Differential phosphorylation of Ca2+-permeable channel CNGC20 modulates calcium-mediated freezing tolerance in Arabidopsis.

Plants respond to cold stress at multiple levels, including increasing cytosolic calcium (Ca2+) influx and triggering the expression of cold-responsive genes. Here we show that the Ca2+-permeable channel CYCLIC NUCLEOTIDE GATED CHANNEL20 (CNGC20) positively regulates freezing tolerance in Arabidopsis (Arabidopsis thaliana) by mediating cold-induced Ca2+ influx. Moreover, we demonstrate that the leucine-rich repeat receptor-like kinase PLANT PEPTIDE CONTAINING SULFATED TYROSINE1 RECEPTOR (PSY1R) is activated by cold, phosphorylating and enhancing the activity of CNGC20. The psy1r mutant exhibited decreased cold-evoked Ca2+ influx and freezing tolerance. Conversely, COLD-RESPONSIVE PROTEIN KINASE1 (CRPK1), a protein kinase that negatively regulates cold signaling, phosphorylates and facilitates the degradation of CNGC20 under prolonged periods of cold treatment, thereby attenuating freezing tolerance. This study thus identifies PSY1R and CRPK1 kinases that regulate CNGC20 activity and stability, respectively, thereby antagonistically modulating freezing tolerance in plants.

Root-knot nematodes produce functional mimics of tyrosine-sulfated plant peptides.

Root-knot nematodes (Meloidogyne spp.) are highly evolved obligate parasites threatening global food security. These parasites have a remarkable ability to establish elaborate feeding sites in roots, which are their only source of nutrients throughout their life cycle. A wide range of nematode effectors have been implicated in modulation of host pathways for defense suppression and/or feeding site development. Plants produce a diverse array of peptide hormones including PLANT PEPTIDE CONTAINING SULFATED TYROSINE (PSY)-family peptides, which promote root growth via cell expansion and proliferation. A sulfated PSY-like peptide RaxX (required for activation of XA21 mediated immunity X) produced by the biotrophic bacterial pathogen (Xanthomonas oryzae pv. oryzae) has been previously shown to contribute to bacterial virulence. Here, we report the identification of genes from root-knot nematodes predicted to encode PSY-like peptides (MigPSYs) with high sequence similarity to both bacterial RaxX and plant PSYs. Synthetic sulfated peptides corresponding to predicted MigPSYs stimulate root growth in Arabidopsis. MigPSY transcript levels are highest early in the infection cycle. Downregulation of MigPSY gene expression reduces root galling and egg production, suggesting that the MigPSYs serve as nematode virulence factors. Together, these results indicate that nematodes and bacteria exploit similar sulfated peptides to hijack plant developmental signaling pathways to facilitate parasitism.

Potato Solanum tuberosum L. Phytoene Synthase Genes (StPSY1, StPSY2, and StPSY3) Are Involved in the Plant Response to Cold Stress.

The structure and phylogeny of the Solanum tuberosum L. phytoene synthase genes StPSY1, StPSY2, and StPSY3 were characterized. Their expression was studied in potato seedlings exposed to cold stress in the dark phase of the diurnal cycle to simulate night cooling. All of the three genes were activated as the temperature decreased, and the greatest response was observed for StPSY1. StPSY3 was for the first time shown to respond to cold stress and photoperiod. A search for cis-regulatory elements was carried out in the promoter regions and 5'-UTRs of the StPSY genes, and the regulation of all three genes proved associated with the response to light. A high level of cold-induced activation of StPSY1 was tentatively attributed to the presence of cis elements associated with sensitivity to cold and ABA.

Community Mental Health Services for American Indians and Alaska Natives: Reconciling Evidence-Based Practice and Alter-Native Psy-ence.

This review updates and extends Gone & Trimble's (2012) prior review of American Indian (AI) and Alaska Native (AN) mental health. First, it defines AI/AN populations in the USA, with an explanation of the importance of political citizenship in semisovereign Tribal Nations as primary for categorizing this population. Second, it presents an updated summary of what is known about AI/AN mental health, with careful notation of recurrent findings concerning community inequities in addiction, trauma, and suicide. Third, this article reviews key literature about AI/AN community mental health services appearing since 2010, including six randomized controlled trials of recognizable mental health treatments. Finally, it reimagines the AI/AN mental health enterprise in response to an "alter-Native psy-ence," which recasts prevalent mental health conditions as postcolonial pathologies and harnesses postcolonial meaning-making through Indigenized therapeutic interventions. Ultimately, AI/AN Tribal Nations must determine for themselves how to adopt, adapt, integrate, or refuse specific mental health treatments and services for wider community benefit.

Psychosocial issues among paediatric health-care workers posted in intensive care unit during COVID-19 pandemic: A questionnaire-based survey (Psy-Co-19 survey).

AIM: To understand the moral distress experienced by health-care workers (HCWs) in the COVID paediatric intensive care unit (PICU). We also aimed to assess the psychological well-being and the coping mechanisms used by HCWs. METHODS: A prospective observational cross-sectional study was conducted from July to September 2021, involving all HCWs who worked in the COVID PICU. Moral distress using Moral Distress for Health-care Professionals (MMD-HPs) scale, psychological well-being using Trauma Screening Questionnaire (TSQ) and coping strategies adopted by HCWs using Brief-COPE (Coping Orientation to Problems Experienced) were measured. RESULTS: One hundred and eighty-four HCW data were examined. The most common causes of moral distress among HCWs were compromised patient care caused by a lack of resources and caring for more patients than they could safely handle. Moral distress was the same regardless of the HCWs' job profile, marital status, number of children or age. The TSQ revealed psychological stress in 23.3% of HCWs with Post-traumatic Stress Disorder, significantly higher in HCWs under the age of 30 and without children. Few HCWs turned to substance use, self-blame or denial as coping mechanisms; instead, acceptance, self-distraction and emotional support were the most frequently used. CONCLUSION: The most common reasons for moral and psychological distress perceived by participants were insufficient staff and organisational support. Younger HCWs and those without children experienced higher levels of psychological distress. HCWs' typical coping mechanisms are constructive, such as seeking help and support from others, reframing situations and meditation. Health-care administrators must develop a framework to assist HCWs in dealing with such serious issues.

Transcript expression level analysis of phytoene synthase and phytoene desaturase associated with ß-carotene content in selected Kenyan Bitter melon.

BACKGROUND: Bitter melon (Momordica charantia L.) is a widely cultivated food and medicinal plant native to the world's subtropics and tropics. Constraints affecting cultivation of Bitter melon affect productivity of ß-carotene. Knowing the mechanism that controls the transcription of the ß-carotene biosynthesis genes in Bitter melon will be of great value in improving the yield of this important metabolite. METHODS AND RESULTS: The expressions of ß-carotene biosynthetic genes such as Phytoene Desaturase (PDS) and Phytoene Synthase (PSY) were evaluated in Bitter melon accessions 'GBK027049', 'NS1026', 'Mahy-ventura', '453B' and 'Sibuka532'. Transcript expression level analysis of PSY and PDS, and amount of ß-carotene in leaf, stem, and fruit, were determined using quantitative polymerase chain reaction and high-performance liquid chromatography (HPLC). Root transcript expression was used as a negative control for determining relative fold change in other tissues. Expression of PSY in fruit (6 to 27-fold compared to the control) was higher than in the other organs for all accessions. This was also the case of PDS expression (10 to 29-fold compared to the control). Leaves had the highest ß-carotene concentration (17.92-45.35 µg∙g-1); there was no difference between stems (5.67-12.75 µg∙g-1) and fruit (6.18-12.53 µg∙g-1). The highest ß-carotene content was in accessions 'GBK027049' (12.53-45.35 µg∙g-1) and '453B' (6.18-32.09 µg∙g-1). The PSY and PDS expressions were positively correlated with amount of ß-carotene in leaves, stems, and fruits. CONCLUSION: Bitter melon leaves, especially those of 'GBK027049' and '453B' accessions, are an alternative to alleviate the ß-carotene deficiencies in the world and especially in Africa.

Baseline depressive symptoms, personal control, and concern moderate the effects of preoperative psychological interventions: the randomized controlled PSY-HEART trial.

This study examined whether baseline (3-14 days pre-surgery) levels of (i) depressive or (ii) anxiety symptoms and (iii) illness beliefs moderate the effects of additional preoperative interventions before coronary artery bypass graft surgery on (i) depressive or (ii) anxiety symptoms and (iii) illness beliefs 1 day before surgery, 1 week and 6 months after surgery. In the PSY-HEART trial, 115 patients were assessed. They were randomized into one of three groups: 1. receiving standard medical care only (SMC), additional psychological interventions: 2. aiming to optimize patients' expectations (EXPECT), or 3. focusing on emotional support. Patients with a higher baseline level of depressive symptoms receiving a preoperative psychological intervention indicated lower depressive symptoms 6 months after surgery compared to SMC. EXPECT increased personal control and concern levels in patients with low baseline personal control/concern 1 day before surgery. Brief preoperative psychological interventions can improve psychological outcomes in heart surgery patients. Baseline status may moderate these effects. The study has been approved by the medical ethics committee of the Philipps University of Marburg and has been pre-registered at www.clinicaltrials.gov (NCT01407055) on August 1, 2011.

Anti-Cancer Effects of a New Herbal Medicine PSY by Inhibiting the STAT3 Signaling Pathway in Colorectal Cancer Cells and Its Phytochemical Analysis.

Colorectal cancer (CRC) is an inflammation-associated common cancer worldwide. Paejang-san and Mori Cortex Radicis have been traditionally used for treating intestinal inflammatory diseases in Korea and China. In the present study, we developed a new herbal formula as an alternative to CRC treatments, which is composed of two main components of Paejangsan (Patriniae Radix (Paejang in Korean) and Coix Seed (Yiyiin in Korean)), and Mori Cortex Radicis (Sangbekpi in Korean) based on the addition and subtraction theory in traditional medicine, hence the name PSY, and explored the potential therapeutic effects of the new formula PSY in human CRC cells by analyzing viability, cell cycle and apoptosis. We found that PSY ethanol extract (EtOH-Ex), but not water extract, significantly suppressed the viability of human CRC cells, and synergistically decreased the cell proliferation compared to each treatment of Patriniae Radix and Coix Seed extract (PY) or Mori Cortex Radicis extract (S), suggesting the combination of PY and S in a 10-to-3 ratio for the formula PSY. PSY EtOH-Ex in the combination ratio reduced cell viability but induced cell cycle arrest at the G2/M and sub-G1 phases as well as apoptosis in CRC cells. In addition, the experimental results of Western blotting, immunofluorescence staining and reporter assays showed that PSY also inhibited STAT3 by reducing its phosphorylation and nuclear localization, which resulted in lowering STAT3-mediated transcriptional activation. In addition, PSY regulated upstream signaling molecules of STAT3 by inactivating JAK2 and Src and increasing SHP1. Moreover, the chemical profiles of PSY from UPLC-ESI-QTOF MS/MS analysis revealed 38 phytochemicals, including seven organic acids, eight iridoids, two lignans, twelve prenylflavonoids, eight fatty acids, and one carbohydrate. Furthermore, 21 potentially bioactive compounds were highly enriched in the PSY EtOH-Ex compared to the water extract. Together, these results indicate that PSY suppresses the proliferation of CRC cells by inhibiting the STAT3 signaling pathway, suggesting PSY as a potential therapeutic agent for treating CRC and 21 EtOH-Ex-enriched phytochemicals as anti-cancer drug candidates which may act by inhibiting STAT3.

Idiopathic granulomatous mastitis.

Idiopathic granulomatous mastitis mainly affects women of childbearing age, mostly several months to years after breastfeeding. The etiopathogenesis is unknown to this day. Because clinical manifestations can mimic infectious mastitis or inflammatory carcinoma, the dia-gnostic process is often prolonged. The correct dia-gnosis is determined only on the basis of histopathological findings. To this day, there is no uniform consensus on the treatment of idiopathic granulomatous mastitis. The chronic nature of the disease as well as the tendency to recurrences despite the benign nature of the disease affects the quality of life of patients. Case reports from the Gynecoly and Obstetrics Clinic in University Hospital in Trenčín also point to an adverse effect on womens lives.

Evidence for multiple receptors mediating RALF-triggered Ca2+ signaling and proton pump inhibition.

Acidification of the apoplastic space facilitates cell wall loosening and is therefore a key step in cell expansion. PSY1 is a growth-promoting secreted tyrosine-sulfated glycopeptide whose receptor directly phosphorylates and activates the plasma membrane H+ -ATPase, which results in acidification and initiates cellular expansion. Although the mechanism is not clear, the Rapid Alkalinization Factor (RALF) family of small, secreted peptides inhibits the plasma membrane H+ -ATPase, leading to alkalinization of the apoplastic space and reduced growth. Here we show that treating Arabidopsis thaliana roots with PSY1 induced the transcription of genes encoding the RALF peptides RALF33 and RALFL36. A rapid burst of intracellular Ca2+ preceded apoplastic alkalinization in roots triggered by RALFs, with peptide-specific signatures. Ca2+ channel blockers abolished RALF-induced alkalinization, indicating that the Ca2+ signal is an obligatory part of the response and that it precedes alkalinization. As expected, fer mutants deficient in the RALF receptor FERONIA did not respond to RALF33. However, we detected both Ca2+ and H+ signatures in fer mutants upon treatment with RALFL36. Our results suggest that different RALF peptides induce extracellular alkalinization by distinct mechanisms that may involve different receptors.

IL-8 and CRP moderate the effects of preoperative psychological interventions on postoperative long-term outcomes 6 months after CABG surgery - The randomized controlled PSY-HEART trial.

INTRODUCTION: Inflammation has been related to several somatic and psychological disorders and may moderate effects of psychological interventions. In the PSY-HEART trial patients benefitted from preoperative psychological interventions before undergoing coronary artery bypass graft surgery (CABG) and, if necessary, concomitant valvular surgery, compared to standard medical care. In this study we examined whether patients' baseline inflammatory status moderated the intervention effects. MATERIAL AND METHODS: In a prospective three-arm randomized clinical trial with 6-months follow-up, 124 patients scheduled for CABG surgery alone or concomitant with valvular surgery were randomized to (i) standard medical care only (SMC) or two preoperative psychological interventions: (ii) CBT-based optimizing expectations (EXPECT) and an (iii) an active control group focusing on emotional support (SUPPORT). Available baseline CRP- (n = 79), IL-6- (n = 78), IL-8- (n = 78) and TNF-alpha-(n = 80) parameters were considered as potential moderators (CRP as a categorical and continuous moderator). Linear mixed model analyses were calculated to test whether baseline inflammatory levels moderated intervention effects on disability, mental and physical quality of life at 6 months after surgery. RESULTS: IL-8 moderated intervention effects on patients' disability and categorical CRP moderated intervention effects on mental quality of life. Follow-up tests indicated that EXPECT (and in part SUPPORT) led to lower postoperative disability and higher mental quality of life compared to SMC in patients with low baseline inflammatory markers. EXPECT indicated higher mental quality of life compared to SUPPORT in the high CRP subgroup. Patients in the SMC group had higher mental quality of life in the high CRP subgroup compared to the low CRP subgroup. CONCLUSION: Especially for patients with a lower inflammatory baseline status preoperative psychological interventions might be helpful to optimize long-term CABG surgery outcomes.

Working toward the ideal situation: A pragmatic Epi-Psy approach for the diagnosis and treatment of psychogenic nonepileptic seizures.

Medical international guidelines recommend regular psychiatric consultations for patients with epilepsy, in order to detect comorbidities. However, there is a lack of guidance about PNES that constitute both a differential diagnosis and a comorbidity of epilepsy. While waiting for the ideal collaboration between neurologists and psychiatrists, we develop a pragmatic approach. Wrong diagnosis between epilepsy and Psychogenic nonepileptic seizures (PNES) is frequent and may lead to iatrogenic consequences for patients. To limit the risk of misdiagnosis, psychiatrists and neurologists should collaborate and be more trained about epilepsy, PNES, and their comorbidities. We illustrate the aim of this collaboration through the case of a patient, initially diagnosed with epilepsy, then with PNES only and finally with comorbid epilepsy and PNES. The correct final diagnosis would not have been performed without the collaboration of psychiatrists and neurologists, trained in "Epi-Psy" approach.

Disparity of the carotenoids antioxidant properties of wild-type and D-PSY-transgenic Dunaliella parva strains under three environmental stresses.

Two strains of the halophilic alga Dunaliella parva, a wild type (WT) and a transgenic strain (D-PSY) containing an exogenous phytoene synthase gene (PSY), were used to investigate the growth, carotenoid accumulation, and carotenoid antioxidant properties under nitrogen starvation, cobalt and biochar treatments. D-PSY had higher carotenoid content (1.8 times) compared to the WT. The applied stressors stimulated the carotenoid content of both WT and D-PSY especially. The carotenoids were assayed for the potential antioxidant activities by five different assays. Generally, the antioxidant activities of D-PSY carotenoids were superior to that of WT. The biochar and nitrogen treatments generally enhanced the antioxidant activities of the carotenoids, whereas cobalt came third in this respect. The D-PSY transgenic algal strain has both high carotenoids content and antioxidant properties which enhanced under the relatively lower concentrations of the applied stressors. The results have shown to lead to an accurate application of the transgenic alga as a source of potent antioxidant compounds. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01077-0.

Effects of prebiotic dietary fibers and probiotics on human health: With special focus on recent advancement in their encapsulated formulations.

BACKGROUND: Dietary fibers (DFs) are known as potential formulations in human health due to their beneficial effects in control of life-threatening chronic diseases including cardiovascular disease (CVD), diabetes mellitus, obesity and cancer. In recent decades scientists around the globe have shown tremendous interest to evaluate the interplay between DFs and gastrointestinal (GIT) microbiota. Evidences from various epidemiological and clinical trials have revealed that DFs modulate formation and metabolic activities of the microbial communities residing in the human GIT which in turn play significant roles in maintaining health and well-being. Furthermore, interestingly, a rapidly growing literature indicates success of DFs being prebiotics in immunomodulation, namely the stimulation of innate, cellular and humoral immune response, which could also be linked with their significant roles in modulation of the probiotics (live beneficial microorganisms). SCOPE AND APPROACH: The main focus of the current review is to expressively highlight the importance of DFs being prebiotics in human health in association with their influence on gut microbiota. Now in order to significantly achieve the promising health benefits from these prebiotics, it is aimed to develop novel formulations to enhance and scale up their efficacy. Therefore, finally, herein unlike previously published articles, we highlighted different kinds of prebiotic and probiotic formulations which are being regarded as hot research topics among the scientific community now a days. CONCLUSION: The information in this article will specifically provide a platform for the development of novel functional foods the demands for which has risen drastically in recent years.

Co-constructing a decolonising praxis in academia through dialogues and pedagogical experiences between the UK and Ecuador.

As demanded by Argentinian cartoonist Quino's (2014) character Libertad (meaning freedom), the Latin American tradition of Liberation Psychology has explicitly called for a decolonising praxis since the 1970s. Such call has implications for training courses in psychology and other fields related to health and wellbeing, such as psychiatrists or social workers. However, there are various challenges in translating a decolonial rhetoric into concrete practice within the classroom, research and practice placements. This paper presents an account of experience and dialogues between two liberatory and critical community psychology educators in universities from the UK and Ecuador. Based on an international participatory roundtable on decolonisation and a pedagogical exchange between their students, the authors discuss the limitations and possibilities of engaging in a 'new praxis' that is both decolonial and liberatory.

Dysregulated autophagy as a new aspect of the molecular pathogenesis of Krabbe disease.

Krabbe disease (KD) is a childhood leukodystrophy with no cure currently available. KD is due to a deficiency of a lysosomal enzyme called galactosyl-ceramidase (GALC) and is characterized by the accumulation in the nervous system of the sphingolipid psychosine (PSY), whose cytotoxic molecular mechanism is not fully known yet. Here, we study the expression of some fundamental autophagy markers (LC3, p62, and Beclin-1) in a KD murine model [the twitcher (TWI) mouse] by immunohistochemistry and Western blot. Moreover, the autophagy molecular process is also shown in primary fibroblasts from TWI and WT mice, with and without PSY treatment. Data demonstrate that large p62 cytoplasmic aggregates are present in the brain of both early and late symptomatic TWI mice. p62 expression is also upregulated in TWI sciatic nerves compared to that measured for WT nerves. In vitro data suggest that this effect might not be fully PSY-driven. Finally, we investigate in vitro the capability of autophagy inducers (Rapamycin, RAP and Resveratrol, RESV) to reinstate the WT phenotype in TWI cells. We show that RAP administration can partially restore the autophagy markers levels, while RESV cannot, indicating a line along which new therapeutic approaches can be developed.

A tomato B-box protein SlBBX20 modulates carotenoid biosynthesis by directly activating PHYTOENE SYNTHASE 1, and is targeted for 26S proteasome-mediated degradation.

Carotenoids play important roles in many biological processes, such as light harvesting, photoprotection and visual attraction in plants. However, the regulation of carotenoid biosynthesis is still not fully understood. Here, we demonstrate that SlBBX20, a B-box (BBX) zinc-finger transcription factor, is a positive regulator of carotenoid accumulation in tomato (Solanum lycopersicum). Overexpression of SlBBX20 leads to dark green fruits and leaves and higher levels of carotenoids relative to the wild-type. Interactions between SlBBX20 and DE-ETIOLATED 1 (SlDET1) lead to the ubiquitination and 26S proteasome-mediated degradation of SlBBX20. Moreover, deficiencies in the components of the CUL4-DDB1-DET1 complex enhanced the stability of the SlBBX20 protein. Thus, we conclude that SlBBX20 is a substrate of the CUL4-DDB1-DET1 E3 ligase. SlBBX20 can activate the expression of PHYTOENE SYNTHASE 1, encoding a key enzyme in carotenoid biosynthesis, by directly binding to a G-box motif in its promoter, which results in the elevated levels of carotenoids in SlBBX20 overexpression lines. We identified a key regulator of carotenoid biosynthesis and demonstrated that the stability of SlBBX20 is regulated by ubiquitination. These findings provide us a new target for the genetic improvement of the nutritional quality of tomato fruit.

Co-production of farnesol and coenzyme Q10 from metabolically engineered Rhodobacter sphaeroides.

BACKGROUND: Farnesol is an acyclic sesquiterpene alcohol present in the essential oils of various plants in nature. It has been reported to be valuable in medical applications, such as alleviation of allergic asthma, gliosis, and edema as well as anti-cancerous and anti-inflammatory effects. Coenzyme Q10 (CoQ10), an essential cofactor in the aerobic respiratory electron transport chain, has attracted growing interest owing to its clinical benefits and important applications in the pharmaceutical, food, and health industries. In this work, co-production of (E,E)-farnesol (FOH) and CoQ10 was achieved by combining 3 different exogenous terpenes or sesquiterpene synthase with the RNA interference of psy (responsible for phytoene synthesis in Rhodobacter sphaeroides GY-2). RESULTS: FOH production was significantly increased by overexpressing exogenous terpene synthase (TPS), phosphatidylglycerophosphatase B (PgpB), and sesquiterpene synthase (ATPS), as well as RNAi-mediated silencing of psy coding phytoene synthase (PSY) in R. sphaeroides strains. Rs-TPS, Rs-ATPS, and Rs-PgpB respectively produced 68.2%, 43.4%, and 21.9% higher FOH titers than that of the control strain. Interestingly, the CoQ10 production of these 3 recombinant R. sphaeroides strains was exactly opposite to that of FOH. However, CoQ10 production was almost unaffected in R. sphaeroides strains modified by psy RNA interference. The highest FOH production of 40.45 mg/L, which was twice as high as that of the control, was obtained from the TPS-PSYi strain, where the exogenous TPS was combined with the weakening of the phytoene synthesis pathway via psy RNA interference. CoQ10 production in TPS-PSYi, ATPS-PSYi, and PgpB-PSYi was decreased and lower than that of the control strain. CONCLUSIONS: The original flux that contributed to phytoene synthesis was effectively redirected to provide precursors toward FOH or CoQ10 synthesis via psy RNA interference, which led to weakened carotenoid synthesis. The improved flux that was originally involved in CoQ10 production and phytoene synthesis was redirected toward FOH synthesis via metabolic modification. This is the first reported instance of FOH and CoQ10 co-production in R. sphaeroides using a metabolic engineering strategy.

Low Plasma Cholinesterase Activity is Associated With Postoperative Delirium After Noncardiac Surgery in Elderly Patients: AProspective Observational Study.

BACKGROUND: Postoperative delirium (POD) commonly occurs in elderly patients after noncardiac surgery, resulting in increased morbidity and greater risk of death. However, its pathophysiology is currently unknown. Cholinergic dysfunction has been implicated in delirium pathophysiology, and low plasma cholinesterase activity has been reported as a risk marker of POD. OBJECTIVE: Therefore, the aim of this study was to investigate the link between plasma cholinesterase activity and POD in elderly Han Chinese patients after noncardiac surgery. METHOD: From January 2014 to January 2016, a cohort of 206 patients aged ≥ 60years who underwent noncardiac surgery and were transferred to the surgical intensive care unit were enrolled. POD was assessed using the Confusion Assessment Method for Intensive Care Unit. Clinical data including sex, age, general comorbidities, alcohol consumption, Acute Physiology and Chronic Health Evaluation (APACHE) II score, and type of surgery were recorded. Blood was drawn postoperatively to measure cholinesterase activity. Using multiple logistic regression analyses, the associations between cholinesterase activity and POD were examined, adjusting for potential confounding variables. RESULTS: Delirium incidence was 22.3%. POD was associated with cholinesterase activity, age, and APACHE II score. In multiple logistic regression analyses, lower acetylcholinesterase and butyrylcholinesterase activity were independent risk factors for POD. CONCLUSION: Plasma cholinesterase activity may be a candidate biomarker for POD after noncardiac surgery in the elderly Chinese Han population.

CRISPR/Cas9 editing of carotenoid genes in tomato.

CRISPR/Cas9 technology is rapidly spreading as genome editing system in crop breeding. The efficacy of CRISPR/Cas9 in tomato was tested on Psy1 and CrtR-b2, two key genes of carotenoid biosynthesis. Carotenoids are plant secondary metabolites that must be present in the diet of higher animals because they exert irreplaceable functions in important physiological processes. Psy1 and CrtR-b2 were chosen because their impairment is easily detectable as a change of fruit or flower color. Two CRISPR/Cas9 constructs were designed to target neighboring sequences on the first exon of each gene. Thirty-four out of forty-nine (69%) transformed plants showed the expected loss-of-function phenotypes due to the editing of both alleles of a locus. However, by including the seven plants edited only at one of the two homologs and showing a normal phenotype, the editing rate reaches the 84%. Although none chimeric phenotype was observed, the cloning of target region amplified fragments revealed that in the 40% of analyzed DNA samples were present more than two alleles. As concerning the type of mutation, it was possible to identify 34 new different alleles across the four transformation experiments. The sequence characterization of the CRISPR/Cas9-induced mutations showed that the most frequent repair errors were the insertion and the deletion of one base. The results of this study prove that the CRISPRCas9 system can be an efficient and quick method for the generation of useful mutations in tomato to be implemented in breeding programs.