Miracle-dz/UniFAR-FTUs huggingface dataset:
Autophagy Contributes to Leaf Starch Degradation[C][W]

Leaf starch degradation was thought to be confined to the chloroplasts, but this study demonstrates that autophagy, a non-plastid-based process, also contributes to the nighttime degradation of starch in leaves. This finding provides new insight into the biological role of autophagy and leaf starch degradation. Transitory starch, a major photosynthetic product in the leaves of land plants, accumulates in chloroplasts during the day and is hydrolyzed to maltose and Glc at night to support respiration and metabolism. Previous studies in Arabidopsis thaliana indicated that the degradation of transitory starch only occurs in the chloroplasts. Here, we report that autophagy, a nonplastidial process, participates in leaf starch degradation. Excessive starch accumulation was observed in Nicotiana benthamiana seedlings treated with an autophagy inhibitor and in autophagy-related (ATG) gene-silenced N. benthamiana and in Arabidopsis atg mutants. Autophagic activity in the leaves responded to the dynamic starch contents during the night. Microscopy showed that a type of small starch granule-like structure (SSGL) was localized outside the chloroplast and was sequestered by autophagic bodies. Moreover, an increased number of SSGLs was observed during starch depletion, and disruption of autophagy reduced the number of vacuole-localized SSGLs. These data suggest that autophagy contributes to transitory starch degradation by sequestering SSGLs to the vacuole for their subsequent breakdown.
MAP18 Regulates the Direction of Pollen Tube Growth in Arabidopsis by Modulating F-Actin Organization[C][W][OA]

MICROTUBULE-ASSOCIATED PROTEIN18 (MAP18) is known to regulate cortical microtubule organization and polar cell growth in vegetative tissues. This work shows that MAP18 is involved in the regulation of pollen tube morphology and directional growth for accurate micropyle targeting and fertilization by influencing actin organization, rather than microtubule assembly. For fertilization to occur in plants, the pollen tube must be guided to enter the ovule via the micropyle. Previous reports have implicated actin filaments, actin binding proteins, and the tip-focused calcium gradient as key contributors to polar growth of pollen tubes; however, the regulation of directional pollen tube growth is largely unknown. We reported previously that Arabidopsis thaliana MICROTUBULE-ASSOCIATED PROTEIN18 (MAP18) contributes to directional cell growth and cortical microtubule organization. The preferential expression of MAP18 in pollen and in pollen tubes suggests that MAP18 also may function in pollen tube growth. In this study, we demonstrate that MAP18 functions in pollen tubes by influencing actin organization, rather than microtubule assembly. In vitro biochemical results indicate that MAP18 exhibits Ca2+-dependent filamentous (F)-actin-severing activity. Abnormal expression of MAP18 in map18 and MAP18 OX plants was associated with disorganization of the actin cytoskeleton in the tube apex, resulting in aberrant pollen tube growth patterns and morphologies, inaccurate micropyle targeting, and fewer fertilization events. Experiments with MAP18 mutants created by site-directed mutagenesis suggest that F-actin-severing activity is essential to the effects of MAP18 on pollen tube growth direction. Our study demonstrates that in Arabidopsis, MAP18 guides the direction of pollen tube growth by modulating actin filaments.
Morphine Biosynthesis in Opium Poppy Involves Two Cell Types: Sieve Elements and Laticifers[W][OPEN]

Morphine biosynthesis is shown to involve two specialized cell types in opium poppy. Most of the pathway occurs in sieve elements of the phloem, but the final three enzymes are predominant in adjacent laticifers, which are the site of morphine accumulation. The cellular localization of morphine metabolism will assist with the breeding and metabolic engineering of this important medicinal plant. Immunofluorescence labeling and shotgun proteomics were used to establish the cell type–specific localization of morphine biosynthesis in opium poppy (Papaver somniferum). Polyclonal antibodies for each of six enzymes involved in converting (R)-reticuline to morphine detected corresponding antigens in sieve elements of the phloem, as described previously for all upstream enzymes transforming (S)-norcoclaurine to (S)-reticuline. Validated shotgun proteomics performed on whole-stem and latex total protein extracts generated 2031 and 830 distinct protein families, respectively. Proteins corresponding to nine morphine biosynthetic enzymes were represented in the whole stem, whereas only four of the final five pathway enzymes were detected in the latex. Salutaridine synthase was detected in the whole stem, but not in the latex subproteome. The final three enzymes converting thebaine to morphine were among the most abundant active latex proteins despite a limited occurrence in laticifers suggested by immunofluorescence labeling. Multiple charge isoforms of two key O-demethylases in the latex were revealed by two-dimensional immunoblot analysis. Salutaridine biosynthesis appears to occur only in sieve elements, whereas conversion of thebaine to morphine is predominant in adjacent laticifers, which contain morphine-rich latex. Complementary use of immunofluorescence labeling and shotgun proteomics has substantially resolved the cellular localization of morphine biosynthesis in opium poppy.
Postoperative Alcohol Withdrawal Syndrome and Neuropsychological Disorder in Patients after Head and Neck Cancer Ablation Followed by Microsurgical Free Tissue Transfer

Abstract Purpose The use of microsurgical free flap reconstruction has resulted in improved patient outcomes and survival after head and neck cancer resection. Although postoperative care in an intensive care unit (ICU) for airway management and flap surveillance can increase the success rate, any accompanying neuropsychological problems can potentially affect the outcome. Materials and Methods From January 2006 to December 2008, we retrospectively reviewed 1,506 ICU patients who underwent head and neck cancer ablative surgery followed by microsurgical free tissue transfer. Twelve patients (Group A) had alcohol withdrawal syndrome (AWS), whereas 29 patients (Group B) had other neuropsychological problems. The clinical manifestations and outcomes of each group were investigated and statically analyzed. Results All the flaps survived. There was no significant difference in onset time, duration, ICU stay, hospital stay interval, overall complications, and flap circulation-related complications. However, the non–flap-related complication rates (10/12, 83.3%; 14/29, 48.3%; p = 0.038), including failure of extubation or ventilator weaning (7/12, 58.3%; 7/29, 24.1%; p = 0.036), were significantly higher in Group A. Conclusion Patients with postoperative AWS have a higher chance of developing non–flap-related complications, especially respiratory problems. The identification of a multidisciplinary approach to identify and manage these patients preoperatively and postoperatively is required.
The effects of prenatal exposure to a ‘junk food’ diet on offspring food preferences and fat deposition can be mitigated by improved nutrition during lactation

Exposure to a maternal junk food (JF) diet in utero and during the suckling period has been demonstrated to increase the preference for palatable food and increase the susceptibility to diet-induced obesity in adult offspring. We aimed to determine whether the effects of prenatal exposure to JF could be ameliorated by cross-fostering offspring onto dams consuming a standard rodent chow during the suckling period. We report here that when all offspring were given free access to the JF diet for 7 weeks from 10 weeks of age, male offspring of control (C) or JF dams that were cross-fostered at birth onto JF dams (C-JF, JF-JF), exhibited higher fat (C-C: 12.3 ± 0.34 g/kg/day; C-JF: 14.7 ± 1.04 g/kg/day; JF-C: 11.5 ± 0.41 g/kg/day; JF-JF: 14.0 ± 0.44 g/kg/day; P < 0.05) and overall energy intake (C-C: 930.1 ± 18.56 kJ/kg/day; C-JF: 1029.0 ± 82.9 kJ/kg/day; JF-C: 878.3 ± 19.5 kJ/kg/day; JF-JF: 1003.4 ± 25.97 kJ/kg/day; P < 0.05) than offspring exposed to the JF diet only before birth (JF-C) or not at all (C-C). Female offspring suckled by JF dams, despite no differences in food intake, had increased fat mass as percentage of body weight (C-C: 19.9 ± 1.33%; C-JF: 22.8 ± 1.57%; JF-C: 17.4 ± 1.03%; JF-JF: 22.0 ± 1.0%; P < 0.05) after 3 weeks on the JF diet. No difference in fat mass was observed in male offspring. These findings suggest that the effects of prenatal exposure to a JF diet on food preferences in females and susceptibility to diet-induced obesity in males can be prevented by improved nutrition during the suckling period.
Arabidopsis Chlorophyll Biosynthesis: An Essential Balance between the Methylerythritol Phosphate and Tetrapyrrole Pathways[C][W]

This work provides evidence into how inhibition of the methylerythritol phosphate pathway causes plant death. Chlorophyll is composed of a chlorin ring, generated by the tetrapyrrole pathway, and an isoprenoid tail, made from the methylerythritol phosphate pathway. The data herein demonstrate that stoichiometric balance between these pathways is essential for viability. Chlorophyll, essential for photosynthesis, is composed of a chlorin ring and a geranylgeranyl diphosphate (GGPP)–derived isoprenoid, which are generated by the tetrapyrrole and methylerythritol phosphate (MEP) biosynthesis pathways, respectively. Although a functional MEP pathway is essential for plant viability, the underlying basis of the requirement has been unclear. We hypothesized that MEP pathway inhibition is lethal because a reduction in GGPP availability results in a stoichiometric imbalance in tetrapyrrolic chlorophyll precursors, which can cause deadly photooxidative stress. Consistent with this hypothesis, lethality of MEP pathway inhibition in Arabidopsis thaliana by fosmidomycin (FSM) is light dependent, and toxicity of MEP pathway inhibition is reduced by genetic and chemical impairment of the tetrapyrrole pathway. In addition, FSM treatment causes a transient accumulation of chlorophyllide and transcripts associated with singlet oxygen-induced stress. Furthermore, exogenous provision of the phytol molecule reduces FSM toxicity when the phytol can be modified for chlorophyll incorporation. These data provide an explanation for FSM toxicity and thereby provide enhanced understanding of the mechanisms of FSM resistance. This insight into MEP pathway inhibition consequences underlines the risk plants undertake to synthesize chlorophyll and suggests the existence of regulation, possibly involving chloroplast-to-nucleus retrograde signaling, that may monitor and maintain balance of chlorophyll precursor synthesis.
No associations of prenatal maternal psychosocial stress with fasting glucose metabolism in offspring at 5–6 years of age

Highly prevalent maternal psychosocial complaints are accompanied by increases in glucocorticoid stress hormones, which may predispose the offspring for type 2 diabetes and cardiovascular disease later in adulthood. The aim of the current research is to study whether prenatal maternal psychosocial stress is associated with parameters of blood glucose metabolism in their children aged 5–6 years. The study design was a prospective birth cohort (the Amsterdam Born Children and their Development study, the Netherlands). Depressive symptoms, pregnancy-related anxiety, parenting daily hassles and job strain were recorded by questionnaire (gestational week 16). A cumulative score was also calculated. Possible sex differences in the associations were considered. The subjects were 1952 mother–child pairs. Outcome measures were fasting glucose (n=1952), C-peptide and insulin resistance (HOMA2-IR) (n=1478) in the children at the age of 5–6 years. The stress scales, single and cumulative, were not associated with glucose/C-peptide/insulin resistance (all P>0.05). We did not find evidence for sex differences. In conclusion, we did not find evidence for an association between psychosocial stress during early pregnancy and parameters of glucose metabolism in offspring at the age of 5–6 years. Differences emerging later in life or in response to a metabolic challenge should not be ruled out.
Improving allogeneic islet transplantation by suppressing Th17 and enhancing Treg with histone deacetylase inhibitors

Islet transplantation is a new treatment for achieving insulin independence for patients with severe diabetes. However, major drawbacks of this treatment are the long graft survival, the necessity for immunosuppressive drugs, and the efficacy of transplantation. Donor‐specific transfusion (DST) has been shown to reduce rejection after organ transplantation, potentially through enhanced regulatory T‐cell (Treg) activity. However, recent findings have shown that activated Treg can be converted into Th17 cells. We focused on histone deacetylase inhibitors (HDACi) because it was reported that inhibition of HDAC activity prevented Treg differentiation into IL17‐producing cells. We therefore sought to enhance Treg while suppressing Th17 cells using DST with HDACi to prolong graft survival. To stimulate Treg by DST, we used donor splenocytes. In DST with HDACi group, Foxp3 mRNA expression and Treg population increased in the thymus and spleen, whereas Th17 population decreased. qPCR analysis of lymphocyte mRNA indicated that Foxp3, IL‐10, and TGF‐b expression increased. However, interleukin 17a, Stat3 (Th17), and IFN‐g expression decreased in DST + HDACi group, relative to DST alone. Moreover, DST treated with HDACi prolonged graft survival relative to controls in mice islet transplantation. DST with HDACi may therefore have utility in islet transplantation.
The Arabidopsis Ethylene/Jasmonic Acid-NRT Signaling Module Coordinates Nitrate Reallocation and the Trade-Off between Growth and Environmental Adaptation[W][OPEN]

Stress-induced nitrate allocation to roots serves as a universal mechanism in response to diverse stresses, although how it perceives the environment remains unknown. This study demonstrates that the ET/JA-NRT1.5/NRT1.8 signaling module functions in this perception to fine-tune nitrate allocation and plant adaptation to the environment. Stresses decouple nitrate assimilation and photosynthesis through stress-initiated nitrate allocation to roots (SINAR), which is mediated by the nitrate transporters NRT1.8 and NRT1.5 and functions to promote stress tolerance. However, how SINAR communicates with the environment remains unknown. Here, we present biochemical and genetic evidence demonstrating that in Arabidopsis thaliana, ethylene (ET) and jasmonic acid (JA) affect the crosstalk between SINAR and the environment. Electrophoretic mobility shift assays and chromatin immunoprecipitation assays showed that ethylene response factors (ERFs), including OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF59, bind to the GCC boxes in the NRT1.8 promoter region, while ETHYLENE INSENSITIVE3 (EIN3) binds to the EIN3 binding site motifs in the NRT1.5 promoter. Genetic assays showed that cadmium and sodium stresses initiated ET/JA signaling, which converged at EIN3/EIN3-Like1 (EIL1) to modulate ERF expression and hence to upregulate NRT1.8. By contrast, ET and JA signaling mediated the downregulation of NRT1.5 via EIN3/EIL1 and other, unknown component(s). SINAR enhanced stress tolerance and decreased plant growth under nonstressed conditions through the ET/JA-NRT1.5/NRT1.8 signaling module. Interestingly, when nitrate reductase was impaired, SINAR failed to affect either stress tolerance or plant growth. These data suggest that SINAR responds to environmental conditions through the ET/JA-NRT signaling module, which further modulates stress tolerance and plant growth in a nitrate reductase-dependent manner.
The Ubiquitin Receptors DA1, DAR1, and DAR2 Redundantly Regulate Endoreduplication by Modulating the Stability of TCP14/15 in Arabidopsis

A genetic and molecular mechanism involving the ubiquitin receptors DA1, DAR1, and DAR2 and the transcription factors TCP14 and TCP15 links cell and organ growth with the regulation of endoreduplication. Organ growth involves the coordination of cell proliferation and cell growth with differentiation. Endoreduplication is correlated with the onset of cell differentiation and with cell and organ size, but little is known about the molecular mechanisms linking cell and organ growth with endoreduplication. We have previously demonstrated that the ubiquitin receptor DA1 influences organ growth by restricting cell proliferation. Here, we show that DA1 and its close family members DAR1 and DAR2 are redundantly required for endoreduplication during leaf development. DA1, DAR1, and DAR2 physically interact with the transcription factors TCP14 and TCP15, which repress endoreduplication by directly regulating the expression of cell-cycle genes. We also show that DA1, DAR1, and DAR2 modulate the stability of TCP14 and TCP15 proteins in Arabidopsis thaliana. Genetic analyses demonstrate that DA1, DAR1, and DAR2 function in a common pathway with TCP14/15 to regulate endoreduplication. Thus, our findings define an important genetic and molecular mechanism involving the ubiquitin receptors DA1, DAR1, and DAR2 and the transcription factors TCP14 and TCP15 that links endoreduplication with cell and organ growth.
Deficiency of the Arabidopsis Helicase RTEL1 Triggers a SOG1-Dependent Replication Checkpoint in Response to DNA Cross-Links

Deficiency of the Arabidopsis RTEL1 DNA helicase activates a replication-dependent cell cycle arrest that depends on the SOG1 checkpoint regulator. To maintain genome integrity, DNA replication is executed and regulated by a complex molecular network of numerous proteins, including helicases and cell cycle checkpoint regulators. Through a systematic screening for putative replication mutants, we identified an Arabidopsis thaliana homolog of human Regulator of Telomere Length 1 (RTEL1), which functions in DNA replication, DNA repair, and recombination. RTEL1 deficiency retards plant growth, a phenotype including a prolonged S-phase duration and decreased cell proliferation. Genetic analysis revealed that rtel1 mutant plants show activated cell cycle checkpoints, specific sensitivity to DNA cross-linking agents, and increased homologous recombination, but a lack of progressive shortening of telomeres, indicating that RTEL1 functions have only been partially conserved between mammals and plants. Surprisingly, RTEL1 deficiency induces tolerance to the deoxynucleotide-depleting drug hydroxyurea, which could be mimicked by DNA cross-linking agents. This resistance does not rely on the essential replication checkpoint regulator WEE1 but could be blocked by a mutation in the SOG1 transcription factor. Taken together, our data indicate that RTEL1 is required for DNA replication and that its deficiency activates a SOG1-dependent replication checkpoint.
Arabidopsis Chloroplast Mini-Ribonuclease III Participates in rRNA Maturation and Intron Recycling

Using RNA-Seq and end-mapping, we showed that two Arabidopsis Mini-III proteins, which are putative double-stranded plastid endonucleases, process rRNA, introns, and a plastid noncoding RNA. RNase III proteins recognize double-stranded RNA structures and catalyze endoribonucleolytic cleavages that often regulate gene expression. Here, we characterize the functions of RNC3 and RNC4, two Arabidopsis thaliana chloroplast Mini-RNase III-like enzymes sharing 75% amino acid sequence identity. Whereas rnc3 and rnc4 null mutants have no visible phenotype, rnc3/rnc4 (rnc3/4) double mutants are slightly smaller and chlorotic compared with the wild type. In Bacillus subtilis, the RNase Mini-III is integral to 23S rRNA maturation. In Arabidopsis, we observed imprecise maturation of 23S rRNA in the rnc3/4 double mutant, suggesting that exoribonucleases generated staggered ends in the absence of specific Mini-III-catalyzed cleavages. A similar phenotype was found at the 3′ end of the 16S rRNA, and the primary 4.5S rRNA transcript contained 3′ extensions, suggesting that Mini-III catalyzes several processing events of the polycistronic rRNA precursor. The rnc3/4 mutant showed overaccumulation of a noncoding RNA complementary to the 4.5S-5S rRNA intergenic region, and its presence correlated with that of the extended 4.5S rRNA precursor. Finally, we found rnc3/4-specific intron degradation intermediates that are probable substrates for Mini-III and show that B. subtilis Mini-III is also involved in intron regulation. Overall, this study extends our knowledge of the key role of Mini-III in intron and noncoding RNA regulation and provides important insight into plastid rRNA maturation.
Women and Men With Mental Illnesses

Women with mental illnesses have reported that one painful aspect of living with their diagnosis is the loss of personal identity when they are labeled “mentally ill. ” Although a number of studies have focused on services to address issues of relevance to women, few have explored the different ways in which women and men view services within the mental health system. Using a focus-group approach, the authors conducted a study that examined the service needs and views of women and men with mental illnesses. This article reports on this study and identifies the participants' common and unique needs and views and discusses the implications for research and practice.
“Ward talk”: Nurses’ interaction with people with and without aphasia in the very early period poststroke

Background: Nursing staff are the most frequent communication partners, after family members, for people in hospital poststroke, and they play an essential role in the multidisciplinary team. Recent research has found that patients are more cognitively and socially active when wards provide an “enriched environment” as compared to standard care. Therefore, language enrichment on acute wards is now being considered as a possible way to discourage “learned nonuse” of language in people with aphasia. Aims: This study involved an exploration of the nature of nurses’ interactions with three patients on an acute stroke ward, two with aphasia and one without, in order to understand the nature of the communicative environment in more detail. Methods & Procedures: Following all necessary ethics approval processes and consents, continuous video recordings of 7.5 hr each were run for three male patients (two aged 68 years and one aged 48 years) on the acute stroke ward within a teaching hospital in an Australian city. Two had left hemisphere strokes resulting in a Wernicke’s aphasia (WAB AQ = 47) and a global aphasia (WAB AQ = 9.2) and one had a right hemisphere stroke and no aphasia. All instances of interaction with nurses over that period were transcribed orthographically and analysed at a discourse level using a speech function analysis based on a systemic functional linguistic framework. Outcomes & Results: Overall, nurses made most of the opening moves, used closed questions, controlled the conversational floor, and did not generally interact about issues other than physical care. The patients with aphasia had restricted opportunity to use continuing and developing moves and most commonly fell into a pattern of closed question and simple response. There was little evidence of communicative repair by nurses and few supported conversation strategies employed. Patterns of moves identified through the speech function analysis revealed the disempowered position of the patients with aphasia compared to the patient without who used a wider range of speech functions that demonstrated more assertive interactions. Conclusions: This study suggests that nurses, in their role as key communication partners on the acute stroke ward, could help counter the development of learned nonuse by integrating more effective communication strategies into their daily care routines. Further conversation partner training could relieve communication breakdown and frustration and increase opportunities for satisfying conversational exchanges in this setting.