analysis of their clinical and imaging data. The institutional review board of the local ethics committee at our medical faculty approved this study.Between 07/20 and 12/20, fifty-five prostate cancer patients underwent both [68Ga]-Ga-PSMA-11 [18F]-F-PSMA-1007 -Ga-PSMA-11 examinations, a median of 149 MBq (range: 111–161 MBq) was intravenously injected, and acquisition started after a median of 106 min p.i. , while for [18F]-F-PSMA-1007 PET, a nearly equal median of 154 MBq (range: 123–175 MBq) was applied, and imaging started after a median of 103 min p.i. .For the [18F]-F-PSMA-1007 and 1.4 mm/s for [68Ga]-Ga-PSMA scans.The PET/CT scans of 40 patients were acquired using a Biograph Vision 600 device . A low-dose CT scan was acquired from the whole body  and used for attenuation correction of the following PET scan. The emission PET scan was obtained using continuous bed motion with a speed of 2.2 mm/s for -Ga-PSMA-11 studies and the spleen in studies with biliary excreted [18F]-F-PSMA-1007. If a lesion was not separable from the local background at one time point, it was scored as 0, regardless of its SUVmean. In this case, it was excluded from any further analysis. Sufficiently large -Ga-PSMA-11 and [18F]-F-PSMA-1007. The median PSA value of all patients was 3.76 ng/ml (range: 0.3–113.7 ng/ml).A total of 46 prostate cancer patients at a median age of 71 ± 8.0 years and a median disease duration of 41 months (range: 0–192) underwent analysis with both -Ga-PSMA-11. There were no differences in blood pool uptake . The [18F]-F-PSMA-1007 uptake did not differ significantly between the liver and spleen (p = 0.615). However, the relevant reference tissue uptake for the miPSMA score was higher in the spleen for [18F]-F-PSMA-1007 than in the liver for [68Ga]-Ga-PSMA-11 (p < 0.001).[18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11 PET were evaluated independently in terms of miTNM staging (Table Both -F-PSMA-1007 PET, while in [68Ga]-Ga-PSMA-11 PET, there was additional peripheral uptake in the right peripheral lobe  and ADT were initiated . The SUVpeak of [18F]-F-PSMA-1007, however, was only higher in eight patients, while the SUVmean did not differ significantly (p = 0.961). The miPSMA score, with different reference tissues in both tracers, was identical in 13 patients; only three patients had a higher prostatic miPSMA score in [18F]-F-PSMA-1007, while six had a lower miPSMA score.In the 22 patients with tracer uptake in the prostate or in the prostatic bed in at least one study, the SUVmax  and SUVpeak  of the main lesions did not differ significantly between [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11.The median SUVThe MiN0 stage was concordantly indicated by both tracers in all 32 patients (69.6%) Table .Table 5m18F]-F-PSMA-1007 PET and in a little lesser 28 regions  in [68Ga]-Ga-PSMA-11 PET. [18F]-F-PSMA-1007 indicated more lesions than [68Ga]-Ga-PSMA-11 in five regions , while the opposite was the case only in one internal iliac region . Eighteen of 46 (39.1%) patients had pelvic and/or extrapelvic lymph node metastases. The 14 patients with pelvic lymph node metastases had these in a total of 31 pelvic regions in [18F]-F-PSMA-1007 PET led to upstaging from miN1 to miN2, while downstaging was not observed.Differences in terms of miN staging were obtained in only three patients (6.5%) and only between miN1 and miN2 . Other, mostly additional locations were rare: supraclavicular (n = 1), hilar (n = 2), mediastinal (n = 1), axillary (n = 1) and inguinal (n = 1). In addition to his concordant hilar and mediastinal lymph node metastases and pulmonary metastasis, one patient had solitary paraureteral iliac lymph node metastasis solely detected by [18F]-F-PSMA-1007  in both [1007 and 8Ga-Ga-PS.4% in bomax  and SUVpeak  of the most intense lymph node metastases did not differ significantly between [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11.The median SUVThe different tracer distributions between the two tracers in terms of distant metastases led to a different miM stage in only one patient Table .18F]-F-PSMA-1007 PET , miM1a , miM1b  or miM1c .The remaining patients were concordantly staged as miM0  showed unifocal (n = 6), oligofocal (n = 3) or multifocal (n = 3) uptake in osseous metastases.Bone metastases were the most frequent manifestation of miM1. In [68Ga]-Ga-PSMA-11 PET, 11 patients (23.9%) showed unifocal (n = 7), oligofocal (n = 1) or multifocal (n = 3) uptake in osseous metastases. No patient had exclusive bone metastases on [68Ga]-Ga-PSMA-11 PET. Within the three patients with multifocal bone metastases, only concordant metastases occurred. Table In -F-PSMA-1007 and [68Ga]-Ga-PSMA-11.The median SUVn = 2) and soft tissue metastases (n = 1) were rare and were concordantly indicated in both studies. Figure Pulmonary .Most likely, unspecific, discrete (miPSMA score 1) lymphonodal uptake was present in 24 (52.2%) of the [18F]-F-PSMA-1007 indicated  exclusive presumable unspecific lymphonodal foci in 18 different patients (39.1%). While additional axillary lymphonodal uptake was noted in all 18 patients (39.1%), 6 patients had unspecific mediastinal/hilar uptake (13.0%), and 5 patients presented with unspecific auxiliary inguinal tracer accumulation. A patient with axillary lymph node uptake that was interpreted as unspecific is exemplarily shown in Fig. 68Ga]-Ga-PSMA-11.[atients 3.1%. Whil68Ga]-Ga-PSMA-11 PET, with 11 patients (23.9%) presenting unifocal (10.9%) or oligofocal (13.0%) bone uptake without implication of malignancy. Neither multifocal presumable unspecific bone uptake nor exclusive lesions were present.Osseous uptake not suggestive of malignancy was quite rare in [18F]-F-PSMA-1007 PET, a large majority  of the patients featured a low-intensity (miPSMA score 1) bone uptake, which was unifocal (8.7%), oligofocal (15.2%) or mostly multifocal (47.8%) and unsuspicious for malignant origin.In [18F]-F-PSMA-1007 PET indicated exclusive osseous lesions in 28 patients (60.9%). Twenty-seven of those patients had additional uptake in the ribs . As shown in Table 18F]-F-PSMA-1007 were the spine (23.9%), pelvis (10.9%) and scapula (10.9%). Figure 18F]-F-PSMA-1007 PET that were interpreted as unspecific.1F-F-PSMA-ients 60.%. Twenty18F]-F-PSMA-1007 studies (71.8% vs. 43.5%). The cervical ganglia were affected in all 13 cases (28.3%) with additional ganglionic uptake in [18F]-F-PSMA-1007 PET. Exclusive tracer uptake in the coeliac (19.6%) and presacral ganglia (10.9%) was detected less frequently and always in addition to cervical ganglia uptake.Small focal uptake in the cervical, coeliac and sacral ganglia was present more often in the [68Ga]-Ga-PSMA-11 PET than in [18F]-F-PSMA-1007 PET , while uptake in the thyroid gland occurred almost exclusively with [18F]-F-PSMA-1007 .Homogeneous esophageal uptake that was not suspicious for malignancy was more common in -F-PSMA-1007 PET, a device-dependent difference in presumable unspecific uptakes was observed between PET/CT and PET/MRI. Non-malignancy-associated uptake in bones and ganglia  occurred more frequently or even exclusively, as in lymph nodes , on PET/CT. As shown in Table 68Ga]-Ga-PSMA-11 PET.In [68Ga]-Ga-PSMA-11 and [18F]-F-PSMA-1007 led to disconcordant tracer accumulations, especially in lesions of unspecific/benign origin and to a far lesser extent in prostate cancer manifestations, which is in line with the previously published head-to-head [The different biodistributions of [-to-head  and matc-to-head .18F]-F-PSMA-1007 may lead to superiority against [68Ga]-Ga-PSMA-11 [The far lower urinary tracer excretion of [-PSMA-11  and rena-PSMA-11  in terms18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11 in terms of identifying intermediate- and high-risk prostate cancer manifestations in primary staging, with [18F]-F-PSMA-1007 indicating additional low-grade lesions. In accordance, miT staging was based on concordant lesions in nearly 90% of patients. Histologically confirmed exclusive lesions (after RPx or biopsy) were detected for each tracer once, implicating that there is no systematic advantage of one tracer in terms of primary staging. No significant difference in terms of SUVmax or SUVpeak was noted.Kuten et al.  showed a18F]-F-PSMA-1007 might be superior in terms of recurrence detection [68Ga]-Ga-PSMA-11 could be mitigated by diuretic premedication [68Ga]-Ga-PSMA-11. An explanation therefore might be found either in the different PSMA affinities or, ironically, in the lower spatial resolution of [68Ga]-Ga-PSMA-11. Lower resolution might be a reason for two or more small adjacent tumor nests to be recognized as one focus in [68Ga]-Ga-PSMA-11, while they are obscured in [18F]-F-PSMA-1007 as simple heterogeneity.Even though not relevantly advantageous in terms of general miTNM staging, -F-PSMA-1007 [68Ga]-Ga-PSMA-11 PET and the spleen in [18F]-F-PSMA-1007 PET were significantly different, no further lesion-based comparisons were performed based on scoring. Intrapelvic lymph node metastases were concordantly excluded in nearly 70% of all patients (N0 situation).Differences in the detectability of lymph node metastases may occur both because of different total uptake of the lesion and because of altered levels of the local tumor background, which may be higher in the pelvis in biliary excreted [SMA-1007 . As the 18F]-F-PSMA-1007 PET compared with [68Ga]-Ga-PSMA-11 PET, while the opposite was the case in just one site. An explicit region-based analysis could not identify a predilection spot of superiority. In addition to the not explicitly evaluated tumor-background ratio (TBR), a potential locoregional diagnostic advantage of biliary excreted [18F]-F-PSMA-1007 could not be shown, as both SUVmax and SUVpeak were not significantly different. Patient-based differences were even rarer. In [18F]-F-PSMA-1007 PET of three patients, the number of infested lymph node regions increased, leading to an upstaging from N1 to N2. From a clinical point of view, these differences did not influence further therapeutic management.Thus, differences in miN stages occurred only in the number of metastases within one region or the number of regions harboring lymph node metastases. A region-based evaluation of the 14 patients with concordant miN1 or miN2 revealed only 5 additionally infested lymph node regions in [18F]-F-PSMA-1007 indicated more-predominant or exclusive bone metastases in one patient each. Thus, only in one patient who had BCR and concordantly indicated pelvic lymph node metastasis did [18F]-F-PSMA-1007 PET indicate a higher miM stage. No tracer outperformed the other in terms of SUVmax and SUVpeak. The few detected PSMA-positive pulmonary and soft tissue metastases were focally consistently indicated by both tracers.For extrapelvic lymph node manifestations, all lesions were indicated concordantly by both tracers. The miM staging was shown to be even more concordant (97.8%) between the two tracers than the miT and miN staging. [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11 can be considered exchangeable -F-PSMA-1007 PET -F-PSMA-1007 PET. This pattern is well known -Ga-based [18F]-F-based ligands -Ga-PSMA-11 [18F]-F-PSMA-1007. One possible explanation of the higher SUVmean in the liver, spleen and parotid glands evaluated with [18F]-F-PSMA-1007 compared to [68Ga]-Ga-PSMA-11 is the higher spatial resolution of 18F compared with 68Ga as a result of the lower end-point positron energy. This may have caused a more focal impression in [18F]-F-PSMA-1007 images. We will further investigate in detail the individual influence of the physical properties of the nuclide and the reconstruction process on SUV values, image quality and spatial resolution.The high rate of tracer accumulation attributed to benign genesis is already known for -F-PSMA-1007, and homogenous esophageal uptake was more common with [68Ga]-Ga-PSMA-11.In addition to these measurement factors and biokinetic aspects, another explanation focuses on the PSMA ligand itself, which has been shown to have different affinities for different tissues , 43, 44,18F]-F-PSMA-1007. As clinical stages progress, the advantages of [18F]-F-PSMA-1007 over [68Ga]-Ga-PSMA-11 possibly fade, as the exact identification of all intrapelvic regions loses increasing clinical and therapeutic relevance. In terms of the evaluation of PRLT in Stage IV patients, [68Ga]-Ga-PSMA-11 is possibly even superior, as its biodistribution is closer to that of [177Lu]-Ga-PSMA-617 than to that of [18F]-F-PSMA-1007 [68Ga]-Ga-PSMA-11 in PRLT evaluation, however, will rarely yield clinical consequences, as both tracers will mostly consensually lead to an indication for PRLT, regardless of the exact number of PSMA-positive lesions. In the oligometastatic setting, these differences are of higher importance, as exclusively indicated lymphonodal or osseous tumor manifestations have therapeutic implications -Ga-PSMA-11 -F-PSMA-1007 and [68Ga]-Ga-PSMA-11 were, regardless of the indication, widely exchangeable in terms of metabolic TNM staging of prostate cancer.In our retrospective head-to-head comparison, the radiotracers [18F]-F-PSMA-1007 indicates more presumable unspecific lesions than [68Ga]-Ga-PSMA-11 but is advantageous in terms of tumor delineation. A significant incremental value for a routinely performed two-tracer study could not be shown. As it seems for most nuclear medicine departments to be at least challenging to provide both [18F]-F-PSMA-1007 and [68Ga]-Ga-PSMA-11 PET, it is reasonable to choose the PSMA radiotracer depending on local availability with attention to the greater occurrence of nonspecific bone findings with [18F]-F-PSMA-1007.However, there are assumptions that [