TM Treatment Research Articles

DAILY WRITING PROGRAM OF ENGLISH DIARY TO IMPROVE THE STUDENTS’ ABILITY TO WRITE

This research was aimed to investigate diary writing activity to improve the students’ writing ability at SMA PGRI 2 Palembang. The question of this study was to determine about, was there any significant difference between the students were taught by using diary writing activity and those were not. This research was conducted in SMA PGRI 2 Palembang, for the second semester 2020-2021 school year. This research could conclude that the diary writing activity to improve the students’ writing ability have a significant impact on students’ writing performance, because based on the result of the test, the students who were taught recount text through writing diary got higher scores than those who were not. From the students’ treatment process, the students in experimental group could improve their writing through diary writing. The average student’s writing performance in experimental group before and after the use of diary writing is significantly improved strategy.

Potent and Orally Bioavailable BCL6 PROTAC TM Degraders Demonstrate Efficacy in Pre-Clinical Models of Diffuse Large B-Cell Lymphoma (DLBCL)

The BCL6 transcription repressor (B-cell lymphoma 6, BCL6) protein has been shown to be a key molecular driver of diffuse large B-cell lymphoma (DLBCL). Somatic mutations of the BCL6 gene include gross- or cryptic-chromosome translocations and point mutations that have been shown to result in the deregulation of BCL6 expression. These BCL6 abnormalities also contribute to a subgroup of high-risk (HR) aggressive double- and triple-hit (DH/TH) lymphomas with very poor outcomes. We have developed highly specific, potent and orally bioavailable BCL6 PROteolysis TArgeting Chimera (PROTAC TM) degraders that demonstrate potent in-vitro and in-vivo efficacy in multiple pre-clinical DLBCL models.Ten of 12 germinal center B-cell (GCB) and two of four activated B-cell (ABC) DLBCL cell lines show significant growth inhibition in-vitro with BCL6 PROTAC TM treatment, demonstrating a critical dependence on BCL6. This array of sensitivity across genetically variable cell lines suggests that BCL6-dependence is not just associated with BCL6-mutated DLBCLs. A more advanced BCL6 PROTAC TM, ARVN-71228, achieves >95% BCL6 D max in-vitro at a DC 50 of <1 nM in the OCI-Ly1 model following 24 hr treatment, degrading BCL6 equally well in the nuclear, chromatin-bound and cytosolic cell fractions. BCL6 degradation is associated with dose-dependent G1 cell cycle arrest and elevated apoptosis that increases over time (24 vs 72 hours). In head-to-head BCL6 degradation and growth inhibition studies using OCI-Ly1, the ARVN-71228 BCL6 PROTAC TM demonstrates superior activity compared to recently published BCL6-targeted degraders/inhibitors and heterobifunctional molecules.Importantly, medicinal chemistry efforts have resulted in the successful development of orally bioavailable BCL6 PROTAC TM degraders for in-vivo dosing. Time-course studies show >95% BCL6 loss within four hours which is maintained at 8-, 16- and 24-hours. Genes repressed by BCL6 such as BLIMP1 and PTPN6 are derepressed and show increased protein levels 24 hours post-dose. ARVN-71228 achieves regressions in the GCB OCI-Ly1 CDX model. Future studies plan to look at rational drug combinations with BCL6 PROTAC TM degraders to find collaborative or synergistic pathways to target, especially in the HR-DLBCL subtypes where there is a high unmet medical need. DisclosuresGough: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Sherman: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. DeCarr: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Eaton: Arvinas: Current Employment, Current equity holder in publicly-traded company. Milanovic: Arvinas: Current Employment, Current equity holder in publicly-traded company. Bookbinder: Arvinas: Current Employment, Current equity holder in publicly-traded company. Pizzano: Arvinas: Current Employment, Current equity holder in publicly-traded company. Altieri: Arvinas: Current Employment, Current equity holder in publicly-traded company. Corradi: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Xiao: Arvinas: Current Employment, Current equity holder in publicly-traded company. Gallego: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Soto: Arvinas: Current Employment, Current equity holder in publicly-traded company. Lingamaneni: Arvinas: Current Employment, Current equity holder in publicly-traded company. Chen: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Zhang: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Wang: Arvinas: Current Employment, Current equity holder in publicly-traded company. Dong: Arvinas: Current Employment, Current equity holder in publicly-traded company. Chirnomas: Arvinas: Current Employment, Current equity holder in publicly-traded company. Berlin: Arvinas: Current Employment, Current equity holder in publicly-traded company. Hornberger: Arvinas: Current Employment, Current equity holder in publicly-traded company. Snyder: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months. Taylor: Arvinas: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months.

6-BENZYLAMINOPURINE AND 2, 4-DICHLOROPHENOXYACETIC ACID EFFECT ON CALLUS GENESIS OF Brosimum alicastrum

Brosimum alicastrum seeds contain a high percentage of protein and essential amino acids that contribute to a proper nutrition. This tree is an alternative in the so-called crusade against hunger and its management does not involve the use of agrochemicals. The objective of this research was to evaluate the morphogenic response of foliar explants with two growth regulators, 6-Benzylaminopurine (BAP) and 2,4-Dichlorophenoxyacetic Acid (2,4-D) in the induction of calli. Results demonstrated that the combination of 1.5 mg L-1 BAP and 1 mg L-1 2,4-D (TM) and 1.5 mg L-1 BAP with 2 mg L-1 2,4-D (TN) favored callus growth in 100% of foliar explants. Calli were grown in a period of 20 d, in a culture room at 25 ± 4 °C, with 16 h of illumination. Under these conditions, calli remained in slow growth for four weeks. Those explants that generated callus were sub-cultured in fresh medium without activated charcoal. In TM and TN treatments, the multiplication of the cell mass was favored; in TN globular structures were formed. However, explants the same treatments TM and TN with activated charcoal, and under ambient 29 ± 4 °C conditions increased callus growth, but became friable at two weeks. Thus, TM and TN were the better treatments, but activated charcoal was determined to have a negative effect on callus growth.

Endoplasmic reticulum stress induces spatial memory deficits by activating GSK‐3 in Alzheimer’s disease

AbstractBackgroundEndoplasmic reticulum (ER) stress is involved in Alzheimer’s disease (AD), but the mechanism remains unclear.MethodTunicamycin (TM), a recognized ER stress inducer, was injected into the brain ventricle of Sprague‐Dawley (SD) rats to induce the unfolded protein response (UPR). Rats was divided into 3 groups for the brain lateral ventricle injections with DMSO, TM, or TM + SB, respectively. The behavioural test was conducted after 24 and 48 hours, respectively. Then the rats were sacrificed. Brain samples were collected for further protein analysis and specific stains.ResultTM‐induced UPR was convinced by enhanced phosphorylation of pancreatic ER kinase (PERK), inositol‐requiring enzyme‐1 (IRE‐1), and transcription factor‐6 (ATF‐6) activation. Behavioural test demonstrated UPR induced spatialmemory deficits and impairments of synaptic plasticity in the rats. After TM treatment, GSK‐3β was activated. Phosphorylation of cAMP response element binding protein at Ser129 (pS129‐CREB) was up‐regulated with an increased nuclear co‐localization of pY126‐GSK‐3β and pS129‐CREB. Simultaneous inhibition of GSK‐3β by hippocampal infusion of SB216763 (SB) attenuated TM‐induced UPR, synaptic plasticity, and spatial memory impairment with restoration of pS129‐CREB.ConclusionTM‐induced UPR causes AD‐like spatial memory deficits and synapse impairments by activating GSK‐3b/pS129‐CREB pathway. GSK‐3 inhibition might be a promising therapy in AD treatment.

Inositol-requiring enzyme 1 (IRE1) plays for AvrRpt2-triggered immunity and RIN4 cleavage in Arabidopsis under endoplasmic reticulum (ER) stress

Many stresses induce the accumulation of unfolded and misfolded proteins in the endoplasmic reticulum, a phenomenon known as ER stress. In response to ER stress, cells initiate a protective response, known as unfolded protein response (UPR), to maintain cellular homeostasis. The UPR sensor, inositol-requiring enzyme 1 (IRE1), catalyzes the cytoplasmic splicing of bZIP transcription factor-encoding mRNAs to activate the UPR signaling pathway. Recently, we reported that pretreatment of Arabidopsis thaliana plants with tunicamycin, an ER stress inducer, increased their susceptibility to bacterial pathogens; on the other hand, IRE1 deficient mutants were susceptible to Pseudomonas syringae pv. maculicola (Psm) and failed to induce salicylic acid (SA)-mediated systemic acquired resistance. However, the functional relationship of IRE1 with the pathogen and TM treatment remains unknown. In the present study, we showed that bacterial pathogen-associated molecular patterns (PAMPs) induced IRE1 expression; however, PAMP-triggered immunity (PTI) response such as callose deposition, PR1 protein accumulation, or Pst DC3000 hrcC growth was not altered in ire1 mutants. We observed that IRE1 enhanced plant immunity against the bacterial pathogen P. syringae pv. tomato DC3000 (Pst DC3000) under ER stress. Moreover, TM-pretreated ire1 mutants were more susceptible to the avirulent strain Pst DC3000 (AvrRpt2) and showed greater cell death than wild-type plants during effector-triggered immunity (ETI). Additionally, Pst DC3000 (AvrRpt2)-mediated RIN4 degradation was reduced in ire1 mutants under TM-induced ER stress. Collectively, our results reveal that IRE1 plays a pivotal role in the immune signaling pathway to activate plant immunity against virulent and avirulent bacterial strains under ER stress.

Abstract 6504: Field-in-field technique with intra-fractionally modulated junction shifts for craniospinal irradiation (CSI) planning with 3D-CRT

Abstract Purpose: To plan craniospinal irradiation with ‘‘field-in-field'' (FIF) homogenization technique in combination with daily, intra-fractional modulation of the field junctions, to minimize the possibility of spinal cord overdose. Photon-based techniques for craniospinal irradiation (CSI) may result in dose inhomogeneity within the treatment volume and usually require a weekly manual shift of the field junctions to minimize the possibility of spinal cord overdose. Nowadays field-in-field technique is used to feather out the dose inhomogeneity caused by multiple fields. We have started using this technique after acquiring advanced technology machines in recent years. Methods and Materials: 16 patients (2 adults, 14 children) treated with 3D-CRT for craniospinal irradiation were retrospectively chosen for this analysis. These patients were planned and treated during 2016-2017. Contouring of Brain and Spine Cord and organ at risk were already done and planning done on Eclipse TM Treatment Planning System (Varian). All of these patients were planned Lateral cranio-cervical fields and posterior spinal fields were planned using a forward-planned, FIF technique. Field junctions were automatically modulated and custom-weighted for maximal homogeneity within each treatment fraction. Dose volume histogram (DVH) was used for analysis of results. A corresponding plan without FIF technique was planned and maximum dose (in terms of volume of the PTV covering &amp;gt;110 % and &amp;gt;120 %) at the junction was noted for each patient with both plans and the readings were evaluated. Results: Plan inhomogeneity improved with FIF technique. Planning with daily-modulated junction shifts provided consistent dose delivery during each fraction of treatment across the junctions. Without FIF or modulated junctions, the thecal sac volume receiving 110% of the prescribed dose ranged from 12% to 25.8% (mean: 15%) and volume receiving 120% dose ranged from 0% to 8% (mean: 4%). Whereas with FIF and modulated junctions, the thecal sac volume receiving 110% of dose ranged from 0.59% to 3% and volume receiving 120% was 0%. Conclusion: Without FIF or modulated junctions, the thecal sac volume receiving 110% of the prescribed dose ranged from 12% to 25.8% (mean: 15%) and volume receiving 120% dose ranged from 0% to 8% (mean: 4%). Whereas with FIF and modulated junctions, the thecal sac volume receiving 110% of dose ranged from 0.59% to 3% and volume receiving 120% was 0%. Therefore it was concluded that Field-in-field technique improved dose conformality and reduced inhomogeneity. Citation Format: Shabbir Hussain A. Ali, Rahim Gohar, Hira Nazim, Jawaid Mallick. Field-in-field technique with intra-fractionally modulated junction shifts for craniospinal irradiation (CSI) planning with 3D-CRT [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6504.

PpAOX regulates ER stress tolerance in Physcomitrella patens

Severe environments disturb the folding or assembly of newly synthesized proteins, resulting in accumulation of misfolded or unfolded proteins in the endoplasmic reticulum (ER) as well as cytotoxic aggregation of abnormal proteins. Therefore, ER stress is evoked due to disturbed ER homeostasis. Alternative oxidase (AOX) plays an important role in coping with various abiotic stresses and plant growth. Our previous study has reported that PpAOX is involved in the regulation of salt tolerance in moss Physcomitrella patens (P. patens), but its biological functions in modulating ER stress remain unknown. Here we report that the gametophyte of P. patens displays severe growth inhibition and developmental deficiency under tunicamycin (Tm, an elicitor of ER stress)-induced ER stress conditions. PpAOX and selected ER stress response-like genes in P. patens were induced under Tm treatment. PpAOX knockout (PpAOX KO) plants exhibited decreased resistance to Tm-induced ER stress, whereas PpAOX-overexpressing lines (PpAOX OX) plants were more tolerant to Tm-induced ER stress. Data showed that PpAOX contributes to redox homeostasis under Tm treatment. In addition, we observed that PpAOX completely restores the Tm-sensitive phenotype of Arabidopsis AOX1a mutant (Ataox1a). Taken together, our work reveals a functional link between PpAOX and ER stress tolerance regulation in P. patens.

Black plastic film combined with straw mulching delays senescence and increases summer maize yield in northwest China

Ridge-furrow farming systems with transparent plastic film have been used extensively for maize (Zea mays L.) production in semi-arid areas. However, with climate warming, problems may arise associated with premature senescence and reduced production. This study examined whether dual mulching of ridges with black plastic film and furrows with straw can delay senescence and increase summer maize yield. Summer maize cultivation experiments were conducted in 2014–2015 by ridge-furrow planting with black (BRM) or transparent (TRM) plastic film mulch over the ridge and wheat straw mulch over the furrow, flat planting with black (BM) or transparent plastic film mulch (TM), and flat planting without mulch (control treatment, CK). The objective of this experiment was to determine the effects of the different mulching treatments on soil water and temperature, evapotranspiration (ET), leaf, photosynthesis rate, yield, and water use efficiency of summer maize. The BRM and TRM treatments were superior to the BM and TM treatments in terms of soil water storage in the 0–200 cm soil profile. The BRM reduced the diurnal variation in the soil temperature and lowered the soil temperature in the root zone. The BRM treatment extended the maize reproductive growth period by 5 days compared with TM, and 2–3 days longer than for the other mulched and control treatments. The BRM also increased the leaf area and dry matter accumulation per plant while significantly increasing the leaf photosynthesis rate. Moreover, BRM increased the 100–seed dry weight and significantly improved yield and water-use efficiency. The 2–year average yield of the BRM treatment was 6.7, 13.4, 18.4, and 72.9 % greater than yield of the TRM, BM, TM, and CK treatments, respectively. The corresponding water-use efficiency improved by 11.2, 13.1, 20.3, and 72.1 %, respectively. Applying the BRM cultivation practice will effectively delay senescence and increase summer maize yield in the agricultural production area of the study region.

Characterization of IRE1α in Neuro2a cells by pharmacological and CRISPR/Cas9 approaches.

IRE1 is the most conserved endoplasmic reticulum (ER)-resident stress sensor. Its activation not only splices XBP1 but also participates in a variety of cell signaling. We elucidated the role of IRE1α in Neuro2a cells by establishing IRE1α-deficient cells and applying four IRE1 inhibitors. IRE1α deficiency prevented almost all spliced XBP1 (sXBP1) protein expression by treatment with thapsigargin (Tg) and tunicamycin (Tm); these phenomena paralleled the values measured by our two Nanoluciferase-based IRE1 assays. However, cell viability and protein expression of other ER stress-responsive factors in the IRE1α-deficient cells were comparable to those in the parental wild-type cells with or without Tm treatment. Next, we elucidated the IRE1 inhibitory actions and cytotoxicity of four compounds: STF083010, KIRA6, 4μ8C, and toyocamycin. KIRA6 attenuated IRE1 activity in a dose-dependent manner, but it showed severe cytotoxicity even in the IRE1α-deficient cells at a low concentration. The IRE1α-deficient cells were slightly resistant to KIRA6 at 0.1μM in both the presence and absence of ER stress; however, resistance was not observed at 0.02μM. Treatment with only KIRA6 at 0.1μM for 12h remarkably induced LC3 II, an autophagic marker, in both parental and IRE1α-deficient cells. Co-treatment with KIRA6 and Tm induced LC3 II, cleaved caspase-9, and cleaved caspase-3; however, IRE1α-deficiency did not abolish the expression of these two cleaved caspases. On the other hand, KIRA6 prohibited Tm-induced ATF4 induction in an IRE1-independent manner; however, co-treatment with KIRA6 and Tm also induced LC3 II and two cleaved caspases in the ATF4-deficient Neuro2a cells. Thus, we demonstrate that IRE1α deficiency has little impact on cell viability and expression of ER stress-responsive factors in Neuro2a cells, and the pharmacological actions of KIRA6 include IRE1-independent ways.

Generation of a MOR-CreER knock-in mouse line to study cells and neural circuits involved in mu opioid receptor signaling.

Mu opioid receptor (MOR) is involved in various brain functions, such as pain modulation, reward processing, and addictive behaviors, and mediates the main pharmacologic effects of morphine and other opioid compounds. To gain genetic access to MOR-expressing cells, and to study physiological and pathological roles of MOR signaling, we generated a MOR-CreER knock-in mouse line, in which the stop codon of the Oprm1 gene was replaced by a DNA fragment encoding a T2A peptide and tamoxifen (Tm)-inducible Cre recombinase. We show that the MOR-CreER allele undergoes Tm-dependent recombination in a discrete subtype of neurons that express MOR in the adult nervous system, including the olfactory bulb, cerebral cortex, striosome compartments in the striatum, hippocampus, amygdala, thalamus, hypothalamus, interpeduncular nucleus, superior and inferior colliculi, periaqueductal gray, parabrachial nuclei, cochlear nucleus, raphe nuclei, pontine and medullary reticular formation, ambiguus nucleus, solitary nucleus, spinal cord, and dorsal root ganglia. The MOR-CreER mouse line combined with a Cre-dependent adeno-associated virus vector enables robust gene manipulation in the MOR-enriched striosomes. Furthermore, Tm treatment during prenatal development effectively induces Cre-mediated recombination. Thus, the MOR-CreER mouse is a powerful tool to study MOR-expressing cells with conditional gene manipulation in developing and mature neural tissues.

Unfolded Protein Response Differentially Regulates TLR4-Induced Cytokine Expression in Distinct Macrophage Populations.

Cellular stress responses are often engaged at sites of inflammation and can alter macrophage cytokine production. We now report that macrophages in distinct states of differentiation or in different temporal stages of inflammatory response exhibit differential sensitivity to cell stress mediated alterations in M1-like polarized inflammatory cytokine production. Tunicamycin (Tm) treatment of bone marrow derived macrophages (BMDM) cultured with M-CSF cultured bone marrow derived macrophages (M-BMDM) had markedly amplified M1-like responses to LPS, exhibiting higher levels of IL12p40 and IL12p35 mRNAs while BMDM cultured with GM-CSF, which normally express high IL12 subunit production in response to LPS, were relatively unaltered. Anti-inflammatory IL10 mRNA production in LPS-stimulated M-BMDM was greatly reduced by cell stress. These changes in cytokine mRNA levels resulted from altered rates of transcription and mRNA decay. Stress also altered cytokine protein production. Resident liver macrophages isolated from mice treated with Tm showed elevated levels of IL12 subunit mRNA production following LPS stimulation. Furthermore, macrophages infiltrating the liver during the early phase of acetaminophen injury (24 h) had little stress-mediated change in cytokine mRNA production while cells isolated in the later phase (48–72 h) exhibited higher sensitivity for stress elevated cytokine production. Hence cultured macrophages developed using different growth/differentiation factors and macrophages from different temporal stages of injury in vivo show markedly different sensitivity to cell stress for altered inflammatory cytokine production. These findings suggest that cellular stress can be an important modulator of the magnitude and character of myeloid inflammatory activity.

Transcriptional Regulation of Selenoprotein F by Heat Shock Factor 1 during Selenium Supplementation and Stress Response.

Changes of Selenoprotein F (SELENOF) protein levels have been reported during selenium supplementation, stressful, and pathological conditions. However, the mechanisms of how these external factors regulate SELENOF gene expression are largely unknown. In this study, HEK293T cells were chosen as an in vitro model. The 5′-flanking regions of SELENOF were analyzed for promoter features. Dual-Glo Luciferase assays were used to detect promoter activities. Putative binding sites of Heat Shock Factor 1 (HSF1) were predicted in silico and the associations were further proved by chromatin immunoprecipitation (ChIP) assay. Selenate and tunicamycin (Tm) treatment were used to induce SELENOF up-regulation. The fold changes in SELENOF expression and other relative proteins were analyzed by Q-PCR and western blot. Our results showed that selenate and Tm treatment up-regulated SELENOF at mRNA and protein levels. SELENOF 5′-flanking regions from −818 to −248 were identified as core positive regulatory element regions. Four putative HSF1 binding sites were predicted in regions from −1430 to −248, and six out of seven primers detected positive results in ChIP assay. HSF1 over-expression and heat shock activation increased the promoter activities, and mRNA and protein levels of SELENOF. Over-expression and knockdown of HSF1 showed transcriptional regulation effects on SELENOF during selenate and Tm treatment. In conclusion, HSF1 was discovered as one of the transcription factors that were associated with SELENOF 5′-flanking regions and mediated the up-regulation of SELENOF during selenate and Tm treatment. Our work has provided experimental data for the molecular mechanism of SELENOF gene regulation, as well as uncovered the involvement of HSF1 in selenotranscriptomic for the first time.

Medicago falcata MfSTMIR, an E3 ligase of endoplasmic reticulum-associated degradation, is involved in salt stress response.

SummaryRecent studies on E3 of endoplasmic reticulum (ER)‐associated degradation (ERAD) in plants have revealed homologs in yeast and animals. However, it remains unknown whether the plant ERAD system contains a plant‐specific E3 ligase. Here, we report that MfSTMIR, which encodes an ER‐membrane‐localized RING E3 ligase that is highly conserved in leguminous plants, plays essential roles in the response of ER and salt stress in Medicago. MfSTMIR expression was induced by salt and tunicamycin (Tm). mtstmir loss‐of‐function mutants displayed impaired induction of the ER stress‐responsive genes BiP1/2 and BiP3 under Tm treatment and sensitivity to salt stress. MfSTMIR promoted the degradation of a known ERAD substrate, CPY*. MfSTMIR interacted with the ERAD‐associated ubiquitin‐conjugating enzyme MtUBC32 and Sec61‐translocon subunit MtSec61γ. MfSTMIR did not affect MtSec61γ protein stability. Our results suggest that the plant‐specific E3 ligase MfSTMIR participates in the ERAD pathway by interacting with MtUBC32 and MtSec61γ to relieve ER stress during salt stress.

Deficiency of AtGFAT1 activity impairs growth, pollen germination and tolerance to tunicamycin in Arabidopsis.

The hexosamine biosynthetic pathway (HBP) plays essential roles in growth and development in plants. However, insight into the biological function of glutamine:fructose-6-phosphate amidotransferase 1 (GFAT1), mediating the first regulatory step of the HBP, remains unclear in plants. Here, we report the molecular characterization of Arabidopsis AtGFAT1 gene. AtGFAT1 was highly expressed in mature pollen grains, but its expression was not detectable in the rest of the organs. Pollen grains bearing the gfat1-2 knockout allele displayed defects in a polar deposition of pectin and callose in the pollen cell wall, leading to no genetic transmission of the gfat1-2 allele through the male gametophyte. AtGFAT1 overexpression increased glucosamine (GlcN) content and enhanced resistance to tunicamycin (Tm) treatment, while RNAi-mediated suppression reduced GlcN content and resistance to Tm treatment. However, the decrease in Tm resistance by RNAi suppression of AtGFAT1 was recovered by a GlcN supplement. The exogenous GlcN supplement also rescued gfat1-2/gaft1-2 mutant plants, which were otherwise not viable. The gfat1-2/gfat1-2 plants stopped growing at the germination stage on GlcN-free medium, but GlcN supplement allowed wild-type growth of gfat1-2/gfat1-2 plants. In addition, reactive oxygen species production, cell death and a decrease in protein N-glycosylation were observed in gfat1-2/gaft1-2 mutant plants grown on GlcN-free medium, whereas these aberrant defects were not detectable on GlcN-sufficient medium. Taken together, these results show that the reduction of protein N-glycosylation was at least partially responsible for many aberrant phenotypes in growth and development as well as the response to Tm treatment caused by AtGFAT1 deficiency in Arabidopsis.

Psychobiotics, as Promising Functional Food to Patients with Psychological Disorders: A Review on Mood Disorders, Sleep, and Cognition

Objective: Manipulating the intestinal microbiota for the benefit of the mental health is a concept that has become widely acknowledged. Prebiotics, the nondigestible nutrients, can proliferate intrinsic beneficial gut bacteria, and so provide an alternative strategy for effectively altering the enteric ecosystem, and then brain function. This review summarizes findings from studies using prebiotics to improve mental health and psychological disorders. Methods: Pub Med, Google Scholar, Scopus, and Science Direct databases were searched by using “prebiotics”, “psychological disorders”, “mood disorders”, “depression”, “anxiety”, “stress”, “sleep”, and “cognition” for searching the studies aiming the application of prebiotics and the beneficial effects of them in mental health and psychological disorders’ control and/or treatment. Results: Prebiotic consumption improved psychological and biological measures of mental disorders such as depression, anxiety, and stress in individuals with mood disorder. Overall, the results show that, through modulating the gut microbiota composition, prebiotics can beneficially affect mental health, modulate psychological disorders, and improve cognitive function and sleep/wake cycle disruptions. Discussion: Prebiotics can improve mental health, mood, and psychological function. Habitual diets rich in dietary prebiotics would be linked to reduced risk of developing symptoms of psychological disorders; however, additional studies are necessary.

Effects of mycorrhizal fungi inoculation and soil amendment with hydrogel on leaf anatomy, growth and physiology performance of olive plantlets under two contrasting water regimes

Abiotic stresses present a real environmental problem in agriculture field. In our paper, we examine the significance of arbuscular mycorrhizal fungi (AMF) and soil amendment with water retaining superpolymers (hydrogel) on growth and physiology performance of olive plantlets. Our experiment was carried out in nursery conditions, to test the impact of hydrogel (TH) and mycorrhizal fungi (TM), used individually or combined (THM), and compare them with non inoculated plants (TC), to understand and reduce the water stress damage in olive plantlets (cv. Chemlali). We also evaluate interactions between hydrogel, mycorrhizal treatments and water regimes. Results of mycorrhization (M%) show that roots colonized by Rhizophagus irregularis of well-watered plants were about 40.87%. In combined treatment (THM), M% was about 32.14%. Compared to TC treatment, TM treatment enhances significantly the dry weights of the whole plant under the two water regimes. The TM treatment had the highest relative water content (66.50%) and Chl (a + b) (0.83 mg g− 1) in stressed conditions. We found also that under water stress, the maximal quantum efficiency of the photosystem II measurements in leaves were significantly improved by 50.70% in TH treatment compared to control. For phenolic contents, TH treatment decreased significantly total phenols by 50.10% compared to TC. Our study gives evidence that the use of AMF and the hydrogel separately or in combination may enhance the capacity to avoid drought damages of olive plantlets and improve olive performances.

Effects of stem cell transplantation on bone mineral density and vitamin D status in children with thalassemia major.

HSCT is a curative treatment in TM, but conditioning and immunosuppressive treatment may affect bone metabolism. In this retrospective study, we aimed to compare BMD, vitamin D status, and growth in children with TM who underwent HSCT to those in children with TD TM. Twenty-three children with TM who underwent HSCT (mean age 7.1years [1.03-14.7]) and 24 children with TD thalassemia (mean age 9.8years [1.6-14]) were recruited. Lumbar spine BMD of TD thalassemia patients was higher than those in patients who had HSCT at both baseline and second-year assessments (P=.009, P<.001, respectively). However, BMD Z scores or serum 25-OH vitamin D levels were not different in two groups. Being >10years of age was a significant risk factor for low BMD, height, and weight Z score for both groups. Patients who underwent HSCT with Pesaro risk class II or III had higher risk for low BMD compared to those risk class I patients (P=.044). In conclusion, children with TM who were >10years at HSCT are at risk for low BMD and growth retardation. HSCT had no effect on BMD deficit in children with TM.

An Intelligent System Prototype to support and sharing diagnoses of maligned tumours, based on personalized medicine philosophy

Circulatory systems diseases are one of the most important causes of death in Chilean population according to a report presented by the Chilean National Bureau of Statistics (INE). Undoubtedly, these sad numbers arise an opportunity to analyse ways to improve this situation. Personalized Medicine is a new approach used to adapt standard medical treatments to individual characteristics of patients. Currently, several kinds of personalized-medicine software applications are building using Artificial Intelligent techniques and supported by techniques as Cloud Computing and Big Data. This architecture provides complex and varied information access, such as clinical data, genome data, patients’ treatment or drugs information, among others. This document describes a proposal to produce a method for generating and sharing medical information, particularly of maligned tumors in Chile. The prototype will be developed within the framework of the personalized medicine.

The Effects of Melatonin on Transcriptional Profile of Unfolded Protein Response Genes Under Endoplasmic Reticulum Stress in Arabidopsis thaliana

When the load of secretory pathway is increased or folding capacity in the endoplasmic reticulum (ER) is insufficient, unfolded proteins might accumulate in ER lumen causing a phenomenon called ER stress. During ER stress, normal cell functions are suppressed and unfolded protein response (UPR) is induced. Studies in animal systems suggest that melatonin alleviates the detrimental effects of ER stress; however, there is no study in plants in this respect. Hence, in this study, we investigated the possible role of melatonin on alleviation of ER stress in model plant Arabidopsis thaliana. Tunicamycin (Tm) was used to specifically induce ER stress. Melatonin treatment (10 and 25 μM but not 1 μM) increased root growth under Tm treatment, but it did not reach control levels. ER stress induced the expressions of ER stress sensor/transducer genes, ER chaperones and folding helper genes, ER-associated degradation (ERAD) genes, and ER stress-associated apoptosis genes in roots and shoots (a total of 16 genes). Among them, the expressions of ER stress sensor/transducer bZIP17, bZIP28, IRE1A, IRE1B, ERAD-related SEL1, and apoptosis genes AGB1 were decreased back to control levels with 25 μM melatonin under ER stress in roots. Moreover, Tm + melatonin treatments decreased the expressions of these genes when compared to only Tm-treated plants. Downregulation of UPR components with increased concentrations of melatonin under Tm treatment demonstrated that melatonin alleviated the detrimental effects of ER stress.

Tanshinone IIA ameliorates apoptosis of cardiomyocytes induced by endoplasmic reticulum stress.

The fat-soluble diterpenoids tanshinone IIA (TSA) is the major active element of Danshen, which has widespread cardioprotective effect. However, the mechanism of its beneficial effect on cardiomyocytes has not been fully investigated. Here, we aim to demonstrate that TSA ameliorates apoptosis of cardiomyocytes activated by endoplasmic reticulum stress (ERS). Primary cultures of neonatal rat cardiomyocytes are used, in which ERS-mediated apoptosis is induced by tunicamycin (Tm). Apoptosis of cardiomyocytes are detected by Hoechst staining and caspase 3 activity analysis. Protein expression of ERS markers are detected by Western blot, and level of miroRNA-133 (miR-133) is detected by real-time polymerase chain reaction. Tm treatment significantly triggers the apoptosis and ERS of cardiomyocytes. TSA dramatically ameliorates apoptosis and ERS of cardiomyocytes induced by Tm. Interestingly, level of miR-133 is reduced by Tm treatment, which is reversed by TSA. The cardioprotective effect of TSA on apoptosis and ERS of cardiomyocytes is blocked by anti-miR-133. These results suggest that TSA protects cardiomyocytes through ameliorated ERS-mediated apoptosis, which may be resulted from upregulation of miR-133.