Parkinson’s disease (PD) is a neurodegenerative disease and a multidisciplinary approach to rehabilitation has been suggested as the best clinical practice. However, very few studies have investigated the long-term effects of a multidisciplinary rehabilitation approach, particularly regarding whether this can slow the progression of PD. The purpose was to investigate the short- and long-term effect of a 2-week multidisciplinary rehabilitation regimen on the PD-related decline in health-related quality of life (HRQOL), mobility, and muscle function. Individuals with PD (IPD) participated in a 2-week inpatient multidisciplinary rehabilitation regimen that focused on improving HRQOL, mobility, and muscle function. Data from the primary outcome: HRQOL , secondary outcomes: handgrip strength, Timed-up and Go (TUG), Hospital Anxiety and Depression Scale (HADS), and Falls Efficacy Scale-International (FES-I) were compared at pre-visitation, before and after the 2-week regimen, and again at 4 and 10 months follow-up. In total, 224 patients with PD were included. There were short-term improvements in all outcomes. PDQ-39 was maintained at the same level as pre-visitation after 10 months follow-up. A 2-week multidisciplinary rehabilitation regimen improved short-term mobility, muscle function, and HRQOL in individuals with Parkinson’s disease. HRQOL was maintained after 10 months demonstrating long-term effects. Parkinson’s disease (PD) is a progressive neurodegenerative disease caused by a deficiency in the neurotransmitter dopamine in basal ganglia . PD affeIt is well-documented that reduced muscle strength in healthy elderly is linked to functional deficits such as impaired walking speed and transfer  and thatAs PD is a disease affecting multiple aspects of life, international PD rehabilitation guidelines  recommenTo address this limited knowledge, the objective of the present feasibility study was to investigate the effect of a 2-week inpatient multidisciplinary rehabilitation regimen with long-term follow-up at 10 months that focused on improving HRQOL, mobility, and muscle function in IPD. In addition, the study sought to collect data from a short-period of a multidisciplinary rehabilitation regimen primarily consisting of patient education to improve physical and mental health  for IPD and documents the programs feasibility for future planning of a larger RCT. Data were collected at pre-visitation, before and after the regimen, and at 4 and 10 months follow-up. We hypothesized that IPD would benefit from a 2-week inpatient multidisciplinary rehabilitation regimen by improving mobility, physical function, and health-related quality of life HRQOL with lasting effects.This study is designed as a prospective cohort feasibility study to assess if a short-term regimen of 2-weeks could have a positive impact on HRQOL. IPD were recruited from a specialized PD outpatient clinic at the Department of Neurology, Bispebjerg University Hospital, Copenhagen, Denmark. When recruited, the IPD’s neurological status was assessed, including the severity of PD as evaluated on Hoehn and Yahr (H and Y) stages of disease 1–5  . SubsequIPD stayed at the center for 2 weeks, this in order to minimizes logistical problems with transportation and optimize the intensity of the regimen. Inclusion- and exclusion criteria are presented in All participants provided written informed consent before initiation of any study procedures, and the study was complied with the Helsinki Declaration.The multidisciplinary team consisted of a neurologist, neuropsychologist, psychologist, sex therapist, speech therapist, physiotherapists, occupational therapists, and a nurse. All team members had expert knowledge in the treatment of PD.The 2-week multidisciplinary rehabilitation program was delivered sequentially to groups of 8 participants. The main components of the program were (a) a comprehensive introduction to resistance-training and (b) daily training supervised by physiotherapists with the purpose of introducing and testing alternative activities with physical and psychological elements. IPD’s were introduced to Nordic Walking, aqua training in a heated pool, meditation, and dancing.Furthermore, health promotion initiatives and lectures with a wide variety of relevant issues for IPD were given. A neurologist held a lecture on PD, updated on the newest evidence with concern of treatment, typical symptoms, and prognosis. A neuropsychologist had focus on insight and understanding of the stress load that accompanies the disease; tools were given to handle stress and reduce the impact on daily living. The session contained theoretical presentation and exercises which involved participants. A psychologist gave insight in the emotional reactions with PD, coping with stress, griefs, and crisis. The psychologist also held a special session for the relatives in order for them to gain more knowledge on PD in a closed environment, where they also were able to exchange experiences. Nutrition was reviewed by a nurse with focus on the composition of a nutritious diet. Occupational therapists gave information on assistive devices. Furthermore, they held sessions on coping with tools to discover their own resources and implementing them in daily living. A sex therapist informed on sexuality and cohabitation with advice and guidance on the possibilities for functioning as well as possible challenges that may arise when one party is affected by PD. Lastly, a speech therapist presented knowledge on voice strength and how it might be affected by PD, respiration and communication exercises see . RelativAll IPD were given an in-depth introduction to a simple resistance-training program targeting balance, muscle strength, and function. The duration of a single training session was 30–45 min. The program was suitable for training either in the local outpatient physiotherapy clinic, gym, or participants’ homes. The program was designed according to the American College of Sports Medicine guidelines to obtain the best effect and compliance .The resistance-training program was provided/delivered by an experienced physiotherapist three times per week for a total period of 2 weeks . Each training session began with 10-min warm-up exercises of light to moderate intensity on a stationary bike followed by the resistance-training program, which consisted of 3−5 bilateral exercises such as leg press, leg curl, leg extensions push, and pulldown. The participants completed 2−3 sets of each exercise with 1–2-min rest between each set. The repetitions/loads were: Week 1: 12 repetitions with loads corresponding to 15 repetition of maximum (RM). Week 2: 12RM-9RM . PatientAll participants received a logbook. To improve compliance for continued training, the participants’ local physiotherapist was informed about the rationale including the evidence behind the PD resistance-training program. Participants received two strength training programs: one program based on training on machines at the gym, and one home-based that could be implemented without machines and equipment such as backpack, bags with water containers and TheraBand (elastic training band). Both programs could be used by the local physiotherapist and were not dependent on advanced training equipment.In order to accommodate any questions concerning training, participants had the opportunity to get into contact with the study physiotherapist by phone on a weekly basis during the four months follow-up. Furthermore, participants were contacted by a physiotherapist by phone at 5 months and 7.5 months after completion of the multidisciplinary rehabilitation program in order to improve adherence to continued training .Physical and HRQOL assessments were performed at the following timepoints: pre-visitation, start-rehabilitation regimen, end-rehabilitation regimen, and 4 and 10 months after ended intervention see .All tests were conducted in the morning and approximately 1 h after medication intake; physical tests were performed at the rehabilitation centers by therapists who were not involved in intervention delivery. Questionnaires were collected by administrative personnel with no other role in the study. All assessors and participants were unaware of earlier assessment results.Primary Outcome: HRQOL using Parkinson’s disease questionnaire 39 (PDQ-39) . PDQ-39 Handgrip strength. This assessment was first performed on “the most affected side” and then on “the least affected side”, which contained three trials with a short pause 20–30 s) between each attempt. The mean value of the three trials was calculated and entered in the statistical analysis. A North Coast Digital Hand Dynamometer  was used [ s betweet-test). The correlation between first and second measurement was 0.95. No clinically important differences in TUG has been determined for IPD; however, the minimal detectable change  values range from 3.5 to 11 s [Timed-Up-and-Go (TUG) was used to test gait function and balance. Participants were timed as they rise from a chair, walk 3 m, turn, and return to sitting on the chair; assistive devices were allowed if needed . High va to 11 s .Hospital Anxiety and Depression Scale (HADS): HADS was administrated by neuropsychologists and used as a screening tool for the identification of anxiety and depression. HADS contains 14 questions: seven questions to assess anxiety and seven to assess depression . High scFalls Efficacy Scale-International (FES-I): Self-reported questionnaire with 16 questions concerning fear of falling. Item scores range from 1 (no worries) to 4 (very worried) . Total sSecondary outcomes:p < 0.01. SAS version 9.4  was used for the statistical analysis.Means, standard deviations (SD), and number of available observations are shown for each of the outcomes at each of the five assessment time points. Raw effect sizes are calculated for the change from pre-visitation, start-rehabilitation regimen to end-rehabilitation regimen, 4 months and 10 months, respectively. A multilevel linear regression model was used to assess adjusted mean differences in outcomes between the different time points. Comparisons were made between the following time points: pre-visitation vs. start-rehabilitation regimen, end-rehabilitation regimen, 4 months and 10 months; start-rehabilitation regimen vs. end-rehabilitation regimen, 4 months and 10 months, end-rehabilitation regimen vs. 4 months and 10 months. The model accounted for the inherent excess correlation between the up to five measurements per patient in the study through a patient random effect. The models were adjusted for potential confounding by gender, age and PD severity (H and Y stage). Data were used from all IPD’s, including those who dropped out. Potential differential dropout was adequately handled with the use of the multilevel model . The sigWe recruited 224 IDP’s between June 2011 and June 2014, of whom 214 IPD completed the 2-week multidisciplinary rehabilitation program. In total, 112 IPD were treated at VRC and 112 IPD were treated in CTS. A total of 10 drop-outs were encountered. See Participant characteristics stratified by rehabilitation center are described in p < 0.001). At 10 months PDQ-39 score increased (p < 0.001) to the same level as observed at pre-visitation.As illustrated in p < 0.001) during the 2-weeks regimen. At 4 months, grip strength was improved by 10% (effect size: 0.3) on “the most affected side” from 30 to 33 kg (p < 0.001) and 3% (effect size: 0.1) on “the less affected side” from 34 to 35 kg (p < 0.001).Handgrip strength increased by 7% (effect size: 0.2) from 30 to 32 kg (most affected side), (p < 0.001). After 4 months, an improvement by 17% (effect size: 0.4) from 8.5 to 7.1 s (p < 0.001) was observed.TUG reduced by 14% (effect size: 0.4) during the 2-week regimen from 8.5 to 7.3 s (p < 0.001). At 4 months, the depression score was almost back to the level before the multidisciplinary rehabilitation (p < 0.01). No other differences were found. Results are presented in The HADS score, reduced by 18% (effect size: 0.3) for depression and anxiety during the 2-week program (p = 0.016).For FES-I, no significant differences were found at any time points, but a trend towards a reduced score was observed between pre-visitation and 4 months (The main findings of this feasibility study were that patients with PD (phase 2–3) improved HRQOL (measured by PDQ-39), mobility, and muscle function (measured by TUG) and handgrip strength on both sides after completion of a 2-week inpatient multidisciplinary rehabilitation program. Furthermore, at 10 months follow-up HRQOL (PDQ-39) was maintained at the same level as at pre-visitation demonstrating long-term effects. These findings suggest that a 2-week inpatient multidisciplinary rehabilitation approach consisting of IPD education with focus on activities that promote HRQOL and physical activity exercise such as using a simple home-based resistance-training program is likely to improve function and sustain long-term HRQOL.Recently, a systematic review by Johnston and Chu  showed tHandgrip strength has been described as one of most simple methods for assessing general muscle strength and function . In our TUG is a well-proven test and recommended in the Dutch clinical guidelines for PD . In the The HADS score demonstrated a reduction for depression and anxiety after the 2-week multidisciplinary rehabilitation; however, this was not clinically significant as Minimal Clinically Important Difference (MCID) should be beyond 1.3 (depression) and 1.4 (anxiety). Interestingly, the depression component increased level at 4 months was almost at same level as pre-visitation, however this did not apply to the anxiety component. HADS is designed for screening mood disorders in general (non-psychiatric) medical outpatients and aims at distinguishing depression from anxiety . One canp = 0.016) was demonstrated, indicating that patients had less fear of falling. Together, this indicates that progressive strength training could influence patients positively in the fear of falling.A previous study by Goodwin , has demOur study has several limitations. One of them is the lack of a control group or control intervention and it is, therefore, unknown whether a placebo effect was responsible for the greater results. The present prospective cohort feasibility study design was chosen to assess the feasibility of the intervention, relevance of selected outcome measurements and possible effects before performing a larger randomized controlled trial. Therefore, pre-visitation was held 3 months before the 2-week inpatient multidisciplinary rehabilitation period and with long-term follow-up at 4 and 10 months. Furthermore, only PDQ-39 was assessed at 10 months follow-up, but it could also have been interesting to assess outcomes such as functional parameters  and socioeconomic aspects including comorbidities. However, this was not feasible in the present study. Future studies could include these components, but also dosage and intensity of physical training need to be further investigated in order to tailor a sufficient treatment program for the individual patient. Finally, future studies should address the economic benefits with only the 2-week inpatient multidisciplinary rehabilitation regimen and assess the potential cost savings in terms of prevented hospitalization due to falls and injury.A 2-week inpatient multidisciplinary rehabilitation program consisting of simple progressive physical exercises, cognitive behavioral therapy, patient education (including Parkinson management and lifestyle changes), and a socially supportive environment improved health-related quality of life (PDQ-39), mobility and muscle function. Health-related quality of life was maintained after 10 months demonstrating long-term effects.


All three α1-AR subtypes  are expressed in human and animal hearts and blood vessels in a tissue-dependent manner. We summarize the changes observed in HF regarding the density, signaling and responses of α1-ARs. Conflicting findings arise from different studies concerning the influence that HF has on α1-AR expression and function; in contrast to β-ARs there is no consistent evidence for down-regulation or desensitization of cardiac or vascular α1-ARs. Whether α1-ARs are a therapeutic target in HF remains a matter of debate.As heart failure (HF) is a devastating health problem worldwide, a better understanding and the development of more effective therapeutic approaches are required. HF is characterized by sympathetic system activation which stimulates α- and β-adrenoceptors (ARs). The exposure of the cardiovascular system to the increased locally released and circulating levels of catecholamines leads to a well-described downregulation and desensitization of β-ARs. However, information on the role of α-AR is limited. We have performed a systematic literature review examining the role of both cardiac and vascular α In contrast, antibody-based approaches such as immunoblotting or immunohistochemistry potentially can be subtype-selective and be used to examine distribution within a tissue. However, multiple studies have demonstrated that the vast majority of commercially available α1-AR subtype antibodies lack target specificity when tested under stringent conditions [Cardiac αnditions ,21,22. T1-ARs in the peripheral vasculature. It has been demonstrated that α1-AR receptors are located not only on smooth muscle cells, but, also on endothelial, adventitial and nerve cells [1-AR mRNA and the actual role of specific α1-subtypes mediating vascular responses might differ depending on the specific cell on which they are expressed.Another important factor that might influence the results of different studies is the exact location of αve cells ,24. This1-ARs. Finally, pathophysiology and regulation of α1-ARs may not only differ between patients and experimental models but also between species. While most animal studies related to cardiovascular α1-ARs in CHF were done with rats [CHF can be caused by different pathologies, for instance by chronic myocardial ischemia, valvular heart disease or viruses, or can be idiopathic. As CHF in humans can be caused by a variety of pathophysiologic mechanisms, it is important to realize that subjects within an animal model typically are more homogeneous than patient cohorts; on the other hand, each model reflects a different cause and, potentially, a different pathophysiology. Among the many reported animal models of CHF, post-myocardial infarction ,26,27, cith rats , guinea ith rats ,34, miceith rats , hamsterith rats , cats [3ith rats  and dogsith rats ,38 have 1-AR. One study was reviewed for both cardiac and vascular α1-ARs leading to a total number of included studies of 65.The literature search resulted in 1461 unique, original articles. G.K.-M. and O.P. independently screened the titles and abstracts of all the articles and excluded 1312 articles because of lack of relevance. After assessing the full text of the remaining 149 potentially relevant articles, and resolving any disagreements, the two reviewers decided to include 65 articles . Additio1-AR subtypes: α1A-, α1B- and α1D-ARs which have been identified in both animal and human hearts at the mRNA level [1A-AR is the most abundant subtype in the heart but the presence of α1B- and α1D-AR mRNA were also confirmed [1-AR in the heart, human and rodent hearts are reported to locate α1-AR subtypes similarly, as α1A- and α1B-subtypes are predominantly found in myocardium whereas the α1D-subtype is expressed in coronary arteries [1A-ARs are present in a myocyte subset while α1B-ARs are in all cells [1-AR subtype mRNA levels in CHF, but the results are heterogeneous  suggesting that this subtype may contribute to maintain cardiac inotropy in the failing heart. Moreover, a study in a rat coronary ligation model of CHF reported α1D-AR mRNA to be decreased; a numerical reduction of α1A- and α1B-AR mRNA did not reach statistical significance [In human failing hearts, Fischer et al. demonstrated that mRNA of α regions . Similary region . On the y region . They suificance .1-ARs were first identified at the protein level in rats by Williams and Lefkowitz [1-AR binding properties in human HF have remained inconsistent , which releases Ca2+ from intracellular stores and activates protein kinase C (PKC) [1-ARs can also couple to Gi [12/13 [s proteins [h which is a GTP-binding protein with Transglutaminase activity [2+, K+ and Cl- channel activity [A schematic representation of α C (PKC) . Howeverle to Gi ,7, G12/1i [12/13 , even Gsproteins  and Gh wactivity  and to pactivity , modulatactivity ,69, ras activity  or activactivity . Most ofactivity .q-signaling pathways mediates the development of hypertrophy and the transition to HF [q was demonstrated to result in marked hypertrophy [q-mediated signaling was shown to inhibit cardiac dysfunction [i at the mRNA and protein level, whereas a reduced expression of Gs has been found less consistently [h were found in human HF [1-AR coupling to a GTP binding protein was detected only in pathological, but not in healthy human hearts [Activation of Gon to HF . Overexpertrophy  and HF [ertrophy . Moreovefunction ,77. On tistently . Reducedhuman HF . α1-AR cn hearts .1-AR-mediated phosphoinositol turnover is not changed in both animal and human studies when accompanied by an unaltered receptor density in human [1-ARs in the reactive hypertrophic response [αin human  and rat in human  HF. On tresponse .1-AR stimulation increases both the intracellular Ca2+ concentration, which activates muscle contraction, but also the Ca2+ sensitivity of cardiomyocytes. α1-AR-mediated Ca2+-transients and phosphorylation of cardiac myosin regulatory light chain were increased in a mouse right ventricular HF model [2+ sensitivity expected from these observations was reported in a canine HF model, apparently occurring via the Gq-RhoA-ROCK signaling pathway [2+ sensitivity [αHF model . The inc pathway . Similarsitivity .1-AR stimulation induces a positive inotropic effect in isolated cardiac preparations such as atria, left and right ventricles  [1-AR mRNA expression can be regulated by hypoxia [1-AR mRNA expression is also regulated by age [1-AR mRNA in human coronary arteries [1D and total α1-AR mRNA levels. α1D-AR appeared to be the principal receptor in human epicardial coronary arteries, which resembles the mouse coronary arteries [Multiple studies have explored the expression of αding rat ,86,87,88ding rat ,90,91 anding rat ,92,93,94st 96 h) . Moreoved by age . HF did arteries . On the arteries ,100.1-AR distribution in the various vascular beds of different species. Existing discrepancies might be due to interspecies differences, variant α1-AR subtype distribution in the peripheral vasculature, differences in models of HF, for example pacing-induced HF versus myocardial infarction, or even due to poorly reproducible results.Quantitative, ligand-binding studies have been performed in order to assess the α1-ARs at the protein level. In humans a considerable difference in α1-AR protein expression has been demonstrated between somatic (mammary artery) and splanchnic arteries [1-AR density seemed to be independent from differences in vessel diameter. This might be explained by the importance of splanchnic circulation, especially during a situation of hypovolemic shock. Sympathetic stimulation of α1-ARs in those vascular territories leads to maintenance of blood flow and O2 delivery to vital organs. On the other hand, constriction of conduit arteries and veins contribute to the preservation of blood pressure and cardiac afterload. In human epicardial coronary arteries 8.7 fmol/mg protein of total α1-ARs were found, of which 75% constituted of the α1D subtype, almost twice as much as in the left ventricle [Such studies have been performed in animals and humans without coexisting diseases in order to assess the physiological αarteries . Howeverentricle ,98.1-ARs. On the contrary, α1-AR density has been reported to be increased, yet not statistically significant, in studies with myocardial infarction-induced CHF in rats [Considering that progression of HF is associated with a decrease in β-AR protein in the heart, one could argue that similar findings would be expected for α in rats ,102. How1B and α1D subtype mRNAs have been detected in human aorta [1D [1B and α1D-ARs [1B subtype has been recognized to mediate vasoconstriction after sympathetic stimulation in the mouse, rabbit and dog aorta [1-AR subtypes [1D and α1B subtypes in superior mesenteric artery and the mesenteric resistance arteries respectively [1A has also been identified [All three subtypes have been identified in the peripheral vasculature of various species, however the subtype distribution varies between animals and humans, between different vascular beds and with age. For instance, αan aorta  and are orta [1D ,104,105, α1D-ARs . The α1Bog aorta ,107,108.subtypes , whereasectively ,104. In entified ,110. Theentified .1D is recognized as the main subtype that mediate coronary contraction in humans [1D-AR, neither in the carotid artery nor in the aorta, whereas the α1D-AR-mediated vasoconstriction was enhanced in older animals.Similarities in vascular tone regulation between different species also exist. As it was mentioned before, αn humans  and micen humans ,100 and n humans . In thisFurthermore, blood pressure regulation in mice is considered to be achieved via the stimulation of all three subtypes. This is confirmed in knockout mice studies in which deletion of any subtype results in a decrease in resting blood pressure ,112,113.1-AR population in the various vascular beds might be modified depending on the disease state. In support of this, antagonist studies have explicitly displayed an alteration of the antagonism against α1-ARs before and after the onset of CHF [The heterogeneous αt of CHF ,116,117 1-AR responsiveness due to chronic exposure of the peripheral vasculature to the endogenously released catecholamines might also derive from alterations in signal transduction. These changes might be due to modification of Ca2+ sensitivity and/or other changes in the signaling pathways involved. For example, the continuous exposure of smooth muscle cells from rabbit aorta to α1-AR agonists led to a major loss in sensitivity, which did not seem to be due to down-regulation of α1-ARs, but rather to blunting of phosphatidylinositol turnover [Changes in αturnover .1-AR-mediated vascular contraction on extracellular Ca2+ which seems to increase in end-stage HF [1-AR-mediated responses was identified which is inhibitable by a PKC inhibitor, polymyxin B, and which becomes apparent with the progression of HF. Interestingly, in this study, the dorsal pedal artery presented lower dependency on PKC activation than the saphenous vein, which underlines the existing differences between separate vascular beds [1-AR in mesenteric resistance arteries from CHF rats, it was confirmed that Ca2+ plays a more crucial role in the vascular contractility as CHF develops [2+ is not due to changes in Ca2+ sensitivity, nor in intracellular Ca2+ release, but rather to changes in the α1-AR-triggered Ca2+ influx [Disease progression might also modify the dependency of αstage HF ,120. Mordevelops . Further+ influx .1-AR vasoconstriction to Ca2+. For instance, in CHF rats an increased responsiveness of CHF femoral resistance arteries was identified which appeared to be the result of first, a RhoA/Rho kinase-depended increase in sensitivity to Ca2+ and second, of the IP3-induced increase in intracellular Ca2+ [2+ was reported to be decreased in femoral resistance and muscle resistance arteries of CHF rats, which presented as a dissociation between the intracellular Ca2+ and the generated wall tension [Other studies, however, have identified a modification in the sensitivity of αlar Ca2+ . On the  tension .1-ARs constitute an important homeostatic mechanism that contributes to the maintenance of peripheral vascular resistance at normal conditions and also in response to shock. Nevertheless, α1-AR regulation and function might alter due to disease. Even though HF is characterized by enhanced vascular resistance [1-AR stimulation in HF vessels.αsistance , there i1-AR and consequently, in the failure of the contractile apparatus to function properly. Attenuation of α1-AR responsiveness after continuous exposure to catecholamines has been validated in various animal studies [1-AR mediated vascular responses in both animals and humans with HF, the findings are not unanimous . Moreover, a cardioselective approach in the case of α1 agonism appears to be crucial for the exploitation of only its advantageous characteristics by avoiding potential vascular side effects. A subtype approach could also be beneficial but further studies are necessary in order to gain more insight in such treatment techniques. Finally, in view of the positive outcomes from clinical studies with carvedilol, more extensive investigation is to be expected so as to better comprehend the pharmacologic properties and mechanism of action of this agent. It is, therefore, essential that we expand our knowledge, and we apprehend current inconsistencies in order to pave the way to safer and more effective therapies.In conclusion, more research is required to further understand the complex mechanisms of α


Despite the improvement in the treatment options for prostate cancer patients, we still lack effective treatments when the cancer has spread outside the prostate. Therefore, there is an urgent need to develop new therapies that will work better in this setting. Our study has identified one central factor, the serum response factor (SRF), which controls many genes associated with prostate cancer development. We have shown that an SRF inhibitor can stop the cancer cells from dividing and surviving. When this inhibitor is used in combination with current treatments, these are more effective in killing cancer cells. We also confirmed the relevance of SRF to patients by looking at its abundance in prostate cancer tissues from patients. We showed that patients who did not respond to current treatments had significantly higher levels of SRF. We are currently investigating SRF inhibitor mechanisms and hope that these drugs will soon be available to patients.Castrate-resistant prostate cancer (CRPC) is challenging to treat with the androgen receptor (AR), the main target and key focus of resistance. Understanding the mechanisms of AR interaction with co-regulators will identify new therapeutic targets to overcome AR resistance mechanisms. We previously identified the serum response factor (SRF) as a lead target in an in vitro model of CRPC and showed that SRF expression in tissues of CRPC patients was associated with shorter survival. Here, we tested SRF inhibition in vitro and in vivo to assess SRF as a potential target in CRPC. Inhibition of SRF with the small-molecule inhibitor CCG1423 resulted in enhanced response to enzalutamide in vitro and reduced tumour volume of LuCaP 35CR, a CRPC patient-derived xenograft model. Nuclear localisation of AR post-CCG1423 was significantly decreased and was associated with decreased α-tubulin acetylation in vitro and decreased prostate specific antigen (PSA) levels in vivo. SRF immunoreactivity was tested in metastatic tissues from CRPC patients to investigate its role in enzalutamide response. Kaplan–Meier curves showed that high SRF expression was associated with shorter response to enzalutamide. Our study supports the use of SRF inhibitors to improve response to enzalutamide. Current treatments for locally advanced and metastatic prostate cancer are mainly focused on targeting the androgen receptor (AR). However, despite initial response, most men progress to develop castrate-resistant prostate cancer (CRPC), which is challenging to treat. Recently, second generation AR blockers, such as abiraterone acetate and enzalutamide, have been introduced into the clinic, but resistance to these treatments also develops ,2,3,4. TOur group and others identified the serum response factor (SRF), a transcription factor involved in cytoskeleton organisation and cellular proliferation , as an iThe relevance of SRF protein expression in prostate cancer patients was demonstrated in several published studies ,12,13,14Due to the potential use of SRF as a therapeutic target in advanced prostate cancer and its relationship with AR, in this study, we tested the combination treatment of enzalutamide and CCG1423 in vitro and in LuCaP 35CR, a patient-derived xenograft (PDX) model of enzalutamide resistance. Similarly to one recognised enzalutamide mechanisms of action , inhibitLNCaP parental and Abl isogenic pair cells were treated with six increasing concentrations of enzalutamide and CCG1423, singly and in combination for 5 days, and cell viability was assessed using 3--dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assays. This analysis showed that both cell lines are sensitive to CCG1423 with similar half-maximal inhibitory concentration (IC50) values . However, while LNCaP parental cells show lower IC50 values in response to enzalutamide compared to CCG1423 , the Abl cells, a model for castrate resistance, were more responsive to CCG1423 than enzalutamide with IC50 values of 9.97 ± 2.19 vs. 24.27 ± 2.28, respectively. Moreover, in both cell lines, the addition of CCG1423 to enzalutamide treatment resulted in a significant decrease in enzalutamide IC50 values with increasing concentrations of CCG1423 .p < 0.05), CCG1423 (p = 0.057) and combination (p < 0.05) treatments compared with controls (p < 0.01), CCG1423 (p < 0.01) and combination (p < 0.01) groups compared with controls  0.16)  were observed in the treatment groups  in comparison with the control group A. TV at controls B. In lincontrols C. While R) 0.16) D, while R) 0.16) .p < 0.01 for CCG1423 and p < 0.05 for the combination) (p < 0.05 for both CCG1423 single and combination treatment). The effect of CCG1423 on AR sub-cellular localisation was also confirmed in vitro in the LNCaP Abl cells (cellular model of CRPC). Immunocytochemistry (ICC) experiments showed decreased AR nuclear localisation following CCG1423 treatment, both as a single agent and in combination with enzalutamide  . This anination) B, while ination) C. Importlutamide E–H. Thesp = 0.039 for parental and 0.0003 for Abl). In the presence of DHT, a significant decrease in acetylation of α-tubulin was shown for LNCaP parental (p = 0.05) (p = 0.077)  A, while = 0.077) B. To con= 0.077) C. Since One mechanism of action of enzalutamide is to inhibit AR translocation to the nucleus by binding AR ligand binding domain, competing with DHT and inhibiting AR dimerisation, necessary for nuclear translocation . ConsideDespite the emergence of next-generation anti androgens such as enzalutamide and abiraterone acetate, advanced prostate cancer is still challenging to treat due to development of CRPC and subsequent resistance to other available treatments, such as docetaxel. AR mutations and splice variants are among the main mechanisms of resistance to enzalutamide . In an eThe small molecule inhibitor CCG1423  was usedSince PSA decrease is linked to a disruption in AR transcriptional activity, and a relationship between AR and SRF has been previously shown ,10,11, wSRF expression in tumour tissues from prostate cancer patients has been extensively characterised by our group, showing that elevated nuclear SRF expression occurs early on during prostate cancer progression  and is a2 in air.The LNCaP parental cells (ATCC) were cultured in Advanced RPMI-1640 medium supplemented with 10% Fetal Bovine Serum (FBS), 100 µL/mL streptomycin, 100 U/mL penicillin and 1% Hepes. The LNCaP Abl subline is a model for castrate resistance, with a 30-fold increase in AR transcriptional activity compared with the LNCaP parental line. LNCaP Abl cells were generated from the parental cell line as described previously  and cultThree thousand cells per well were seeded in 96-well plates. The following day the cells were treated either with vehicle (DMSO), increasing concentrations of CCG1423 (MedChem Express) alone , increasing concentrations of enzalutamide (MedChem Express) alone  or all the 36 possible combinations of CCG1423 and enzalutamide. Cells were treated for five days and cellular viability was measured by MTT assays as previously described . IC50 va3, animals were randomised to receive either vehicle control (n = 11), enzalutamide , CCG1423  or enzalutamide plus CCG1423 (dosing regimen as previously noted). Blood was collected from the retro-orbital sinus 3 weeks after the beginning of the treatment to determine serum PSA levels. Animals were sacrificed at 6 weeks post enrolment, when tumours exceeded 1000 mm3, or when they became otherwise compromised. Tumour volumes (TV) and body weights (BW) were measured twice weekly. Average PSA values and TV for each treatment group were compared using t test assuming equal variance. * p < 0.05, ** p < 0.01. Kaplan–Meier cumulative survival curves were generated using Graphpad, Prism program. Thirteen animals whose death was not related to tumour growth (found dead or had ulcerating tumours) were excluded from the survival analysis. Five-fold increase in TV from the enrolment was considered as “death”. Hazard ratios from Mantel–Haenszel models and p-values from the relevant log rank tests were calculated.The in vivo PDX experiment was approved by the University of Washington Institutional Animal Care and Use Committee (IRB number is IRB# 2341). Fox Chase CB17 SCID castrated male mice were implanted with LuCaP 35CR, a model of enzalutamide-resistant CRPC . Once tuFor IHC staining, antigen retrieval of the deparaffinised tissue sections was performed using a PT Link module (DAKO) at 95–99 °C for 20 min in a citric acid buffer ; this step was not necessary for ICC staining. Staining was performed using a DAKO autostainer Link 48 according to the manufacturer instructions. SRF antibody  and AR antibody (Santa Cruz AR (441): sc-7305) were used at a concentration of 1:500.p < 0.01, * p < 0.05.Following IHC, slides were scanned using an Aperio ScanScope XT system (Aperio Technologies) at 20× magnification. AR immunoreactivity was analysed by automatic quantification using Visopharm’s Oncotopix ; the classification method used “Cell Classification” for nuclear and cytoplasmic assessment. AR expression was reported as H-Score . AverageWestern blots were carried out as previously published . The folHuman tissue microarrays were constructed consisting of soft tissue metastases and bone metastases from 45 patients with advanced prostate cancer. Samples were obtained from patients who died of metastatic CRPC and who signed written informed consent for a rapid autopsy, under the aegis of the Prostate Cancer Donor Program at the University of Washington . Two repp-values from the relevant log rank tests are also displayed.Cores with necrosis were excluded from our analysis. Nuclear immunoreactivity for SRF in soft tissue metastases and bone metastases was assessed by automatic quantification, using Visopharm’s Oncotopix ; the classification method used “Threshold”. SRF expression was reported as H-Score. For the purpose of statistical analysis, the nuclear scores of SRF was divided into low and high expression. The cut-off values for low/high expression were calculated by dividing the patient H-Score data into quantiles, 1–99 in turn, and establishing the largest separation using the log-rank test. Kaplan–Meier cumulative survival curves were generated by dividing the data in two at the appropriate cut-off point, and assessing the times to the relevant endpoints, using the survival package (version 2.41–3)  in R (veOur study showed an alternative way to interfere with the AR signalling pathway, through inhibition of SRF. Considering that SRF expression has been associated with prostate cancer progression and resistance to enzalutamide in patients, we propose to target SRF as a therapeutic strategy to treat CRPC.


The first intelligent COVID-19 lockdown resulted in radical changes within the tertiary educational system within the Netherlands. These changes posed new challenges for university students and many social welfare agencies have warned that it could have adverse effects on the social wellbeing (SWB) of university students. Students may lack the necessary social study-related resources (peer- and lecturer support) (SSR) necessary to aid them in coping with the new demands that the lockdown may bring. As such, the present study aimed to investigate the trajectory patterns, rate of change and longitudinal associations between SSR and SWB of 175 Dutch students before and during the COVID-19 lockdown. A piecewise latent growth modelling approach was employed to sample students’ experiences over three months. Participants to complete a battery of psychometric assessments for five weeks before the COVID-19 lockdown was implemented, followed by two directly after and a month follow-up. The results were paradoxical and contradicting to initial expectations. Where SSR showed a linear rate of decline before- and significant growth trajectory during the lockdown, SWB remained moderate and stable. Further, initial levels and growth trajectories between SSR and SWB were only associated before the lockdown. The diverse biological, genetic, and epidemiological attributes of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or ‘COVID-19’) has made it one of the most contagious diseases in modern history  and mental health of individuals; particularly for vulnerable groups  increase dramatically during times of uncertainty  assessment occurred one month after the seventh assessment. The data was collated, captured, and stored on a secured server in compliance with the research institution’s data management policy.Attention Checks. Two of Abbey and Meloy’s (2017) guidelines for attention checks were implemented. Firstly, direct queries were inserted into the instructions of two sections of the questionnaire . Secondly, a post hoc analysis of the response patterns, response consistency and time taken to complete the questionnaire was implemented  and required participants to complete a battery of psychometric assessments for five weeks c.f. Table Most of the participants were Dutch-speaking (94.3%), Dutch national (94.3%) males (66.3%) between the ages of 22 and 25 years old (94.3%) enrolled for a master’s program at a University in the Netherlands  and lecturer-support (8 items: e.g. ‘Can you discuss study problems with your lecturers?’). Participants were requested to reflect upon the preceding week and rate items on a 5-point Likert scale ranging from 1 (“Never”) to 5 (“Always”). The scale showed to be a reliable instrument across all 8-time points in this study with Cronbach’s ranging from 0.84 to 0.96.The Social Wellbeing Subscale of the Mental Health Continuum Short-Form . The scale showed to be a reliable instrument across all eight-time points in this study with Cronbach’s ranging from 0.76 to 0.85.The rm Keyes  was usedFirst, the distribution of the data, the level of internal consistency and the relationships amongst the factors were assessed through descriptive statistics  and Pearson/Spearman correlations. Skewness and Kurtosis ranging between − 2 and + 2 were used as indicators of multivariate normality  and effect sizes were either small (r > 0.10), medium (r > 0.30) or large (r > 0.50)  and Mplus v.8.4  Steyn , 2002.Second, through structural equation modelling (SEM) with the robust maximum likelihood (MLR) estimator, a series of unconditional Latent Growth Models (LGM) were estimated to determine the intercept and slopes of students’ SSR and SWB. A sequential and competing measurement model process was employed to determine the best fitting LGM. First, an intercept only model for each factor was estimated. Thereafter separate LGM for linear-, quadratic-, and piecewise growth trajectories were calculated to determine the best-fitting model for the data. For the piecewise LGM, two separate linear growth factors were estimated representing the slopes before (Week 0–4) and during (Week 5–10) the COVID-19 lockdown. Time 4 represented the interior knot  with effect sizes ranging from small (r = 0.14) to large (r = 0.83).Table p > 0.05; SRMR < 0.08) and we could therefore proceed to estimate and compare the competing LGMs.Before estimating and comparing different latent growth models, a series of confirmatory factor analytical models for SSR and SWB, at each measurement instance, were estimated. Here, each item was estimated to load onto its a priori theoretical factorial model. Observed items were used as indicators for latent factors and no items were omitted. With the exclusion of Week 2 and 4 on the SWB scale, no error variances on items were permitted to correlate. Items 2 and 4 for both Week 2 and 4 on the SWB scale were allowed to co-vary to improve overall model fit. The results, summarized in Table 2N=175) and SWB (χ2N=175). Two growth trajectories were confirmed for both factors: Phase 1: before the lockdown procedures (Time 0–4) and Phase 2: during lockdown (Time 5–7). The knot was set at Time 4.A series of LGM were estimated to find the best fitting model for both SSR and SWB. Comparing an intercept-only, linear-, quadratic-, and piecewise unconditional LGM presented in Table ssr = 3.579, S.E. = 0.04, p < 0.05) and SWB . This implies that at the start of the semester students experienced average levels of SSR and SWB. Further, both intercepts’ variances were significant, showing inter-individual variability. Individual growth trajectories significantly differed from one another around the estimated mean for both SSR and SWB.Further, the unstandardized estimates in Table 1ssr = -0.016, p < 0.05) but during lockdown increased by 0.01 base points week-on-week . However, both the latent growth trajectories for SWB were non-significant  implying that it remained relatively constant throughout the study period. Figures Both growth trajectories (i.e. the Slopes) for SSR were significant. This implies that before the COVID-19 lockdown, SSR decreased linearly  = -0.012, p < 0.05) and SWB  = -0.029, p < 0.05) was significantly negatively related to their respective starting values. In other words, those who reported high at baseline had a slightly faster rate of decline before the lockdown measures.The significant co-variances between the Intercept and Slope 1 of SSR as well as that of SWB indicates that the rate of change in SSR (p > 0.05) implying that the rate of change during lockdown was not dependent upon their initial values.All the remaining covariances between slopes and intercepts for both the SSR- and SWB models were non-significant (2N=175). The intercept of SSR (Issr) was positively associated with baseline levels of SWB (Iswb)  which implies initial levels of the former directly influenced initial levels of the latter. Further, the initial growth trajectory of SSR (S1ssr) before the COVID-19 lockdown procedures, predicted the initial rate of change in SWB (S1swb) .Figure Cov =  −.36, S.E.:0.10). No other associations between the intercepts and slopes for either SSR or SWB was found. The results imply that both SSR and SWB changed at different rates and in different directions after lockdown. Where SSR decreased before-  but increased after lockdown , SWB stayed moderate and stable before and during lockdown .Finally, only the covariance between the Intercept and pre-lockdown Slope of SSR were significant showing that higher starting levels were associated with a faster decline before lockdown . Access to lecturers via electronic  increased significantly, and this may have helped to contain the anxiety associated with the changes in educational activities. Secondly, with the new online educational models being introduced, students may have experienced more autonomy in attending to educational activities (e.g. viewing video lectures when convenient to them), and it may have been easier to negotiate assignment deadlines/exam content with lecturers. This may have strengthened perceptions of lecturer support as educational activities are usually highly structured with limited scope for major changes during the progression of a course  . Therefore, the experiences reported may have differed from those of other universities in the country. These results may therefore not be generalizable. Thirdly, the measurement instances during lockdown only spanned a few weeks, and students may still have been coming to terms with the “new normal”. Therefore, additional measures during lockdown may have painted a more holistic picture. Fourthly, the study focused on group related changed and did not explore specific inter-individual differences. This paper is the first to look at how trajectories of social wellbeing and social student resources change before and during the COVID-19 outbreak in the Netherlands. It provides valuable information which could be used to understand how pandemics actively affect components of mental health. The findings present an alternative view of the current narrative within the literature as to the negative impact of the COVID-19 pandemic on the social wellbeing of students.


MDHAR function and regulatory mechanism in Gossypium have been limited. Herein, a genome-wide identification and comprehensive bioinformatic analysis of 36 MDHAR family genes in four Gossypium species, Gossypium arboreum, G. raimondii, G. hirsutum, and G. barbadense, were performed, indicating their close evolutionary relationship. Expression analysis of GhMDHARs in different cotton tissues and under abiotic stress and phytohormone treatment revealed diverse expression features. Fiber-specific expression analysis showed that GhMDHAR1A/D, 3A/D and 4A/D were preferentially expressed in fiber fast elongating stages to reach peak values in 15-DPA fibers, with corresponding coincident observances of MDHAR enzyme activity, AsA content and ascorbic acid/dehydroascorbic acid (AsA/DHA) ratio. Meanwhile, there was a close positive correlation between the increase of AsA content and AsA/DHA ratio catalyzed by MDHAR and fiber elongation development in different fiber-length cotton cultivars, suggesting the potential important function of MDHAR for fiber growth. Following H2O2 stimulation, GhMDHAR demonstrated immediate responses at the levels of mRNA, enzyme, the product of AsA and corresponding AsA/DHA value, and antioxidative activity. These results for the first time provide a comprehensive systemic analysis of the MDHAR gene family in plants and the four cotton species and demonstrate the contribution of MDHAR to fiber elongation development by controlling AsA-recycling-mediated cellular redox homeostasis.Monodehydroasorbate reductase (MDHAR) (EC1.6.5.4), a key enzyme in ascorbate-glutathione recycling, plays important roles in cell growth, plant development and physiological response to environmental stress via control of ascorbic acid (AsA)-mediated reduction/oxidation (redox) regulation. Until now, information regarding  As a byto water ,9.myo-inositol (MI)-dependent d-glucuronate pathway, the d-galacturonic acid pathway and the recycling pathway by the Foyer–Halliwell–Asada cycle [LeMDHAR gene significantly elevated AsA level and decreased the H2O2 content in tomato (Lycopersicon esculentum Mill.) [Chlamydomonas reinhardtii CrMDHAR1 is a ROS-inducing gene and its overexpression elevated the AsA level and AsA/DHA ratio to resist photooxidative stress [AtMDHAR1 correlated enrichment of MDHAR enzyme activity and accumulation of AsA content with significantly enhanced tolerance against environmental stress [As evinced by genetic experiments, Asc is an important abundant antioxidant that executes a vital role in detoxifying cellular excessive ROS by enzymatic reactions ,9, regulda cycle ,18,19. Ada cycle ,21. Atteda cycle . Overexpm Mill.) . Chlamyde stress . Transgel stress .MDHAR cDNA has been cloned from diverse plant species containing pea (Pisum sativum L.) [Lycopersicon esculentum Mill.) [Spinacea oleracea) [Cucumis sativus), soybean (Glycine max) and potato (Solanum tuberosum) [Spinacia oleracea) [Physcomitrella patens PpMDHAR cytosolic isoforms indicated different response patterns with increased accumulation of PpMDHAR1 and PpMDHAR3 to salt and osmotic stresses, and normal level of PpMDHAR2 to chilling and oxidative stresses, showing that cytosolic MDHARs are essential for land plants to adapt to dry terrestrial conditions [As a thiol-group-containing flavin adenine dinucleotide (FAD) monomeric enzyme of the AsA-GSH cycle, MDHAR belongs to a multigene family and has been reported to locate in different organelles such as the chloroplast ,26, mitoivum L.) , tomato m Mill.)  and spinleracea) . The MDHberosum) ,36,37, tberosum) , and chlleracea) . Three Pnditions . In Arabnditions .Saccharomyces cerevisiae fermentative capacity [Numbers of reports have revealed the important role of MDHAR in resisting abiotic stress in different plants such as arabidopsis , wheat , tomato capacity , strengtcapacity , as wellcapacity , postgercapacity  and fruicapacity .GhVTC1 is the key factor to promote cell elongation under control of ethylene [myo-inositol-1-phosphate synthase (MIPS) located in the MI-dependent d-glucuronate pathway participated in cell elongation by providing the precursor MI to contribute to Asc biosynthesis [2O2-mediated apoplast oxidation [MDHAR genes and their functions, as well as the regulatory mechanism involved in fiber development in Gossypium, are largely unknown. In the present study, by a genome-wide identification in four cotton species of G. arboreum, G. raimondii, G. hirsutum and G. barbadense, a total of 36 MDHAR genes were obtained. A comprehensive systematic bioinformatic analysis, including chromosomal location, gene structure, motif distribution and evolutionary relationships, was performed indicating rapid evolution and close evolutionary relationships. G. hirsutum GhMDHARs that showed diverse expression patterns under abiotic stress, and phytohormone treatment demonstrated a close correlation with fiber growth by increasing MDHAR activity, AsA content and AsA/DHA ratio, possibly via involvement of AsA-mediated H2O2 signaling pathway.Cotton cultivars are important crop plants worldwide for their supply of fibers used as raw materials to the textile industry. Fiber growth and development is closely correlated with the final fiber quality ,48. A geethylene , while tynthesis . Ascorbaxidation . TogetheG. arboreum, G. raimondii, G. hirsutum and G. barbadense using arabidopsis MDHAR proteins as queries with the BlastP procedure, a total of 36 MDHAR genes were identified, with six in diploid cotton (G. arboreum and G. raimondii), and twelve in allotetraploid cotton (G. barbadense and G. hirsutum) (MDHAR members in comparison to the diploid cotton species. The MDHARs possess open reading frame (ORF) lengths ranging from 1305 to 1677 base pair (bp), the encoded proteins have 434 to 558 amino acids with theoretical molecular weight (MW) from 46.83 to 62.25 kDa and theoretical pI from 5.9 to 9.06, respectively (By searching the genome of irsutum) . The allectively .MDHARs were distributed across different chromosomes, with locations on fivechromosomes of G. arboreum  and of G. raimondii . The distributions in allotetraploid cotton species showed similar patterns of orthologous MDHARs on the same chromosomes of G. barbadense and G. hirsutum, with locations on five At and five Dt subgenomes  , were found to be segmentally duplicated. The above four paralogous gene pairs were also identified as duplications in the Dt subgenome, with the absence of MDHAR3-MDHAR6 in the G. barbadense Dt subgenome (The  and 13) . The genubgenome .MDHAR sequences from four cotton species of G. arboreum, G. raimondii, G. barbadense and G. hirsutum, and from another eight plants including Glycine max, Oryza sativa, Sorghum bicolor, Zea mays, Arabidopsis thaliana, Theobroma cacao, Vitis vinifera and Cicer arietinum, were used for construction of a neighbor-joining (N—J) tree, resulting in three typical groups (Group A–C). The orthologous MDHARs in cotton and cacao indicated a close relationship, which might be the reason that they are in the same family of Malvaceae. Cotton MDHAR homologs were classified into the same branches, with MDHAR1, 3, and 6 in group A, MDHAR 2 and 5 in group B, and MDHAR 4 in group C  indicated that the ratio of all duplicated MDHAR gene pairs showed a value of <1, indicating all these MDHAR gene pairs were purifying selection and suggesting their rapid evolution  and Multiple Em for Motif Elicitation) (MEME) programs respectively. The MDHAR genes could be divided into different branches . The results indicated that, during different fiber developmental stages, GhMDHAR3A/D showed the most significant accumulations in 5–21 days post anthesis (DPA) fibers (the fast elongation stages), with the peak value at 15 DPA. GhMDHAR1D also presented moderate upregulated expression levels in 5–21 DPA fibers. GhMDHAR1A and GhMDHAR4A/D were mainly enriched in 15-DPA fibers  and NaCl and plant hormones of IAA and ETH treatments were used, and expression abundance of GhMDHAR genes was detected by qRT-PCR, using 0-h expression level of untreated samples as control. The GhMDHAR genes exhibited diverse expression patterns under different treatment conditions. GhMDHAR2A/D, GhMDHAR5A/D, and GhMDHAR6A/D indicated significant accumulated expression under these four treatments, at least in one treated time point, with the exceptions of GhMDHAR5A/D that showed moderate enrichment and of GhMDHAR6A/D that also showed moderate regulation under IAA treatment  indicated significant promotions after 12-h H2O2 stimulation  [G. barbadense  [G. hirsutum  [G. raimondii  [G. hirsutum were downloaded from NCBI (https://www.ncbi.nlm.nih.gov) SRA database (PRJNA248163). The RNA-Seq clean data were mapped to the presented Gossypium genome by TopHat2  and the fragments per kilobase of exon per million reads mapped (FPKM) value was calculated by Cufflinks  [http://www.ebi.ac.uk [pI) were calculated by online tools at ExPASy website (https://www.expasy.org). Subcellular localizations of the MDHAR proteins were predicted by the online tool ProtComp at Softberry server (http://www.softberry.com/). The MDHAR chromosomal distributions were described by Mapinspect software .The cotton MDHAR sequences were obtained by submitting arabidopsis MDHARs to the genome database for genome/) , G. barben.org/) , G. hirsportal/)  and G. rportal/)  using loia, USA) ,86. The bi.ac.uk . The thehttp://gsds.cbi.pku.edu.cn/index.php) and Multiple EM for Motif Elicitation  respectively [Ka/Ks was calculated by Dnasp 5.0 software  [Ka/Ks ratios for the GhMDHAR genes were calculated to assess the selection pressure on duplicated genes, with Ka/Ks ratio >1, <1, or = 1 to indicate positive selection, negative selection and purify evolution, respectively [http://bioinformatics.psb.ugent.be/webtools/plantcare/) for analyzing the cis-acting element composition [Multiple sequence alignment of the MDHAR proteins was performed using ClustalW and the phylogenetic tree of the MDHAR proteins from four cotton cultivars and another eight plant species was constructed by Molecular Evolutionary Genetics Analysis  with the parameters of neighbor-joining (N—J) method of 1000 bootstrap replicates, model of p-distance, and Gaps/Missing data treatment by Pairwise deletion. The exon-intron distribution and conserved motif analyses were recognized using the online software of the gene structure display server  . The Ka/ectively . The Tajectively . The TBtposition .G. hirsutum) varieties  and Sea-island cotton (XH36) plants were cultivated in the experimental field of Shihezi University in Shihezi city, Xinjiang, China. The materials of X142 were collected for gene expression analysis, enzyme activity measurement, and treatments of stress and phytohormone. Cotton bolls of X142 were marked with a label on the day post anthesis (defined as 0 DPA) and the ovules and fibers were then collected in different developmental time points of 5, 10, 15, 18, 21 and 28 DPA for cryopreservation and successive use. Cotton seeds were sown in nutrient soil and planted in the greenhouse at 28 °C with a photoperiod of 16 h light/8 h dark. The leaves of three-week-old seedlings of X142 were treated with 20% polyethylene glycol (PEG), 100 mM sodium chloride (NaCl), 100 µm indole-3-acetic acid (IAA), 100 µm ethephon (ETH), and 100 mM H2O2 for 0, 6, 12, 24 h respectively. The treated leaf materials of X142 were collected and stored at −80 °C after flash-freezing in liquid nitrogen for further utilization. The matured fibers of different cotton varieties  were collected for length measurement by three independent experiments with 30 samples for each replicate.Upland cotton , and then used as a template for qRT-PCR detections with SYBR green real-time PCR master mixes . The specific primers were provided in −ΔΔCt method using the GhUBQ7 gene as internal control for normalization [GhMDHAR genes measured by qRT-PCR were used to generate a heatmap that visualized by MultiExperiment viewer  software.Total RNA was extracted from different cotton tissues by our previous method as described ,16. The lization ,95. The 2) that was then centrifuged at 14,000 rpm at 4 °C for 10 min. The reaction mix was obtained by the addition of collected enzyme extracts in pH 7.6 50 mM potassium phosphate buffer , and enough ascorbate oxidase (0.5 unit) was added into the mix solution to trigger the reaction. The values were detected at 340 nm for 2 min, and the absorbance decrease generated by the oxidation of NADH was used to calculate the MDHAR enzyme activity with the extinction coefficient of 6.22 mM−1·cm−1 [The MDHAR enzyme activity was measured by a spectrophotometric method. The materials were ground to powder in liquid nitrogen, and about 50 mg of raw powder was added into 1 mL of pH 6.0 50 mM potassium phosphate buffer  was added to the reaction mix for total Asc determination.Asc measurement was performed as in our previous method . Briefly


Idiopathic facial paralysis, also known as Bell's palsy, exerts a negative effect on the quality of life. Although the prognosis is good in the majority of cases, a significant percentage of affected individuals may have sequelae that can negatively affect their lives. The use of therapeutic measures as early as possible can improve the prognosis. This article describes the successful use of laser-photobiomodulation as a single therapy in a patient with Bell's palsy and confirms the possibility of using this therapeutic modality as a good choice, since it is a therapy that is painless, comfortable, and without systemic side effects. The findings demonstrate that the adequate use of laser-photobiomodulation can be an effective therapeutic option for patients with Bell's palsy, regardless of the age, shortening the recovery time obtained with conventional therapies and avoiding sequelae. Further studies are needed for the establishment of adequate protocols. Idiopathic facial paralysis, also known as Bell's palsy, is the most common form of facial paralysis, accounting for 60 to 75% of cases . This coThe etiology of this condition remains unclear –5. HowevThe diagnosis can be obtained through clinical examination, but complementary exams can be useful, especially for the elucidation of the differential diagnosis in relation to certain diseases, such as Ramsay Hunt syndrome , 8, LymeThe incidence of Bell's palsy is between 23 and 35 cases for every 100,000 people –13. The Different treatments have been proposed to achieve rapid recovery without significant sequelae. Such treatments include botulinum toxin , acupuncThe occurrence of Bell's palsy during pregnancy  requiresSome studies have been conducted to predict the prognosis and quantify the degree of impairment in Bell's palsy , 20. Altth, 2018. After cranial tomography for the differential diagnosis in relation to a possible transitory stroke, the neurologist diagnostic was Bell's palsy. In our first evaluation . She sought an emergency service on March 4 Figures . This cl™) was previously measured to have a 100 mW output power density with a fiber diameter of 600 μm. The protocol was punctual application of 3.3 J per point (120 J/cm2), 10 seconds each, in contact with the skin, infrared of 808 nm, and applied to ten points .Bell's palsy is generally a self-limiting condition, with the return of functions within a six-month period and with no substantial sequelae in most cases. However, a significant portion of patients have more severe conditions. It is estimated that 71% of affected individuals recover completely, about 13% only achieve partial recovery, and approximately 4% experience severe paralysis [The conventional therapies include drug therapy , facial exercises, massage, thermotherapy, electrical stimulation, acupuncture, and laser-photobiomodulation –4, 15.Regarding the recovery time, the literature shows that, with conventional therapies, 69.5% of patients have good recovery after 3 months, while 30.5% have poor results .Recovery results are often evaluated by the House-Brackmann Scale, assessing the levels of facial nerve injury and proposing the following order: normal, soft, moderate, moderate/severe, severe dysfunction, and complete paralysis , 21.Some studies have demonstrated that L-PBM enhances the regeneration of neurons, with local and systemic effects, directly restoring nerve structure and their communication with the central nervous system. Some researchers have demonstrated that laser increases microcirculation, activating angiogenesis and stimulating nerve regeneration . Laser-pA recent study reports that oxidative stress is high among individuals with Bell's palsy . As one Idiopathic facial paralysis  is an event that has a benign course in the majority of cases but can leave sequelae and have negative social, professional, and psychological impacts. Apparently, the early institution of therapeutic measures improves the chances of a complete recovery. Laser-photobiomodulation is a promising treatment option in such cases. However, controlled clinical trials are needed to enable the establishment of safe, adequate, and individualized protocols.


The most frequent cause of readmission was pyelonephritis (11.9%), followed by pneumonia (10.9%), compression fracture (10.6%), heat stroke (8.4%), and cerebral stroke (8.0%). In conclusion, older Japanese patients discharged from rehabilitation units have lower readmission rates than those previously reported. Thus, better nutritional control, a multidisciplinary approach to the management of cognitive dysfunction, and a decrease in polypharmacy could be associated with improved outcomes among discharged older patients.Rehabilitation for hospitalized older people can improve their independence for performing activities of daily living (ADL), but determining its appropriateness can be challenging because of inherent limitations in their ADL and short life expectancy. Thus, we aimed to clarify the benefit of rehabilitation among older Japanese patients. We retrospectively evaluated consecutive older patients (age > 65 years) admitted to the rehabilitation unit of a rural community hospital between 1 April 2016 and 31 March 2020. The primary outcome measure was readmission for acute conditions. Of the 732 patients evaluated, 311 patients (42.5%) were readmitted. Readmission was significantly associated with body mass index (BMI) ( Rehabilitation is an important aspect of the continuity of home care among admitted older people as it can improve independence in their performance of activities of daily living (ADL) in their homes . RehabilA challenge in the rehabilitation of older patients is determining the appropriateness of continuing rehabilitation because of the possibility of a limited benefit in improving ADL and the short life expectancy in this population. Readmission to acute care hospitals is an established outcome measure of rehabilitation ,13. HighA longer duration of rehabilitation is related to a higher readmission rate ,18. AccoThe outcome of rehabilitation after discharge for older people is critical for the decision to continue rehabilitation. Most patients undergoing rehabilitation in rural community hospitals are aged over 80 years and are frail with multiple morbidities ,22,23. RThis was a retrospective cohort study of consecutive patients aged over 65 years who were admitted between 1 April 2016 and 31 March 2020 to Unnan City Hospital for acute diseases and transferred to the rehabilitation unit. Unnan City is a rural city located southeast of the Shimane Prefecture in Japan. In 2020, the total population was 37,638 . Older individuals (age > 65 years) account for 39% of the population, and this is projected to increase to 50% by 2025 .The patients were regularly followed up at Unnan City Hospital or other medical institutions in Unnan City from April 2016 to 30 October 2020. All readmissions were at Unnan City Hospital. All participants were followed until readmission or death .The recovery rehabilitation unit of Unnan City Hospital had 30 rehabilitation beds during the study period. The unit accommodated patients motivated to return home after rehabilitation. Most of the patients had underlying internal medicine or orthopedic conditions. The rehabilitation plan was discussed with the patients and their family by a physician and the chief nurse in charge of the recovery rehabilitation unit. The decision to move from acute care to the recovery rehabilitation unit was undertaken collaboratively. Rehabilitation was performed at an average of twice per day (60–90 min per session) by physical and occupational therapists. If the patient had swallowing and speaking problems, speech therapists were involved. The discharge timing and location were decided based on discussions among patients; their families; and a team consisting of the attending physician, nurse, and social workers. The team is specialized in providing support and decision-making for patient discharge .Patient information was extracted from the electronic medical records of Unnan City Hospital throughout the research period. The main outcome measure was readmission to the hospital after discharge from the rehabilitation unit. Planned readmissons for chemotherapy and surgeries were excluded. The following data were collected: age; sex; body mass index (BMI); serum albumin (g/dL) as an indicator of nutritional status; reasons for readmission; the number of medications used (to assess polypharmacy) ; the Chap-value of <0.05 was considered statistically significant.Parametric data were compared using the Student’s t-test, while nonparametric data were analyzed using the Mann–Whitney U test. Based on previous studies and the average of variables, numerical variables were dichotomized as follows: CCI, ≥5 and <5 ; care leThe hospital was assured of no loss of anonymity and confidentiality regarding the patients′ information. The information related to this study was posted on the hospital website without the disclosure of any details concerning the patients. To address any questions regarding this study, contact information of the hospital representative was also listed on the website. The Unnan City Hospital Clinical Ethics Committee approved this study .Of the 951 patients admitted to the rehabilitation unit, 845 patients were aged >65 years. After excluding 113 patients with missing data, 732 participants were finally evaluated. The patient inclusion flowchart is shown in p < 0.001), number of medicines (p = 0.014), CCI (p = 0.004), and dependent condition (p < 0.001) were higher in the readmission group.The average patient age was 84 years (standard deviation = 8.06), and 32.7% of the participants were male. A total of 311 participants (42.5%) were readmitted to the hospital. BMI (p = 0.003). Regarding FIM, all scores were statistically higher in the non-readmission group. The length of rehabilitation was longer in the readmission group , dependent condition (p < 0.001), higher cognitive domain scores of FIM (p = 0.019), and polypharmacy (p = 0.026) were significantly associated with readmission.Kaplan–Meier curves show the estimated probability of non-readmission as a function of total FIM and of the cognitive and motor components of FIM . A Cox rDetermination of the benefits of rehabilitation in older patients can be challenging because of the inherent limitations in their ADL and their short life expectancy. In this study, less than 50% of the patients who underwent rehabilitation were readmitted to the hospital within 4 years. Lower BMI, dependent conditions, lower cognitive domain scores in FIM, and polypharmacy were related to readmission after discharge from the rehabilitation unit. The most frequent causes of readmission were infections, including pyelonephritis and pneumonia.The lower readmission rate for patients admitted to rural hospitals can be partly attributed to the Japanese rehabilitation system and health insurance. The 30- and 90-day readmission rates in this study were much lower than those in previous studies ,35,36,372, had lower readmission rates. Accordingly, this population should aim to have a moderated BMI that is ≥22 kg/m2. To achieve better outcomes in rehabilitation units, the healthcare staff should also include the patient’s BMI in the rehabilitation plan, ensuring that patients with BMIs lower than 22 kg/m2 are provided interventions to increase their BMI through training and better nutrition.We also found that a higher BMI was associated with a lower risk of readmission. In older patients, a higher BMI can indicate better nutrition and muscle conditions. The relationship between BMI and mortality indicates that a higher BMI can result in better longevity . PreviouCognitive function at discharge and dependent condition were also identified as influencing factors for readmission. Low cognitive function can mask critical symptoms, leading to disease exacerbation and admission. Good cognitive function is important in older patients because it will enable them to effectively convey their symptoms. Some dependent older people with low cognitive function are isolated in rural areas; thus, home care professionals struggle to manage older people who cannot accurately report their symptoms ,45,46. FPolypharmacy is implemented before discharge in order to reduce the risk of readmission. Polypharmacy, which is common among older patients, can affect the physiology of older patients  and causThis study has some limitations. First, it was performed in a single rehabilitation center in a rural community hospital, and this may have affected the external validity. Future studies should investigate the outcomes of older patients in different types of hospitals in other countries. The second limitation was the low follow-up rate. Some patients were discharged to other cities and could not be followed-up; this affected the reliability of our findings. Despite these limitations, our findings provide data that may also be reflective of other settings in Japan. The data may be used as a base for developing relevant guidelines on the rehabilitation of older patients, including those in other countries, and prevent readmission.A higher BMI, no dependent conditions, a higher cognitive domain score in FIM, and no polypharmacy could be associated with the risk of readmission among older patients admitted to rehabilitation units. Thus, better nutritional control, collaboration among healthcare professionals for the management of cognitive dysfunction, and efforts to reduce polypharmacy are important for improving the outcomes of these patients.


It is well-known that no local model—in theory—can simulate the outcome statistics of a Bell-type experiment as long as the detection efficiency is higher than a threshold value. For the Clauser–Horne–Shimony–Holt (CHSH) Bell inequality this theoretical threshold value is  That is, if a Bell-type experiment is performed and the results show a violation of a Bell inequality, then the underlying physical process cannot be explained by a local theory. This kind of tests are called Bell tests and the violation of the inequality is called Bell violation. Since the earlier demonstrations utilizing cascade decays in atoms5, Bell violations have been observed in tests utilizing nonlinear optical processes9, ions10, neutral atoms11, Josephson junction12 and solid state qubits13. The implications of the Bell test not only change our understanding of nature, but also find application in device independent (DI) quantum communications16, randomness generation and amplification19, DI-verified quantum computation21, certifying quantum devices23 and DI bit commitment24. Entanglement, a necessary precondition for unconditional security26 in quantum key distribution, can also be certified from the violation of a Bell inequality, independently of the underlying implementation details. This paves the way for the device-independent tests of security28. However, for the observed Bell violation to be conclusive, it is important that the Bell test is loophole-free.More than 50 years ago, John Stewart Bell showed that any physical theory based on the assumptions of locality  and realism  must satisfy a set of statistical criteria called Bell inequalities29. In general, the detection loophole is a scenario in which the observed Bell violation (a test statistics) is no longer reliable as the measurement sample and may not be a true representative of the population . Crucially, this situation commonly happens in practice as practical detectors have finite detection efficiencies and hence one could end up with samples that are non-representative. While the detection loophole is not an issue for non-adversarial settings, the same is not true for the case of quantum cryptography since an adversary can take advantage of it to come up with a local model to fake Bell violations30. For this reason, much effort has been devoted to closing the detection loophole in practice.More specifically, a loophole-free Bell test is an entanglement experiment that requires multiple implementation loopholes such as the detection, locality, and measurement-independent loopholes to be closed simultaneously. Here, we focus on the detection loophole, and defer the rest to Ref.32. However, methods of experimentally implementing such correlations using practical means have been rarely discussed, despite its importance in practical quantum cryptography. The state-of-the art method is arguably that of Ref.30, where the authors demonstrated how an adversary could implement a local model using existing optical detector control methods to violate a Bell inequality for active basis choice schemes. However, their local model is effective only for a detector efficiency of up to 35 can be exploited to both fake the violation of the standard Clauser–Horne–Shimony–Holt (CHSH) Bell test all the way up to its threshold efficiency and simulate the local bound of the more general Garg–Mermin Bell test. Our results point out once again that when Bell tests are performed for certifying randomness, guaranteeing security in quantum communications, or detecting non-locality, they should either be performed with an efficiency at which the test is robust against detection loopholes, or should use the bound given by more general inequalities  when additionally exposed to a light pulse of intensity I equal to or higher than a threshold level I. For example, if the measurement basis matches that of the incoming light pulse, then all of it is incident on a single detector with intensity 47 and superconducting nanowires49. Next, we show how an adversary can exploit it to implement a local-realistic model.Conditions for violation: Let us assume that an arbitrary value of E requires the ratio of similar to different outcomes to follow Eq. , and no-detection outcome with probability At Alice, whenever the measurement basis matches (does not match) that of the incoming light, a deterministic “For details on how the above assumptions can be met in practice, please see the Methods. For each setting, the possible outcomes at Alice and Bob and the corresponding coincidence probabilities are shown in Table E is related to a and b asE corresponding to that a, b. As long as Eq. . For simplicity, let us assume the case when the adversary sends a light polarized at angle b/2, she sends the light at an angle midway between b/2. The condition for this is a; while when the basis mismatches, it results in a random outcome with probability b. This supports the practicality of our assumption. Note that this method leads to asymmetric detection efficiency, as Bob’s efficiency is always higher than Alice’s. However, this can be avoided by reversing the roles of Alice and Bob half of the time.47. Real detectors have noise, which leads to them having two thresholds 44. Generally, if the ratio I may allow an alternative method of controlling a and b, as we show below.We have so far assumed that the blinded detector is controllable as a step function: for I47. This can be used to obtain probabilistic detections. To illustrate this, we have measured control characteristics of one avalanche photodiode detector in a commercial QKD system Clavis254. At a particular continuous-wave blinding power, we varied the trigger pulse energy and recorded the corresponding click probability as shown in Fig. I (without varying the polarization by Practical detectors, when blinded, gradually increase their click probability from 0 to 1 in a certain range of trigger intensity


Prostate cancer is one of the most frequent cancers in men and is a common cause of cancer-related death. Despite significant progress in the diagnosis and treatment of this tumor, patients who relapse after radical treatments inevitably develop metastatic disease. Patient stratification is therefore key in this type of cancer, and there is an urgent need for prognostic biomarkers that can define patients’ risk of cancer-related death. In the last 10 years, multiple prognostic factors have been identified and studied. Here, we review the literature available and discuss the most common aberrant genomic pathways in metastatic castration-resistant prostate cancer shown to have a prognostic relevance in this setting. Prostate cancer (PC) is the second most common cancer for incidence and the fifth most frequent cause of cancer-related mortality in men worldwide, while in Western countries it represents the second cause of death for cancer in males ,2. The iProstate cancer is a highly heterogeneous disease, in both clinical and molecular aspects. PC may range from slowly progressing to very aggressive disease, mostly due to the development of resistance to treatment. Although advanced PC typically starts with a strong dependency on androgens, resulting in initial responses to androgen deprivation treatments, most patients progress to castration-resistant disease (CRPC) . CRPC stTP53 mutations, and RB1 loss [In addition to the different possible clinical presentations, intra-patient tumor heterogeneity and clonal evolution must be taken into account and represent a challenge in the management of patients. Several recurrent molecular pathways have been identified in the metastatic CRPC (mCRPC) molecular landscape, resulting in resistance to treatments and tumor progression, and impacting patients’ survival. These include androgen receptor (AR) aberrations, PTEN loss, DNA repair gene deletions, RB1 loss ,7.Androgens are involved in the normal development of the prostate gland, but also in prostate carcinogenesis. In normal prostate tissue, differentiation is promoted, while during the progression of PC, proliferation gradually overcomes differentiation. AR contributes to the control of the balance between cell proliferation and differentiation ,9,10,11.Androgen signaling inhibition, either through androgen deprivation or AR activity block, is therefore the mainstay of PC treatment. In the last decade, novel hormonal drugs have been approved in castration-sensitive prostate cancer (CSPC) and/or mCRPC thanks to their mechanisms of action  .AR gene mutations, AR splice-variants, an AR gene amplification; also, the presence of AR co-regulators may contribute to resistance to treatments [Indeed, even when the disease turns castration-resistant, AR remains an important molecular driver ,13,14,15eatments ,17. Theseatments .AR gene amplifications (or gain) [AR gene amplifications, AR antagonists  may be unable to effectively antagonize AR proteins. Moreover, since PC is able to synthetize androgens, even when androgen concentrations are under castrate levels, AR signaling still remains active. Through these mechanisms, AR amplifications and enhanced AR signaling result in resistance to first- and second-generation antiandrogens [The vast majority of patients with AR pathway alterations present or gain) . The effndrogens ,20.AR point mutations [AR T878A enables the activation of the receptor by progesterone and L702H the activation by glucocorticoids such as prednisone, which is given together with abiraterone and docetaxel. Other mutations frequently described include V715M, V730M, and H875Y [Other AR alterations such as utations  can alsond H875Y .AR amplifications [Secondary AR alterations, usually emerging after treatment with abiraterone or enzalutamide, are represented by AR splice variants (AR-Vs). AR-V7 is the most frequent variant, conferring a constitutive activation resulting in enhanced transcription, cell proliferation and DNA repair. Thus, AR-Vs are related to treatment resistance and poor outcomes ,23,24. Hications .Finally, the presence of AR co-regulators represents another possible mechanism of resistance to treatments. Several co-activators may interact with AR and enhance transcription. Among them, the activity of the members of the p160 steroid receptor coactivator (SRC) family, the histone acetyltransferase CBP/p300, and the pioneer factor forkhead box A1 (FOXA1) have been described to be associated with treatment resistance and worse prognosis ,26,27.p = 0.045) in taxane-treated patients. Using the same automated immunofluorescent assay of CTC, a multicenter study also confirmed the predictive value of AR-V7 positivity in 142 mCRPC patients, showing a benefit in OS for AR-positive patients treated with chemotherapy compared to those treated with AR-directed agents [All these alterations involving the AR pathway can be studied using liquid biopsies. For example, given the heterogeneity of AR-V7 expression at different metastatic sites , its stud agents .p = 0.23) and shorter median PFS  compared to the AR-V7-negative patients.Interestingly, in the TAXNERGY study , using a novel digital droplet PCR assay, 67% of the 54 docetaxel/ or cabazitaxel treated patients had AR-V7-positive CTCs . These pPreliminary findings were also reported in 15 AR-V7-positive patients treated with ipilimumab and nivolumab showing lack of efficacy in this subset of patients, with only two patients (13%) achieving a PSA response .AR gene aberrations that are infrequent in the early stages, but common in the CRPC setting. Therefore, NGS- and PCR-based studies have been particularly focused on AR copy number in advanced disease. Romanel et al. [AR gain and point mutations to be mutually exclusive. While AR copy number was not different during treatment, the emergence of T878A or L702H AR amino acid changes was reported in 13% of samples upon progression. There was also an association between plasma AR aberrations and worse OS  and PFS . Subsequently, a multicenter biomarker study assessed the role of plasma AR also in chemotherapy-naïve patients [AR gain, while 11% post-docetaxel but no chemotherapy-naïve abiraterone-treated patients showed AR point mutations. Chemotherapy-naïve and post-docetaxel AR-gained patients demonstrated a poor survival . Patients with AR mutations also showed a significantly worse OS . In the validation cohort, 11 (12%) patients had AR gain. AR-gained patients had a reduced biochemical PFS , radiographic PFS , OS .Further circulating biomarkers in CRPC are represented by l et al.  analyzedpatients . There wAR copy number has also been investigated in CRPC patients treated with taxanes [AR and outcomes in mCRPC, reporting only a significant worse OS in AR-gained patients . Moreover, the same authors incorporated updated data from their prior study [AR and treatment type in the abiraterone/enzalutamide-treated group (73 patients), after and in 115 first-line docetaxel patients. The findings from this study would suggest that AR-normal men could benefit from hormonal treatment in the first-line therapy group, while plasma AR-gained patients had a better response to docetaxel. Similarly, in a subsequent work with mCRPC patients receiving second-line cabazitaxel therapy, a significant treatment interaction between plasma AR and cabazitaxel vs. AR-directed therapies was observed for OS (p = 0.041). An exploratory analysis showed AR-gained patients treated with an initially reduced dose of cabazitaxel presented a meaningfully worse OS/PFS and would probably need a standard initial dose of cabazitaxel. This study suggests that plasma AR may improve clinical decision-making in choosing between adapted and standard regimen of taxanes [As previous findings have suggested that the detection of AR-V7 in CTCs could predict a response to AR-signaling inhibitors versus taxane chemotherapy, recently, the role of cell-free  taxanes ,39. A bior study , and intAR gene status has been also explored in a phase 2 clinical trial (NCT03454750) of mCRPC patients treated with 177Lu–PSMA–617 [AR amplification and 5 of 25 (20%) with AR normal (p = 0.0002)). The OR for patients with early disease progression and AR gain was 16.00, 95% CI 3.23–79.27, p = 0.0007. These preliminary data suggest that plasma AR status assessed by ddPCR could be helpful to identify resistance to 177Lu–PSMA–617 in mCRPC patients.Recently, plasma PSMA–617 . Early pAR mutations, e.g., F877L and T878A, responses to novel AR-directed therapies, such as apalutamide [AR mutations was so low that no conclusions could be drawn.Recent studies determined the impact of lutamide  and darolutamide . The phalutamide  used theAR copy number gain and an enrichment for AR binding sequences and hypomethylation of these segments. Moreover, plasma methylome analysis could accurately estimate tumor fraction. All these findings permitted to identify patients with a more aggressive clinical course based on this methylation pattern and so a molecular prognostic stratification of mCRPC individuals.A recent paper  uncovereThe role of plasma DNA analysis has been recently investigated in combination with functional imaging and other routinely obtained circulating biomarkers in order to improve prognostication of mCRPC in patients . This woPTEN, mainly through the loss of its locus on 10q23.31, is the most common molecular aberration involving this pathway occurring in approximately 40% of CRPCs [PIK3CA, PIK3CB, PIK3R1, PIK3R3 and AKT1 [Alterations in phosphatase and tensin homolog (PTEN) and activation of the kinases PI3K and AKT, downstream of its axis, are common in primary PC and are enriched in mCRPCs ,45. Genoof CRPCs ,7,45. Hoof CRPCs , as welland AKT1 . All theand AKT1 . The PTEand AKT1 ,49, and and AKT1 ,51,52,53p = 0.004), but also with shorter median duration on abiraterone . More importantly, PTEN status was consistent between matched CSPC and CRPC tumor biopsies in nearly 90% of cases [PI3K and AKT alterations were identified in 11% of 418 prostate cancer tissue samples, of these, 26/418 (6%) were pathogenic pathway activating mutations. These mutations were associated with a shorter OS  and duration of abiraterone/enzalutamide  [Ferraldeschi et al. analyzed the PTEN status of 144 patients who had received abiraterone post-docetaxel. In this retrospective study, the loss of PTEN expression was not only associated with shorter median OS  .Altogether, these data highlight the relevance of the aberrations in the PTEN pathway as prognostic factor in mCRPC. However, more recently, these alterations have been studied as biomarkers of response to the combination of new generation hormonal treatments and AKT inhibitors such as ipatasertib and capivasertib ,56.Genomic instability is a common denominator of many different cancers. This instability derives from the high rate of cell division that is responsible for the fast accumulation of genomic aberrations . TherefoBRCA2), a key member of the HR and Fanconi anemia complex, is the most commonly mutated DDR gene in prostate cancer. In the metastatic setting, germline BRCA2 (gBRCA) mutations have a prevalence of 3–6%, while somatic mutations and homozygous deletions account for ~20% of metastatic CRPC (mCRPC) cases [BRCA1, PALB2 and RAD51, are present in <1% of mCRPC. The prognostic role of germline and somatic mutations in HR genes, and specifically of BRCA2 alterations, has been investigated in detail in recent years. BRCA2 mutation carriers have a 5-year prostate cancer-specific survival (CSS) rate of ~50%, progressing rapidly from localized PC to mCRPC [gBRCA2 patients were found to have a shorter CSS compared to non-carriers  [BRCA2 mutation-carriers and 8,463 non-carriers confirmed that carrying a BRCA2 alteration correlates with a reduced CSS and OS (Hazard Ratios (HRs) 2.53 vs. 2.21 respectively, p < 0.001). The results also demonstrated that BRCA2-mutation carriers were characterized by a higher Gleason Score (GS) (>7), TNM stage  at diagnosis [Breast cancer-associated gene2 (C) cases ,19,60. Oto mCRPC ,62,63,64= 0.027) . A receniagnosis . The preiagnosis ,68,69; niagnosis ,71.ATM  is a member of the PI3 family of serine-threonine kinases and functions as a DNA damage sensor [BRCA1 or BRCA2 biallelic loss [e sensor . Therefoe sensor . Its alte sensor ,60, but lic loss ,75. Its lic loss .CDK12 are associated with a genome-wide focal tandem duplication (FTD) signature [CK12 mutant mCRPC, which are also characterized by a high Gleason score (>8) and a tumor immune infiltrate, potentially immunosuppressive, comprised of CD4+FOXP3- cells [CDK12 among the putative biomarkers of response to novel immunotherapies in mCRPC [CDK12 is reportedly involved in regulating expression of several HR genes ; somaticignature . Multipl3- cells . The FTDin mCRPC .MSH2 and MSH6, the two main players of the MMR system, involve noncoding regions detectable only by whole-genome studies. MMR-defective status (dMMR) was associated with poor OS compared to MMR-proficient tumors , increased T-cell infiltration and elevated PD-L1 protein expression in a cohort of 124 mCRPC biopsies from a single institution [The mismatch repair (MMR) system is a post-replicative, high-fidelity, single-strand repair mechanism that recognizes and reverses DNA base mismatches and insertion/deletion loops, compromised MMR results in microsatellite instability, and a hypermutator phenotype that has been associated with chemotherapy resistance but immunotherapy sensitivity . The pretitution . Transcrtitution .BRCA2 losses often co-occur with the retinoblastoma (RB1) ones [RB1 is a tumor suppressor that also plays a relevant role in mCRPC. Comprehensive genomic, transcriptomic and histologic analysis of 429 patients with mCRPC linked with longitudinal clinical outcomes identified RB1 genomic alteration as a potent predictor of poor outcome especially when associated with tumor protein p53 (TP53) alterations [RB1 and TP53 aberrations was also confirmed by another large retrospective study in which 470 treatment-naïve prostate cancer diagnostic biopsies and 61 matched mCRPC biopsies were analyzed using targeted and low-pass whole-genome sequencing. Moreover, this study demonstrated that RB1 losses along with TP53 and AR aberrations were enriched in later stages [Considering their localization on chromosome 13, B1) ones . RB1 is erations . TP53 plerations  and reprerations . The pror stages .We explored the relevance of prognostic biomarkers in mCRPC; however, in the last few years, some studies have proposed stratifying patients with localized PC based on molecular biomarkers. Traditionally, Gleason score, clinical staging and PSA at diagnosis have helped classifying localized PCs as being at low, intermediate, and high risk of relapse . RecentlUsing RNA analysis on formalin fixed paraffin embedded clinical samples, studying biological pathways with a known role in prostate tumorigenesis , these classifiers  offer scores that, integrating other clinical–pathological features, might help identifying patients with localized disease but with worse outcomes. Potentially, these tools could help clinicians in deciding which patients would benefit from surveillance vs. active treatment, or a salvage approach after a radical treatment ,90,91,92However, tissue-based molecular testing is dependent on tissue preservation and influenced by the heterogeneity of the disease . MoreoveHere, we presented the most common genomic pathway aberrations in prostate cancer that have an impact on patient outcomes. All of these studies and data are relevant to understanding the importance of genomic studies in the metastatic setting in order to better evaluate patient prognosis. Since some of these biomarkers can be used as either therapeutic targets or as biomarkers of response to novel agents and standard treatments in mCRPC, they also have emerging potential in targeted therapy and therapy response prediction .