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AIMS: We investigated the characteristics of infection and the utility of inflammatory markers in diabetic ketoacidosis (DKA) and hyperosmolar hyperglycemic syndrome (HHS). METHODS: A multicenter, retrospective observational study in 21 acute-care hospitals was conducted in Japan. This study included adult hospitalized patients with DKA and HHS. We analyzed the diagnostic accuracy of markers including C-reactive protein (CRP) and procalcitonin (PCT) for bacteremia. Multiple regression models were created for estimating bacteremia risk factors. RESULTS: A total of 771 patients, including 545 patients with DKA and 226 patients with HHS, were analyzed. The mean age was 58.2 (SD, 19.3) years. Of these, 70 tested positive for blood culture. The mortality rates of those with and without bacteremia were 14 % and 3.3 % (P-value < 0.001). The area under the curve (AUC) of CRP and PCT for diagnosis of bacteremia was 0.85 (95 %CI, 0.81-0.89) and 0.76 (95 %CI, 0.60-0.92), respectively. Logistic regression models identified older age, altered level of consciousness, hypotension, and higher CRP as risk factors for bacteremia. CONCLUSIONS: The mortality rate was higher in patients with bacteremia than patients without it. CRP, rather than PCT, may be valid for diagnosing bacteremia in hyperglycemic emergencies. TRIAL REGISTRATION: This study is registered in the UMIN clinical trial registration system (UMIN000025393, Registered December 23, 2016).
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Bacteriemia , Proteína C-Reativa , Cetoacidose Diabética , Coma Hiperglicêmico Hiperosmolar não Cetótico , Humanos , Estudos Retrospectivos , Masculino , Feminino , Pessoa de Meia-Idade , Cetoacidose Diabética/diagnóstico , Cetoacidose Diabética/sangue , Cetoacidose Diabética/epidemiologia , Coma Hiperglicêmico Hiperosmolar não Cetótico/diagnóstico , Coma Hiperglicêmico Hiperosmolar não Cetótico/sangue , Coma Hiperglicêmico Hiperosmolar não Cetótico/complicações , Idoso , Adulto , Bacteriemia/diagnóstico , Bacteriemia/mortalidade , Bacteriemia/epidemiologia , Proteína C-Reativa/análise , Proteína C-Reativa/metabolismo , Japão/epidemiologia , Fatores de Risco , Pró-Calcitonina/sangue , Biomarcadores/sangueRESUMO
Hyperglycemic emergencies frequently lead to acute kidney injury (AKI) and require treatment with large amount of intravenous fluids. However, the effects of chloride loading on this population have not yet been investigated. We conducted a multicenter, retrospective, cohort study in 21 acute-care hospitals in Japan. The study included hospitalized adult patients with diabetic ketoacidosis (DKA) and hyperosmolar hyperglycemic syndrome (HHS) who had AKI upon arrival. The patients were classified into high and low chloride groups based on the amount of chloride administered within the first 48 h of their arrival. The primary outcome was recovery from AKI; secondary outcome was major adverse kidney events within 30 days (MAKE30), including mortality and prolonged renal failure. A total of 390 patients with AKI, including 268 (69%) with DKA and 122 (31%) with HHS, were included in the study. Using the criteria of Kidney Disease Improving Global Outcomes, the severity of AKI in the patients was Stage 1 (n = 159, 41%), Stage 2 (n = 121, 31%), and Stage 3 (n = 110, 28%). The analysis showed no significant difference between the two groups in recovery from AKI (adjusted hazard ratio, 0.96; 95% CI 0.72-1.28; P = 0.78) and in MAKE30 (adjusted odds ratio, 0.91; 95% CI 0.45-1.76; P = 0.80). Chloride loading with fluid administration had no significant impact on recovery from AKI in patients with hyperglycemic emergencies.Trial Registration This study was registered in the UMIN clinical trial registration system (UMIN000025393, registered December 23, 2016).
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Injúria Renal Aguda , Cetoacidose Diabética , Humanos , Estudos Retrospectivos , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/terapia , Injúria Renal Aguda/fisiopatologia , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Japão/epidemiologia , Cetoacidose Diabética/complicações , Cloretos/sangue , Cloretos/análise , Estudos de Coortes , Adulto , Hiperglicemia/complicações , Coma Hiperglicêmico Hiperosmolar não Cetótico/complicações , Hidratação/métodos , EmergênciasRESUMO
In Catharanthus roseus, monoterpenoid indole alkaloids (MIAs) are produced through the cooperation of four cell types, with final products accumulating in specialized cells known as idioblasts and laticifers. To explore the relationship between cellular differentiation and cell type-specific MIA metabolism, we analyzed the expression of MIA biosynthesis in germinating seeds. Embryos from immature and mature seeds were observed via stereomicroscopy, fluorescence microscopy, and electron microscopy. Time-series MIA and iridoid quantification, along with transcriptome analysis, were conducted to determine the initiation of MIA biosynthesis. In addition, the localization of MIAs was examined using alkaloid staining and imaging mass spectrometry (IMS). Laticifers were present in embryos before seed maturation. MIA biosynthesis commenced 12 h after germination. MIAs accumulated in laticifers of embryos following seed germination, and MIA metabolism is induced after germination in a tissue-specific manner. These findings suggest that cellular morphological differentiation precedes metabolic differentiation. Considering the well-known toxicity and defense role of MIAs in matured plants, MIAs may be an important defense strategy already in the delicate developmental phase of seed germination, and biosynthesis and accumulation of MIAs may require the tissue and cellular differentiation.
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Catharanthus , Alcaloides de Triptamina e Secologanina , Monoterpenos/metabolismo , Catharanthus/metabolismo , Germinação , Sementes/metabolismo , Alcaloides de Triptamina e Secologanina/metabolismo , Diferenciação Celular , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
Due to their size and locomotion ability, cockroaches are favorable as hybrid robot platforms in search and rescue (SAR) missions. However, cockroaches most likely approach the corner area and stay for an uncertain time. This natural behavior will hinder the utilization of cyborg cockroaches in SAR missions under rubble, unstructured, and unknown areas. Therefore, we proposed onboard automatic obstacle avoidance and human detection that can run on the wireless backpack stimulator without an external motion capture system. A low-power and small-size Time of Flight (ToF) sensor was selected as a distance measurement sensor, while a low-resolution thermopile array sensor was applied for human presence detection. The implemented feedback control based on IMU and ToF sensors has successfully navigated the cyborg cockroach to avoid obstacles and escape from the sharp corners in the laboratory unstructured area without stopping or being trapped. It could also recognize the human presence when the human was in front of it in real-time. Due to its performance, the random forest classifier was implemented as an embedded human detection system. It could achieve the highest accuracy at a distance of around 25 cm (92.5%) and the lowest accuracy at about 100 cm (70%).
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INTRODUCTION: Diabetic ketoacidosis (DKA) and hyperglycemic hyperosmolar syndrome (HHS) are life-threatening complications of diabetes mellitus. Their clinical profiles have not been fully investigated. METHODS: A multicenter retrospective cohort study was conducted in 21 acute care hospitals in Japan. Patients included were adults aged 18 or older who had been hospitalized from January 1, 2012, to December 31, 2016 due to DKA or HHS. The data were extracted from patient medical records. A four-group comparison (mild DKA, moderate DKA, severe DKA, and HHS) was performed to evaluate outcomes. RESULTS: A total of 771 patients including 545 patients with DKA and 226 patients with HHS were identified during the study period. The major precipitating factors of disease episodes were poor medication compliance, infectious diseases, and excessive drinking of sugar-sweetened beverages. The median hospital stay was 16 days [IQR 10-26 days]. The intensive care unit (ICU) admission rate was 44.4% (mean) and the rate at each hospital ranged from 0 to 100%. The in-hospital mortality rate was 2.8% in patients with DKA and 7.1% in the HHS group. No significant difference in mortality was seen among the three DKA groups. CONCLUSIONS: The mortality rate of patients with DKA in Japan is similar to other studies, while that of HHS was lower. The ICU admission rate varied among institutions. There was no significant association between the severity of DKA and mortality in the study population. TRIAL REGISTRATION: This study is registered in the UMIN clinical Trial Registration System (UMIN000025393, Registered 23th December 2016).
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Diabetes Mellitus , Cetoacidose Diabética , Coma Hiperglicêmico Hiperosmolar não Cetótico , Adulto , Humanos , Cetoacidose Diabética/etiologia , Cetoacidose Diabética/complicações , Coma Hiperglicêmico Hiperosmolar não Cetótico/complicações , Coma Hiperglicêmico Hiperosmolar não Cetótico/epidemiologia , Estudos Retrospectivos , Japão/epidemiologia , HospitaisRESUMO
Purpose To evaluate the feasibility of liver MR fingerprinting (MRF) for quantitative characterization and diagnosis of focal liver lesions. Materials and Methods This single-site, prospective study included 89 participants (mean age, 62 years ± 15 [SD]; 45 women, 44 men) with various focal liver lesions who underwent MRI between October 2021 and August 2022. The participants underwent routine clinical MRI, non-contrast-enhanced liver MRF, and reference quantitative MRI with a 1.5-T MRI scanner. The bias and repeatability of the MRF measurements were assessed using linear regression, Bland-Altman plots, and coefficients of variation. The diagnostic capability of MRF-derived T1, T2, T2*, proton density fat fraction (PDFF), and a combination of these metrics to distinguish benign from malignant lesions was analyzed according to the area under the receiver operating characteristic curve (AUC). Results Liver MRF measurements showed moderate to high agreement with reference measurements (intraclass correlation = 0.94, 0.77, 0.45, and 0.61 for T1, T2, T2*, and PDFF, respectively), with underestimation of T2 values (mean bias in lesion = -0.5%, -29%, 5.8%, and -8.2% for T1, T2, T2*, and PDFF, respectively). The median coefficients of variation for repeatability of T1, T2, and T2* values were 2.5% (IQR, 3.6%), 3.1% (IQR, 5.6%), and 6.6% (IQR, 13.9%), respectively. After considering multicollinearity, a combination of MRF measurements showed a high diagnostic performance in differentiating benign from malignant lesions (AUC = 0.92 [95% CI: 0.86, 0.98]). Conclusion Liver MRF enabled the quantitative characterization of various focal liver lesions in a single breath-hold acquisition. Keywords: MR Imaging, Abdomen/GI, Liver, Imaging Sequences, Technical Aspects, Tissue Characterization, Technology Assessment, Diagnosis, Liver Lesions, MR Fingerprinting, Quantitative Characterization Supplemental material is available for this article. © RSNA, 2023.
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Neoplasias Hepáticas , Imageamento por Ressonância Magnética , Masculino , Humanos , Feminino , Pessoa de Meia-Idade , Estudos Prospectivos , Imageamento por Ressonância Magnética/métodos , Abdome , Prótons , Neoplasias Hepáticas/diagnóstico por imagemRESUMO
Arabidopsis ASYMMETRIC LEAVES2 (AS2) plays a key role in the formation of flat symmetric leaves. AS2 represses the expression of the abaxial gene ETTIN/AUXIN RESPONSE FACTOR3 (ETT/ARF3). AS2 interacts in vitro with the CGCCGC sequence in ETT/ARF3 exon 1. In cells of leaf primordia, AS2 localizes at peripheral regions of the nucleolus as two AS2 bodies, which are partially overlapped with chromocenters that contain condensed 45S ribosomal DNA repeats. AS2 contains the AS2/LOB domain, which consists of three sequences conserved in the AS2/LOB family: the zinc finger (ZF) motif, the ICG sequence including the conserved glycine residue, and the LZL motif. AS2 and the genes NUCLEOLIN1 (NUC1), RNA HELICASE10 (RH10), and ROOT INITIATION DEFECTIVE2 (RID2) that encode nucleolar proteins coordinately act as repressors against the expression of ETT/ARF3. Here, we examined the formation and patterning of AS2 bodies made from as2 mutants with amino acid substitutions in the ZF motif and the ICG sequence in cells of cotyledons and leaf primordia. Our results showed that the amino acid residues next to the cysteine residues in the ZF motif were essential for both the formation of AS2 bodies and the interaction with ETT/ARF3 DNA. The conserved glycine residue in the ICG sequence was required for the formation of AS2 bodies, but not for the DNA interaction. We also examined the effects of nuc1, rh10, and rid2 mutations, which alter the metabolism of rRNA intermediates and the morphology of the nucleolus, and showed that more than two AS2 bodies were observed in the nucleolus and at its periphery. These results suggested that the patterning of AS2 bodies is tightly linked to the morphology and functions of the nucleolus and the development of flat symmetric leaves in plants.
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Advances in omics technologies now permit the generation of highly contiguous genome assemblies, detection of transcripts and metabolites at the level of single cells and high-resolution determination of gene regulatory features. Here, using a complementary, multi-omics approach, we interrogated the monoterpene indole alkaloid (MIA) biosynthetic pathway in Catharanthus roseus, a source of leading anticancer drugs. We identified clusters of genes involved in MIA biosynthesis on the eight C. roseus chromosomes and extensive gene duplication of MIA pathway genes. Clustering was not limited to the linear genome, and through chromatin interaction data, MIA pathway genes were present within the same topologically associated domain, permitting the identification of a secologanin transporter. Single-cell RNA-sequencing revealed sequential cell-type-specific partitioning of the leaf MIA biosynthetic pathway that, when coupled with a single-cell metabolomics approach, permitted the identification of a reductase that yields the bis-indole alkaloid anhydrovinblastine. We also revealed cell-type-specific expression in the root MIA pathway.
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Antineoplásicos , Catharanthus , Plantas Medicinais , Catharanthus/genética , Plantas Medicinais/metabolismo , Multiômica , Alcaloides Indólicos/metabolismo , Antineoplásicos/metabolismo , Monoterpenos/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
MAIN CONCLUSION: Using virus-induced gene silencing, we demonstrated that the enzymes GES, ISY, and MLPL are responsible for nepetalactone biosynthesis in Nepeta cataria. Nepetalactone is the main iridoid that is found in the Nepeta genus and is well-known for its psychoactive effect on house cats. Moreover, there is a burgeoning interest into the effect of nepetalactone on insects. Although the enzymes for nepetalactone biosynthesis have been biochemically assayed in vitro, validation of the role that these enzymes have in planta has not been demonstrated. Virus-induced gene silencing (VIGS) is a silencing method that relies on transient transformation and is an approach that has been particularly successful when applied to a variety of non-model plants. Here, we use a recently designed visual-marker dependent VIGS system to demonstrate that the nepetalactone biosynthetic enzymes GES, ISY, and MLPL impact nepetalactone biosynthesis in Nepeta cataria.
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Nepeta , Monoterpenos Ciclopentânicos , Iridoides , Nepeta/química , Nepeta/genética , Pironas/química , Pironas/farmacologiaRESUMO
To understand how the plant regulates metabolism, it is important to determine where metabolites localize in the tissues and cells. Single-cell level omics approaches in plants have shown remarkable development over the last several years, and this data has been instrumental in gene discovery efforts for enzymes and transporters involved in metabolism. For metabolomics, Imaging Mass Spectrometry (IMS) is a powerful tool to map the spatial distribution of molecules in the tissue. Here, we describe the methods which we used to reveal where secondary metabolites, primarily alkaloids, localize in Catharanthus roseus stem and leaf tissues.
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Alcaloides , Catharanthus , Alcaloides/análise , Catharanthus/metabolismo , Espectrometria de Massas , Metabolômica/métodos , Folhas de Planta/metabolismoRESUMO
Bioactive specialized (secondary) metabolites are indispensable for plant development or adjustment to their surrounding environment. In many plants, these specialized metabolites are accumulated in specifically differentiated cells. Catharanthus roseus is a well-known medicinal plant known for producing many kinds of monoterpenoid indole alkaloids (MIAs). C. roseus has two types of specifically differentiated cells accumulating MIAs, so-called idioblast cells and laticifer cells. In this study, we compared each of the cells as they changed during seedling growth, and found that the fluorescent metabolites accumulated in these cells were differentially regulated. Analysis of fluorescent compounds revealed that the fluorescence observed in these cells was emitted from the compound serpentine. Further, we found that the serpentine content of leaves increased as leaves grew. Our findings suggest that idioblast cells and laticifer cells have different biological roles in MIA biosynthesis and its regulation.
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Catharanthus , Catharanthus/metabolismo , Regulação da Expressão Gênica de Plantas , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Plântula/metabolismoRESUMO
Specialized metabolites are chemically complex small molecules with a myriad of biological functions. To investigate plant-specialized metabolite biosynthesis more effectively, we developed an improved method for virus-induced gene silencing (VIGS). We designed a plasmid that incorporates fragments of both the target gene and knockdown marker gene (phytoene desaturase, PDS), which identifies tissues that have been successfully silenced in planta. To demonstrate the utility of this method, we used the terpenoid indole alkaloid (TIA) pathway in Madagascar periwinkle (Catharanthus roseus) as a model system. Catharanthus roseus is a medicinal plant well known for producing many bioactive compounds, such as vinblastine and vincristine. Our VIGS method enabled the discovery of a previously unknown biosynthetic enzyme, serpentine synthase (SS). This enzyme is a cytochrome P450 (CYP) that produces the ß-carboline alkaloids serpentine and alstonine, compounds with strong blue autofluorescence and potential pharmacological activity. The discovery of this enzyme highlights the complexity of TIA biosynthesis and demonstrates the utility of this improved VIGS method for discovering unidentified metabolic enzymes in plants.
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Catharanthus/genética , Oxirredutases/metabolismo , Proteínas de Plantas/genética , Catharanthus/enzimologia , Catharanthus/metabolismo , Inativação Gênica , Genes de Plantas , Proteínas de Plantas/metabolismo , Alcaloides de Triptamina e Secologanina/metabolismo , Transdução de SinaisRESUMO
The enzymatic basis for quinine 1 biosynthesis was investigated. Transcriptomic data from the producing plant led to the discovery of three enzymes involved in the early and late steps of the pathway. A medium-chain alcohol dehydrogenase (CpDCS) and an esterase (CpDCE) yielded the biosynthetic intermediate dihydrocorynantheal 2 from strictosidine aglycone 3. Additionally, the discovery of an O-methyltransferase specific for 6'-hydroxycinchoninone 4 suggested the final step order to be cinchoninone 16/17 hydroxylation, methylation, and keto-reduction.
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Metiltransferases/metabolismo , Quinina/química , Quinina/metabolismo , Alcaloides de Vinca/química , Hidroxilação , Estrutura MolecularRESUMO
Purpose: Intranodal embolization using n-butyl cyanoacrylate glue is an emerging treatment option for persistent lymphatic leakage. This report describes the procedure details of intranodal embolization for groin lymphocele and evaluates the efficacy of intranodal embolization at our institution via retrospective chart review. Material and Methods: Nine consecutive patients (six men and three women; median age, 77.4 years; range, 43-95 years) who underwent intranodal embolization for groin lymphocele between January 2017 and December 2019 were included as study subjects. Intranodal lymphangiography with iodinated contrast was performed to confirm lymphatic leakage, followed by intranodal embolization using n-butyl cyanoacrylate glue mixed with iodized oil for all nine patients. The etiologies of lymphocele, technical and clinical success rates of intranodal embolization, duration of treatment, follow-up period, and acute and chronic complications were retrospectively investigated. Results: The etiologies of groin lymphoceles were the cutdown access of inguinal vessels (n = 7), lymph node biopsy (n = 1), and trauma (n = 1). The technical and clinical success rates of both lymphangiography and subsequent intranodal embolization were 100%. For intranodal embolization, 16.7%-33.3% n-butyl cyanoacrylate glue was used. The median duration of treatment was 2 days (range, 1-13 days). The follow-up period lasted 0-895 days (median, 9 days). No acute or chronic complications were observed. Conclusions: In this study, intranodal embolization showed promising results for groin lymphocele with a short duration of treatment with a median of 2 days. Intranodal embolization using n-butyl cyanoacrylate glue could be a treatment option for persistent groin lymphocele. However, further research is warranted to further evaluate the efficacy of intranodal embolization.
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Anomalous retro-psoas artery is an extremely rare congenital anomaly of the iliac arteries reported in the English-language literature. Using contrast-enhanced computed tomography, we incidentally diagnosed an anomalous retro-psoas artery in a 60-year-old male patient with right renal cancer. Although anomalous retro-psoas artery is extremely rare and mostly asymptomatic, knowledge of it is needed to avoid severe vascular complications in the event of vascular intervention and surgery.
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In Arabidopsis, the ASYMMETRIC LEAVES2 (AS2) protein plays a key role in the formation of flat symmetric leaves via direct repression of the abaxial gene ETT/ARF3. AS2 encodes a plant-specific nuclear protein that contains the AS2/LOB domain, which includes a zinc-finger (ZF) motif that is conserved in the AS2/LOB family. We have shown that AS2 binds to the coding DNA of ETT/ARF3, which requires the ZF motif. AS2 is co-localized with AS1 in perinucleolar bodies (AS2 bodies). To identify the amino acid signals in AS2 required for formation of AS2 bodies and function(s) in leaf formation, we constructed recombinant DNAs that encoded mutant AS2 proteins fused to yellow fluorescent protein. We examined the subcellular localization of these proteins in cells of cotyledons and leaf primordia of transgenic plants and cultured cells. The amino acid signals essential for formation of AS2 bodies were located within and adjacent to the ZF motif. Mutant AS2 that failed to form AS2 bodies also failed to rescue the as2-1 mutation. Our results suggest the importance of the formation of AS2 bodies and the nature of interactions of AS2 with its target DNA and nucleolar factors including NUCLEOLIN1. The partial overlap of AS2 bodies with perinucleolar chromocenters with condensed ribosomal RNA genes implies a correlation between AS2 bodies and the chromatin state. Patterns of AS2 bodies in cells during interphase and mitosis in leaf primordia were distinct from those in cultured cells, suggesting that the formation and distribution of AS2 bodies are developmentally modulated in plants.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Ligação a DNA/metabolismo , Fatores de Transcrição/metabolismo , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Cotilédone/genética , Cotilédone/crescimento & desenvolvimento , Proteínas de Ligação a DNA/genética , Mutação , Fenótipo , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , Domínios Proteicos , Fatores de Transcrição/genética , Dedos de ZincoRESUMO
Treatments for refractory glaucoma include trabeculectomy, in which a filtering bleb is created to reduce aqueous pressure. Mitomycin C (MMC) is often used as an adjuvant to reduce post-trabeculectomy bleb scarring and consequent failure. However, scarring sometimes still occurs. Thus, we searched for more effective trabeculectomy adjuvants with high-throughput screening (HTS) of a library of 1,165 off-patent drug compounds. This revealed that amsacrine (AMSA), a DNA topoisomerase II (TOP2) inhibitor, was the top candidate. Compared to MMC, rabbits that underwent trabeculectomy with 10% AMSA had lower IOP at 42, 56, and 70 days (P < 0.01 at all measurement points) and a higher bleb score at 28, 42, 56, and 70 days (P = < 0.01, 0.04, 0.04, and < 0.01, respectively). Compared to saline, rabbits that received 1% AMSA also had lower IOP and better bleb score at all time points, without a sharp drop in IOP just after surgery (all P < 0.01). Both effects were milder than MMC at 7 days (P = 0.02 and <0.01, respectively). Thus, this study showed that HTS may help identify new, promising uses for off-patent drugs. Furthermore, trabeculectomy with AMSA at a suitable concentration may improve the prognosis after trabeculectomy compared to MMC.
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Amsacrina , Implantes para Drenagem de Glaucoma , Glaucoma , Trabeculectomia , Animais , Humanos , Amsacrina/farmacologia , Callithrix , Cicatriz/prevenção & controle , Túnica Conjuntiva/efeitos dos fármacos , Modelos Animais de Doenças , DNA Topoisomerases Tipo II/efeitos dos fármacos , DNA Topoisomerases Tipo II/genética , Cirurgia Filtrante , Glaucoma/tratamento farmacológico , Glaucoma/patologia , Glaucoma/cirurgia , Implantes para Drenagem de Glaucoma/efeitos adversos , Pressão Intraocular/efeitos dos fármacos , Cultura Primária de Células , Inibidores da Topoisomerase II/farmacologia , Trabeculectomia/efeitos adversosRESUMO
Catharanthus roseus is a medicinal plant well known for producing bioactive compounds such as vinblastine and vincristine, which are classified as terpenoid indole alkaloids (TIAs). Although the leaves of this plant are the main source of these antitumour drugs, much remains unknown on how TIAs are biosynthesised from a central precursor, strictosidine, to various TIAs in planta. Here, we have succeeded in showing, for the first time in leaf tissue of C. roseus, cell-specific TIAs localisation and accumulation with 10 µm spatial resolution Imaging mass spectrometry (Imaging MS) and live single-cell mass spectrometry (single-cell MS). These metabolomic studies revealed that most TIA precursors (iridoids) are localised in the epidermal cells, but major TIAs including serpentine and vindoline are localised instead in idioblast cells. Interestingly, the central TIA intermediate strictosidine also accumulates in both epidermal and idioblast cells of C. roseus. Moreover, we also found that vindoline accumulation increases in laticifer cells as the leaf expands. These discoveries highlight the complexity of intercellular localisation in plant specialised metabolism.
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Catharanthus/citologia , Catharanthus/metabolismo , Metabolômica , Folhas de Planta/citologia , Alcaloides de Triptamina e Secologanina/metabolismo , Técnicas de Cultura de Células , Análise de Componente PrincipalRESUMO
PURPOSE: To investigate the potential of colchicine to improve bleb function after trabeculectomy. METHODS: To find the maximum usable colchicine concentration, an ocular irritation study was performed with the Draize test at concentrations of 0.001%, 0.01% and 0.1%. Additionally, the synergistic effect of topical colchicine instillation and MMC application to surgical site was evaluated in a rabbit model by measuring changes after trabeculectomy in intraocular pressure (IOP) and bleb morphology score at 3, 7, 14, 21, 28, 35, 42, and 49 days. RESULTS: Experiments with a rabbit model of trabeculectomy showed that 0.04% MMC plus 0.01% colchicine was more effective than saline and 0.04% MMC alone in maintaining IOP reduction at days 7-49 (P < 0.01 at all time points) and day 49 (P < 0.05), respectively, while 0.04% MMC alone was more effective than saline only at days 7-35 (P < 0.05 at all time points). 0.04% MMC plus 0.01% colchicine and 0.04% MMC alone were more effective than saline at preserving bleb score at days 7-21 and 35-49 (P < 0.05 at all time points) and at days 7-35 (P < 0.05 at all time points), respectively. CONCLUSION: Colchicine may be a promising adjuvant for strengthening the effect of MMC and improving the survival of the filtering bleb in trabeculectomy.
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Vesícula/tratamento farmacológico , Colchicina/uso terapêutico , Oftalmopatias/tratamento farmacológico , Mitomicina/uso terapêutico , Neovascularização Patológica/tratamento farmacológico , Trabeculectomia/métodos , Alquilantes/uso terapêutico , Animais , Vesícula/fisiopatologia , Vesícula/cirurgia , Quimioterapia Combinada , Oftalmopatias/cirurgia , Masculino , Neovascularização Patológica/cirurgia , Coelhos , Moduladores de Tubulina/uso terapêuticoRESUMO
Shoot phototropism benefits growth and metabolism in land plants by enabling them to position their photosynthetic organs in favorable light conditions. Nonvascular land plants, like the ancestors of modern mosses, are believed to have been among the first plants to occupy the land. To understand the evolutional history of shoot phototropism in land plants, we have established a system for experimentally studying phototropism in gametophores of the moss Physcomitrella patens. Here we will describe the key points in our system, including obtaining etiolated gametophores, the light sources used for inducing bending, and the methods for evaluation of phototropic responses.