Renal outcomes after contrast exposure in patients with diabetes who use sodium-glucose cotransporter 2 inhibitors.

BACKGROUND: Contrast-induced nephropathy has become increasingly prevalent as the age and prevalence of comorbidities in the general population have increased. Most cases of contrast-induced nephropathy are reversible; however, some may progress to acute kidney disease, and subsequently, to chronic kidney disease. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are known for their renoprotective effects. However, whether the use of these inhibitors affects the risk of contrast-induced kidney injury remains unclear. METHODS: Data were collected from the Taipei Medical University Clinical Research Database. We included patients with diabetes who had contrast exposure between 2016 and 2020 because of computed tomography or coronary angiography. The primary outcome was the risk of a major adverse kidney event (MAKE), which encompassed acute kidney disease, chronic kidney disease progression, and the need for renal replacement therapy. Overlap weighting was performed to reduce the effects of potential confounders. RESULTS: This study included 12 421 patients, who were divided into two groups: SGLT2i users (n = 920) and nonusers (n = 11 501). The follow-up period after contrast exposure was 6 months. The risk of a MAKE was lower in SGLT2i users than in nonusers (incidence, 36.9 vs. 49.9 per 1000 person-months, respectively; P = .0011). Furthermore, the incidence of acute kidney disease or chronic kidney disease progression was significantly lower in the SGLT2i users than in nonusers. However, no significant between-group difference was noted in the incidence of other MAKEs. CONCLUSIONS: SGLT2i may be safely used in diabetic patients needing contrast exposure. The risk of a MAKE may be lower in SGLT2i users than in nonusers.

A review on the hypoglycemic effect, mechanism and application development of natural dietary polysaccharides.

Diabetes mellitus (DM) as one chronic metabolic disease was greatly increased over recent decades. The major agents treating diabetes have noticeable side effects as well as the tolerability problems. The bioactive dietary polysaccharides from abundant natural resources exhibit good hypoglycemic effect with rare adverse effects, which might serve as a candidate to prevent and treat diabetes. However, the correlations between the hypoglycemic mechanism of polysaccharides and their structure were not mentioned in several studies, what's more, most of the current hypoglycemic studies on polysaccharides were based on in vitro and in vivo experiments, and there was a lack of knowledge about the effects in human clinical trials. The aim of this review is to discuss recent literature about the variety of dietary polysaccharides with hypoglycemic activity, as well the mechanism of action and the structure-function relationship are highlighted. Meanwhile, the application of dietary polysaccharides in functional foods and clinical medicine are realized with an in-depth understanding. So as to promote the exploration of dietary polysaccharides in low glycemic healthy foods or clinical medicine to prevent and treat diabetes.

A multiparametric clinic-ultrasomics nomogram for predicting extremity soft-tissue tumor malignancy: a combined retrospective and prospective bicentric study.

OBJECTIVE: We aimed at building and testing a multiparametric clinic-ultrasomics nomogram for prediction of malignant extremity soft-tissue tumors (ESTTs). MATERIALS AND METHODS: This combined retrospective and prospective bicentric study assessed the performance of the multiparametric clinic-ultrasomics nomogram to predict the malignancy of ESTTs, when compared with a conventional clinic-radiologic nomogram. A dataset of grayscale ultrasound (US), color Doppler flow imaging (CDFI), and elastography images for 209 ESTTs were retrospectively enrolled from one hospital, and divided into the training and validation cohorts. A multiparametric ultrasomics signature was built based on multimodal ultrasomic features extracted from the grayscale US, CDFI, and elastography images of ESTTs in the training cohort. Another conventional radiologic score was built based on multimodal US features as interpreted by two experienced radiologists. Two nomograms that integrated clinical risk factors and the multiparameter ultrasomics signature or conventional radiologic score were respectively developed. Performance of the two nomograms was validated in the retrospective validation cohort, and tested in a prospective dataset of 51 ESTTs from the second hospital. RESULTS: The multiparametric ultrasomics signature was built based on seven grayscale ultrasomic features, three CDFI ultrasomic features, and one elastography ultrasomic feature. The conventional radiologic score was built based on five multimodal US characteristics. Predictive performance of the multiparametric clinic-ultrasomics nomogram was superior to that of the conventional clinic-radiologic nomogram in the training (area under the receiver operating characteristic curve [AUC] 0.970 vs. 0.890, p = 0.006), validation (AUC: 0.946 vs. 0.828, p = 0.047) and test (AUC: 0.934 vs. 0.842, p = 0.040) cohorts, respectively. Decision curve analysis of combined training, validation and test cohorts revealed that the multiparametric clinic-ultrasomics nomogram had a higher overall net benefit than the conventional clinic-radiologic model. CONCLUSION: The multiparametric clinic-ultrasomics nomogram can accurately predict the malignancy of ESTTs.

Discovery of a Series of 5-Amide-1H-pyrazole-3-carboxyl Derivatives as Potent P2Y14R Antagonists with Anti-Inflammatory Characters.

UDPG/P2Y14R signaling pathway has been considered as a potential therapeutic target for innate immune system diseases. Based on the scaffold hopping strategy, a series of pyrazole analogues were designed and synthesized as novel P2Y14R antagonists with improved physicochemical properties, together with potential anti-inflammatory activities. Additionally, we designed and synthesized a fluorescent probe based on highly selective and potent PPTN to study the affinity of synthesized compounds. The optimized compound 16 (1-(4-fluorobenzyl)-5-(4-methylbenzamido)-1H-pyrazole-3-carboxylic acid, P2Y14R IC50 = 1.93 nM) showed strong binding ability to P2Y14R, high selectivity, notably improved solubility, and more favorable pharmacokinetic profiles. Moreover, compound 16 possessed extremely low cytotoxicity and anti-inflammatory effect in vitro. In an acute peritonitis model, compound 16 could effectively reduce the levels of inflammatory factor IL-6, IL-1ß, and TNF-α of mice induced by LPS. Compound 16, with potent in vitro and in vivo efficacy and favorable druggability, can be a promising candidate for further research.

Prevalence of IgE-mediated cow milk, egg, and peanut allergy in young Singapore children.

Background: The rising prevalence of food allergy reported in the United States, UK, and Australia may be attributable to the rise in peanut allergy prevalence. The food allergy prevalence in other parts of the world such as Asia is, however, less well documented. Objective: This study aimed to evaluate the prevalence of cow's milk, egg, and peanut allergies in a general population of Singaporean children below 30 months of age. Methods: A total of 4,115 children from the general population who attended well-baby visits between 2011 and 2015 completed standardized questionnaires to elicit a convincing history of food allergy to estimate the population prevalence of food allergies. Results: The prevalence of a convincing history of cow's milk allergy was 0.51% (95% confidence interval [CI], 0.3-0.7), hen's egg allergy 1.43% (95% CI, 1.1-1.8), and peanut allergy 0.27% (95% CI, 0.12-0.42). Of the 15 of 59 children with a convincing history of hen's egg allergy who consented, 12 (80%) had corroborative positive skin prick tests. Conclusion: The prevalence of food allergy, in particular peanut allergy, in children below 2 years of age is lower in this South East Asian population than reported in Western cohorts. Further research should focus on deciphering differential risk factors for food allergy across different geographical locations.

[The Expression and Clinical Correlation of the CD68 Factor in Acute Myeloid Leukemia Patients].

OBJECTIVE: To analyze the expression and clinical characteristics of CD68 in bone marrow and peripheral blood of patients with acute myeloid leukemia (AML). METHODS: The expression of CD68 in bone marrow blast cells was detected by four-color flow cytometry in 50 newly diagnosed AML patients and 23 controls. The expression of CD68 in peripheral blood of 85 newly diagnosed AML patients, 29 remission AML patients and 24 controls was detected by ELISA. The correlation between the expression rate of non-M3 AML bone marrow CD68, peripheral blood CD68 concentration and white blood cell count and other clinical data was compared respectively. RESULTS: The median CD68 expression rate in myeloid leukemia cells of non-M3 AML patients was 19.7%, significantly higher than control (0.2%) (P<0.001). The median concentration of non-M3 CD68 in peripheral blood was 67.97 pg/ml, significantly higher than in control (29.94 pg/ml)（P<0.01）. There was no statistically significant difference in the plasma CD68 concentration of the peripheral blood between the newly diagnosed (45.72 pg/ml) and the remission stage (55.12 pg/ml) of non-M3 AML patients by paired analysis (P>0.05). The results showed that the higher the expression rate of CD68 in bone marrow, the higher the count of white blood cells in peripheral blood, and the lower the count of hemoglobin and platelet in peripheral blood. The higher the plasma concentration of CD68 in peripheral blood, the higher the white blood cell count and the lower the complete remission rate. CONCLUSION: The expression of CD68 both in bone marrow and peripheral blood of patients with non-M3 AML is higher than that of control group. Patients with high expression of CD68 show a low rate of complete remission, suggesting that the expression level of CD68 is correlated with treatment response.

[Primary Diseases of 393 Cases of Pediatric Retinal Detachment and the Distribution of Clinical Characteristics].

OBJECTIVE: To investigate the primary diseases and the distribution of the clinical characteristics of pediatric retinal detachment. METHODS: Clinical records of patients aged 0-14 years old who had retinal detachment and who were hospitalized at the Department of Ophthalmology, West China Hospital, Sichuan University between January 2015 and December 2020, were retrospectively analyzed. The information on the demographic data, history, the scope of retinal detachment, and prognosis was retrieved and documented. RESULTS: A total of 464 eyes of 393 patients were included in this study, including 261 male and 132 female patients at a ratio of 1.98:1. The most common type of primary disease causing pediatric retinal detachments was associated with ocular dysplasia (227 cases, 57.8%), followed by trauma (78 cases, 19.8%) and myopia (56 cases, 14.2%). For infants and preschool children, the primary disease was predominantly ocular dysplasia at a rate of 81.8% (126 cases) and 55.8% (43 cases), respectively. For school-age children, in addition to ocular dysplasia (58 cases, 35.8%), myopia (49 cases, 30.2%) and ocular trauma (43 cases, 26.5%) also accounted for a large proportion of the primary diseases. The most common type of pediatric retinal detachment was rhegmatogenous retinal detachment (184 cases, 46.8%), and ocular trauma and myopia were the most common primary diseases, accounting for 37.5% (69/184) and 30.4% (56/184), respectively. 170 patients were diagnosed with traction retinal detachment, the second most common type of pediatric retinal detachment, and familial exudative vitreoretinopathy (FEVER) was the main primary disease, accounting for 47.6% (81/170). Exudative retinal detachment (39 cases, 9.9%) was the least common type of pediatric retinal detachment. Coats disease was the main primary disease causing exudative retinal detachment, accounting for 71.8% (28/39). After the first repair surgery, patients who had complete traction retinal detachment generally had poorer anatomical outcomes compared with those with complete rhegmatogenous retinal detachments (| Z|=3.026, P=0.002). The retinal break was most commonly seen on the temporal side in pediatric rhegmatogenous retinal detachments. In the retinal detachments caused by trauma, the most common type of retinal break was retinal tear, followed by ora serrata dialysis. Myopic rhegmatogenous retinal detachments were most commonly found in the round holes in lattice degeneration region. CONCLUSION: In the current study, boys were found to be more susceptible to retinal detachment than girls did. Ocular dysplasia, ocular trauma and myopia were major etiologic factors for pediatric retinal detachment. Appropriate information and education measures should be emphasized for different age groups.

Spiral-Dextrin Complex Crystals: Efficient Approach for Colon-Targeted Resveratrol Delivery.

In this work, spiral dextrin/resveratrol (SD/Res) crystal, a new colon-specific drug-delivery system, was established by a novel method of encapsulation and cocrystallization to improve the antidigestion ability compared with the SD/Res inclusion complex (SD/Res IC) prepared by encapsulation and coprecipitation. X-ray diffraction (XRD) and scanning electron microscopy (SEM) revealed that the SD/Res crystal formed a more regular and perfect crystallite than SD/Res IC. Moreover, the encapsulation ability and thermostability of the SD/Res crystal were enhanced as the chain length of SD was increased. In vitro digestion indicated that SD/Res IC merely achieved small intestine-targeted release of resveratrol, while the SD/Res crystal could act as a colon-specific delivery system to protect resveratrol from degradation by gastric acid and pancreatic enzymes. The SD-1/Res crystal presented much higher thermal stability and stronger gastrointestinal stability than other SD/Res crystals and SD/Res ICs, which facilitated its application as a novel colon-target delivery system for resveratrol.

Molecular Basis for Chemical Evolution of Flavones to Flavonols and Anthocyanins in Land Plants.

During the course of evolution of land plants, different classes of flavonoids, including flavonols and anthocyanins, sequentially emerged, facilitating adaptation to the harsh terrestrial environment. Flavanone 3ß-hydroxylase (F3H), an enzyme functioning in flavonol and anthocyanin biosynthesis and a member of the 2-oxoglutarate-dependent dioxygenase (2-ODD) family, catalyzes the hydroxylation of (2S)-flavanones to dihydroflavonols, but its origin and evolution remain elusive. Here, we demonstrate that functional flavone synthase Is (FNS Is) are widely distributed in the primitive land plants liverworts and evolutionarily connected to seed plant F3Hs. We identified and characterized a set of 2-ODD enzymes from several liverwort species and plants in various evolutionary clades of the plant kingdom. The bifunctional enzyme FNS I/F2H emerged in liverworts, and FNS I/F3H evolved in Physcomitrium (Physcomitrella) patens and Selaginella moellendorffii, suggesting that they represent the functional transition forms between canonical FNS Is and F3Hs. The functional transition from FNS Is to F3Hs provides a molecular basis for the chemical evolution of flavones to flavonols and anthocyanins, which contributes to the acquisition of a broader spectrum of flavonoids in seed plants and facilitates their adaptation to the terrestrial ecosystem.

[Establishment of Acquired Aplastic Anemia SD Rat Model].

OBJECTIVE: To establish a stable and rapidly rat model acquired aplastic anemia. METHODS: The SD rats were exposed to 137Csγ-ray at 3.5 and 4.0 Gy ( 91 cGy/min), and intraperitoneally injected with CTX at 35 mg/( kg·d) and CHL at 45 mg/( kg·d) in d 4, 6 and 8 after irradiation; the WBC, platelet and reticulocyte counts in peripheral blood, the smears and nucleated cells counts of bone marrow were observed. RESULTS: The levels of peripheral blood 3-lineage cells of SD rats treated with 137Csγ-irradiation combined with cyclophosphamide and chloramphenicol were significantly reduced, among which white blood cells, platelets and reticulocytes decreased rapidly, and the number of bone marrow nucleated cells decreased significantly; bone marrow pathological sections showed severe reduction of hematopoietic cells, and the non-hematopoietic cells such as fat cells increased, and a serve or lightly reduction of bone marrow cells were found. CONCLUSION: The rat model established by 137Csγ-ray irradiation combined with cyclophosphamide and chloramphenicol meets the clinical characteristics of aplastic anemia, and this study provides a stable rat model for the study of new therapeutic drugs for acquired aplastic anemia.

[Immunophenotypic Analysis of Patients with Adult Acute T-Lymphoblastic Leukemia].

OBJECTIVE: To analyze the results of immunophenotyping in patients with acute T-lymphoblastic leukemia (T-ALL) so as to improve the understanding of early T-cell precursor acute T-lymphoblastic leukemia (ETP-ALL) and near ETP-ALL with elevated CD5. METHODS: The results of immunophenotypes in 62 patients with newly diagnosed adult T-ALL in our hospital were retrospectively analyzed. According to the expression of CD5, CD1a, CD8 and My/S antigens, the patients were divided into 3 groups: ETP, near ETP and non-ETP. The immunophenotypic characteristics and basic clinical data of three groups were compared and analyzed. RESULTS: There were significant differences in age, white blood cell and plt counts as well as percentage of bone marrow blasts among three groups (P＜0.05). ETP and near ETP mainly belonged to Pro T and pre T, and non-ETP mainly belonged to cortical T and mature T. The positive rates of My/S, CD1a, CD8 and CD5 were significantly different among three groups (P＜0.01). The positive rates of CD4 and CD3 in non-ETP group were significantly higher than those in ETP and near-ETP group (P＜0.01). The positive rate of CD56 in near-ETP group was 50%, which was significantly higher than that in ETP group (16.7%) and non-ETP group (0%) (P＜0.01). CONCLUSION: According to the expression of CD5, CD1a, CD8 and My/S antigens, T-ALL patients can be divided into three groups: ETP, near ETP and non ETP, the immunophenotypes and basic clinical data among 3 groups are statistically different.

Effects of octenyl succinic anhydride groups distribution on the storage and shear stability of Pickering emulsions formulated by modified rice starch.

The modified rice starches by octenyl succinic anhydride (OS-RSs) with the same degree of substitution of 0.033 were prepared in aqueous solution using 3% or 20% (w/w) NaOH as a catalyst (OS-RS 3 or OS-RS 20). The results indicated that 20% of NaOH could destroy the structure of starch more seriously and led to more OS groups travelling to interior of starch granules. Furthermore, the storage stability and rheological properties of Pickering emulsions formulated by RS, OS-RS 3 and OS-RS 20 were investigated. Emulsion index of OS-RS 20 were always lower than OS-RS 3 during the storage time, while G', G" and viscosity of O/W Pickering emulsion formulated by OS-RS 20 particles were greater than those of OS-RS 3. OS-RS 3 with more hydrophobic groups in the granule surface could form more compact oil/water layers which contributed greater to the storage stability of Pickering emulsion due to the formation of more compact oil/water layers. However, OS-RS 20 with homogeneous distribution of OS groups in the whole particles resulted in the formation of a tighter droplet network structure that was more resistant to external impacts, which showed OS-RS 20 could improve the shear stability of Pickering emulsion.

A comparison study on polysaccharides extracted from Fructus Mori using different methods: structural characterization and glucose entrapment.

In this study, the structure characteristics and the hypoglycemic and antioxidant activities of mulberry fruit polysaccharides obtained by the commonly used hot water (MFPh)-, ultrasonic (MFPu)-, acid (MFPc)- and alkali (MFPa)-assisted extraction methods were investigated. NMR analysis indicated that the four polysaccharides had similar glycosidic linkage patterns. Scanning electron microscopy analyses showed that the surface morphology of the polysaccharides was greatly affected by the extraction methods. The results of the bioactivity assays indicated that MFPh exhibited stronger antioxidant and α-amylase inhibitory activities than the other polysaccharides. Moreover, all the polysaccharides showed good α-glucosidase inhibitory activities except for MFPu with the lowest molecular weight. These results suggested that acid, alkali, and ultrasonic-assisted extractions have different effects on the degradation of polysaccharides without changing the main structure compared with hot water extraction. In addition, the molecular weight of polysaccharides plays a key role in the bioactivity of the mulberry fruit polysaccharides.

Antidepressant-Like Effect and Mechanism of Action of Honokiol on the Mouse Lipopolysaccharide (LPS) Depression Model.

There is growing evidence that neuroinflammation is closely linked to depression. Honokiol, a biologically active substance extracted from Magnolia officinalis, which is widely used in traditional Chinese medicine, has been shown to exert significant anti-inflammatory effects and improve depression-like behavior caused by inflammation. However, the specific mechanism of action of this activity is still unclear. In this study, the lipopolysaccharide (LPS) mouse model was used to study the effect of honokiol on depression-like behavior induced by LPS in mice and its potential mechanism. A single administration of LPS (1 mg/kg, intraperitoneal injection) increased the immobility time in the forced swimming test (FST) and tail suspension test (TST), without affecting autonomous activity. Pretreatment with honokiol (10 mg/kg, oral administration) for 11 consecutive days significantly improved the immobility time of depressed mice in the FST and TST experiments. Moreover, honokiol ameliorated LPS-induced NF-κB activation in the hippocampus and significantly reduced the levels of the pro-inflammatory cytokines; tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and interferon γ (IFN-γ). In addition, honokiol inhibited LPS-induced indoleamine 2,3-dioxygenase (IDO) activation and quinolinic acid (a toxic product) increase and reduced the level of free calcium in brain tissue, thereby inhibiting calcium overload. In summary, our results indicate that the anti-depressant-like effects of honokiol are mediated by its anti-inflammatory effects. Honokiol may inhibit the LPS-induced neuroinflammatory response through the NF-κB signaling pathway, reducing the levels of related pro-inflammatory cytokines, and furthermore, this may affect tryptophan metabolism and increase neuroprotective metabolites.

[Establishment of 137Cs γ ray Combined with Cyclophosphamide and Chloramphenicol-Mediated Mouse Model of Acquired Aplastic Anemia].

OBJECTIVE: To improve and establish the mouse model with aplastic anemia (AA) mediated by 137Cs γ-ray irradiation combined with cyclophosphamide (CTX) and chloramphenicol (CHL) injection,so as to provide a stable model for studying the pathogenesis and treatment of AA . METHODS: The BALB/c mice were exposed to 137Cs γ-ray of 3-5 Gy(91 cGy/min) and were intraperitoneally injected with CTX of 25 mg/(kg.d) and CHL of 62.5 mg/(kg.d) at D 4,5 and 6 after irradiation; the WBC, platelet and reticulocyte counts in peripheral blood as well as the mucleated cell count in bone marrow and bone marrow smears were detected . RESULTS: The 3-lineage cells in peripheral blood of BALB/c mouse model with acquired AA were rapidly reduced, especially WBC, platelet and reticulocyte counts were lowest at D 14,the 3-lineage cells in peripheral blood were still severely reduced at D 28; the nucleated cell count in bone marrow significantly dcreased,the bone marrow hyperplasia was reduced or severely reduced; the pathological sections of bone marrow showed the severe reduction of hematopoietic cells and the increased of non-hematopoietic cells such as fat cells. CONCLUSION: The mouse model with acquired AA has been established by 137Cs γ-ray irradiation combined with CTX and CHL injection. All detection indicators of this model reach to diagnostic criteria for acquired AA,therefore this mouse model may be used as the model for study of pathogenesis and treatment of acquired AA.

Encapsulation of Vitamin E and Soy Isoflavone Using Spiral Dextrin: Comparative Structural Characterization, Release Kinetics, and Antioxidant Capacity during Simulated Gastrointestinal Tract.

Spiral dextrin subfraction (SD-40) obtained through enzyme debranching and gradient ethanol precipitation could interact with vitamin E (VE) or soy isoflavone (SIO) to form V-type inclusion complexes. The formation of two inclusion complexes was confirmed by Fourier transform-infrared spectroscopy, atomic force microscopy, and differential scanning calorimetry. In this study, an in vitro gastrointestinal model was used to investigate the breakdown of inclusion complexes and release behavior of bioactive compounds. The results indicated that the two inclusion complexes exhibited a controlled and sustained release behavior during digestion. In addition, the SD-40/VE inclusion complex presented higher stability and stronger antioxidant capacity than the SD-40/SIO inclusion complex. Furthermore, the first and zero order models were applied to understand the release kinetics of VE and SIO from inclusion complexes in the stomach, whereas the first order model was chosen to describe the release of VE and SIO from inclusion complexes in the intestine.

Biliary tract external drainage improves inflammatory mediators and pathomorphology of the intestine, liver, and lung in septic rats.

BACKGROUND: To investigate the effect of biliary tract external drainage (BTED) on inflammatory mediators and pathomorphism of intestine, liver, and lung in septic rats. METHOD: 48 SD rats (n = 8 per group) were randomized into six groups: control, sepsis, sepsis plus BTED, normal bile (obtained from eight healthy rats), and septic bile infusion for 6 hours respectively to test the effects of BTED bile infusion on cytokines' expression and tissue injury in the intestine, liver, and lung of septic/normal rats. Co-cultivation of intestinal epithelial cells (IEC-6) with bile for 12 hours was performed to evaluate the potential cytotoxicity of septic bile. Survival rate for sepsis plus BTED rats was detected compared with sepsis without BTED group (n = 20 per group) at 24, 48, and 72 hours, respectively. RESULTS: BTED for 6 hours significantly reduced the mRNA expression levels of tumor necrosis factor alpha (TNF-α) and IL-1ß (all p < 0.05 vs. sepsis group), whereas mRNA expression of TNF-α and IL-1ß in the intestine was increased after 6 hours' septic bile infusion compared with normal bile infusion group (all p < 0.05). TNF-α concentration in septic bile was significantly higher than that in the control group (p < 0.001). Tissue injury was significantly attenuated after 6 hours' BTED. CONCLUSIONS: BTED can significantly restrain the mRNA expression of TNF-α and IL-1ß in the intestine, liver, and lung and attenuate histological damage in septic rats.

Cloning and Functional Characterization of Two 4-Coumarate: CoA Ligase Genes from Selaginella moellendorffii.

Selaginella is an extant lycopodiophyte genus, which is representative of an ancient lineage of tracheophytes. The important evolutionary status makes it a valuable resource for the study of metabolic evolution in vascular plants. 4-coumarate: CoA ligase (4CL) is the pivotal enzyme that controls the flow of carbon through the phenylpropanoid metabolic pathway into the specific lignin, flavonoid, and wall-bound phenolics biosynthesis pathways. Although 4CLs have been extensively characterized in other vascular plants, little is known of their functions in Selaginella. Here, we isolated two 4CL genes (Sm4CL1 and Sm4CL2) from Selaginella moellendorffii. Based on the enzymatic activities of the recombinant proteins, both of these genes encoded bona fide 4CLs. The 4CL isoforms in S. moellendorffii have different activities: Sm4CL2 was more active than Sm4CL1. The enzymatic properties and gene expression patterns indicated that the 4CL genes have been conserved in the evolution of vascular plants.

Functional Characterization of a Hydroxyacid/Alcohol Hydroxycinnamoyl Transferase Produced by the Liverwort Marchantia emarginata.

The aerial organs of most terrestrial plants are covered by a hydrophobic protective cuticle. The main constituent of the cuticle is the lipid polyester cutin, which is composed of aliphatic and aromatic domains. The aliphatic component is a polyester between fatty acid/alcohol and hydroxycinnamoyl acid. The BAHD/HxxxD family enzymes are central to the synthesis of these polyesters. The nature of this class of enzymes in bryophytes has not been explored to date. Here, a gene encoding a fatty ω-hydroxyacid/fatty alcohol hydroxycinnamoyl transferase (HFT) has been isolated from the liverwort Marchantia emarginata and has been functionally characterized. Experiments based on recombinant protein showed that the enzyme uses ω-hydroxy fatty acids or primary alcohols as its acyl acceptor and various hydroxycinnamoyl-CoAs-preferentially feruloyl-CoA and caffeoyl-CoA-as acyl donors at least in vitro. The transient expression of a MeHFT-GFP fusion transgene in the Nicotiana benthamiana leaf demonstrated that MeHFT is directed to the cytoplasm, suggesting that the feruloylation of cutin monomers takes place there.