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Lanthanum carbonate (LC) is a phosphate binder used in end-stage renal disease (ESRD) with few adverse effects due to poor systemic absorption. Gastrointestinal deposition is likely due to alterations in epithelial permeability from inflammation in ESRD. It is challenging to detect in cases with minimal deposition and may be missed on endoscopy and biopsy. A 36-year-old with ESRD who was evaluated for gastrectomy was found to have LC deposition histologically. Years later, the excised portion had similar findings. This case allows for evaluation of LC gastropathy in a resection specimen, providing the opportunity to showcase its unique pathology features.
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INTRODUCTION: The COVID-19 pandemic led to visitor restrictions in many hospitals. Since care in the surgical intensive care unit (SICU) often engages visitors as surrogate decision-makers, we investigated whether there was an association between COVID-19-related visitor restrictions, goals of care discussions (GOCD), and patient outcomes in SICU patients. METHODS: We conducted a retrospective review of trauma and emergency general surgery (EGS) patients admitted to a rural tertiary SICU between July 2019 and April 2021, dividing patients into those admitted during COVID-19 visitor restrictions and those admitted at other times. Using univariate and multivariate logistic regression analyses, we compared the primary outcome, incidence of GOCD, and incidence of prolonged hospital (> 14 d) and intensive care unit length of stay (LOS, > 7 d) between the two groups. RESULTS: One hundred seventy nine of 368 study patients (48.6%) presented during restricted visitation. The proportion of GOCD was 38.0% and 36.5% in the restricted and nonrestricted visitation cohorts, respectively (P = 0.769). GOCD timing and outcomes were similar in both groups. The use of telecommunication increased during restricted visitation, as did the proportion of trauma patients admitted to the SICU. On multivariable logistic regression, age and patient category were independent predictors of GOCD. On outcomes analysis, visitor restriction was associated with prolonged hospital LOS for EGS patients (odds ratio 2.44, 95% confidence interval 1.01-5.91, P value 0.048). CONCLUSIONS: Restricted visitation was not associated with changes in frequency or outcome of GOCD, but was associated with prolonged hospital LOS among EGS patients who had SICU admissions. Further investigation of patient/surrogate satisfaction with virtual GOCD in the SICU setting is needed.
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COVID-19 , Cuidados Críticos , Humanos , Pandemias , Tempo de Internação , COVID-19/epidemiologia , Unidades de Terapia Intensiva , Planejamento de Assistência ao PacienteRESUMO
OBJECTIVE: Musculoskeletal discomfort is associated with repetitive movements and constrained body positions. The current meta-analysis was performed to determine the global prevalence of musculoskeletal symptoms among gynecologic surgeons who perform laparoscopy. METHODS: Sources included Embase, MEDLINE, PubMed, CINAHL, Web of Science Core Collection, Cochrane Central Register of Controlled Clinical Trials, and Google Scholar. Articles published between 1980 and 2022 were considered. Studies that assessed self-reported musculoskeletal symptoms were included. Relevant data were extracted and tabulated. RESULTS: Twelve studies met the inclusion criteria. In a pooled sample of 1619 surgeons, the estimated prevalence of musculoskeletal symptoms was 82% (95% confidence interval [CI], 70%-89%; I2 , 92%). Female sex was a risk factor, as identified by a pooled odds ratio of 4.64 (95% CI, 2.63-8.19; I2 , 0%) compared with male surgeons. Among surgeons who reported musculoskeletal symptoms, 30% (95% CI, 14%-52%; I2 , 95%) sought treatment and 3% (95% CI, 2%-6%; I2 , 0%) required work hour modifications. CONCLUSION: The current meta-analysis provides preliminary evidence of a high prevalence of musculoskeletal symptoms among gynecologic laparoscopic surgeons. Future research is needed to explore the underlying risk factors and interventional strategies to mitigate this risk.
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Laparoscopia , Dor Musculoesquelética , Doenças Profissionais , Cirurgiões , Humanos , Masculino , Feminino , Dor Musculoesquelética/etiologia , Dor Musculoesquelética/complicações , Doenças Profissionais/epidemiologia , Doenças Profissionais/etiologia , Prevalência , Ergonomia , Laparoscopia/efeitos adversosRESUMO
Lignin is a phenolic polymer deposited in the plant cell wall, and is mainly polymerized from three canonical monomers (monolignols), i.e. p-coumaryl, coniferyl and sinapyl alcohols. After polymerization, these alcohols form different lignin substructures. In dicotyledons, monolignols are biosynthesized from phenylalanine, an aromatic amino acid. Shikimate acts at two positions in the route to the lignin building blocks. It is part of the shikimate pathway that provides the precursor for the biosynthesis of phenylalanine, and is involved in the transesterification of p-coumaroyl-CoA to p-coumaroyl shikimate, one of the key steps in the biosynthesis of coniferyl and sinapyl alcohols. The shikimate residue in p-coumaroyl shikimate is released in later steps, and the resulting shikimate becomes available again for the biosynthesis of new p-coumaroyl shikimate molecules. In this study, we inhibited cytosolic shikimate recycling in transgenic hybrid aspen by accelerated phosphorylation of shikimate in the cytosol through expression of a bacterial shikimate kinase (SK). This expression elicited an increase in p-hydroxyphenyl units of lignin and, by contrast, a decrease in guaiacyl and syringyl units. Transgenic plants with high SK activity produced a lignin content comparable to that in wild-type plants, and had an increased processability via enzymatic saccharification. Although expression of many genes was altered in the transgenic plants, elevated SK activity did not exert a significant effect on the expression of the majority of genes responsible for lignin biosynthesis. The present results indicate that cytosolic shikimate recycling is crucial to the monomeric composition of lignin rather than for lignin content.
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Vias Biossintéticas , Lignina , Álcoois/metabolismo , Vias Biossintéticas/genética , Citosol/metabolismo , Lignina/metabolismo , Fenilalanina/metabolismo , Plantas Geneticamente Modificadas/metabolismoRESUMO
In sorghum (Sorghum bicolor) and other C4 grasses, brown midrib (bmr) mutants have long been associated with plants impaired in their ability to synthesize lignin. The brown midrib 30 (Bmr30) gene, identified using a bulk segregant analysis and next-generation sequencing, was determined to encode a chalcone isomerase (CHI). Two independent mutations within this gene confirmed that loss of its function was responsible for the brown leaf midrib phenotype and reduced lignin concentration. Loss of the Bmr30 gene function, as shown by histochemical staining of leaf midrib and stalk sections, resulted in altered cell wall composition. In the bmr30 mutants, CHI activity was drastically reduced, and the accumulation of total flavonoids and total anthocyanins was impaired, which is consistent with its function in flavonoid biosynthesis. The level of the flavone lignin monomer tricin was reduced 20-fold in the stem relative to wild type, and to undetectable levels in the leaf tissue of the mutants. The bmr30 mutant, therefore, harbors a mutation in a phenylpropanoid biosynthetic gene that is key to the interconnection between flavonoids and monolignols, both of which are utilized for lignin synthesis in the grasses.
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Endoplasmic reticulum (ER) stress is associated with acute kidney injury (AKI) caused by various mechanisms, including antibiotics, non-steroidal anti-inflammatory drugs, cisplatin, and radiocontrast. Tunicamycin (TM) is a nucleoside antibiotic that induces ER stress and is a commonly used model of AKI. 4-phenylbutyrate (4-PBA) is a chemical chaperone and histone deacetylase (HDAC) inhibitor and has been shown to protect the kidney from ER stress, apoptosis, and structural damage in a tunicamycin model of AKI. The renal protection provided by 4-PBA is attributed to its ability to prevent misfolded protein aggregation and inhibit ER stress; however, the HDAC inhibitor effects of 4-PBA have not been examined in the TM-induced model of AKI. As such, the main objective of this study was to determine if histone hyperacetylation provides any protective effects against TM-mediated AKI. The FDA-approved HDAC inhibitor vorinostat was used, as it has no ER stress inhibitory effects and therefore the histone hyperacetylation properties alone could be investigated. In vitro work demonstrated that vorinostat inhibited histone deacetylation in cultured proximal tubular cells but did not prevent ER stress or protein aggregation induced by TM. Vorinostat induced a significant increase in cell death, and exacerbated TM-mediated total cell death and apoptotic cell death. Wild type male mice were treated with TM (0.5 mg/kg, intraperitoneal injection), with or without vorinostat (50 mg/kg/day) or 4-PBA (1 g/kg/day). Mice treated with 4-PBA or vorinostat exhibited similar levels of histone hyperacetylation. Expression of the pro-apoptotic protein CHOP was induced with TM, and not inhibited by vorinostat. Further, vorinostat did not prevent any renal damage or decline in renal function caused by tunicamycin. These data suggest that the protective mechanisms found by 4-PBA are primarily due to its molecular chaperone properties, and the HDAC inhibitors used did not provide any protection against renal injury caused by ER stress.
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Injúria Renal Aguda , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Tunicamicina/efeitos adversos , Vorinostat/farmacologia , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/prevenção & controle , Animais , Linhagem Celular , Modelos Animais de Doenças , Masculino , Camundongos , Agregação Patológica de Proteínas/induzido quimicamente , Agregação Patológica de Proteínas/metabolismo , Agregação Patológica de Proteínas/patologia , Agregação Patológica de Proteínas/prevenção & controle , Tunicamicina/farmacologiaRESUMO
Lignocellulose, the structural component of plant cells, is a major agricultural byproduct and the most abundant terrestrial source of biopolymers on Earth. The complex and insoluble nature of lignocellulose limits its conversion into value-added commodities, and currently, efficient transformation requires expensive pretreatments and high loadings of enzymes. Here, we report on a fungus from the Parascedosporium genus, isolated from a wheat-straw composting community, that secretes a large and diverse array of carbohydrate-active enzymes (CAZymes) when grown on lignocellulosic substrates. We describe an oxidase activity that cleaves the major ß-ether units in lignin, thereby releasing the flavonoid tricin from monocot lignin and enhancing the digestion of lignocellulose by polysaccharidase mixtures. We show that the enzyme, which holds potential for the biorefining industry, is widely distributed among lignocellulose-degrading fungi from the Sordariomycetes phylum.
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Ascomicetos/enzimologia , Biopolímeros/química , Enzimas/química , Lignina/química , Ascomicetos/química , Biopolímeros/metabolismo , Enzimas/genética , Flavonoides/química , Lignina/metabolismo , Oxirredução , Oxirredutases/química , Oxirredutases/genética , Oxigenases/química , Especificidade por Substrato/genética , Triticum/enzimologia , Triticum/microbiologiaRESUMO
Invited for this month's cover is the research team from the D.O.E. Great Lake Bioenergy Research Center (GLBRC) at the University of Wisconsin-Madison. The cover image shows how a diverse team with expertise in many different fields works together in an integrated fashion to address complex problems. Only when the whole system, from field to the liquid fuels and co-products, is assessed, can we identify the key parameters needed to design an economically viable biorefinery-based economy. Cover art by Chelsea Mamott. The Full Paper itself is available at 10.1002/cssc.201903345.
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Parede Celular/metabolismo , Lignina/metabolismo , Morus/metabolismo , Parede Celular/genética , Cromatografia Gasosa-Espectrometria de Massas , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Estrutura Molecular , Morus/genética , MutaçãoRESUMO
The hydroxycinnamic acids p-coumaric acid (pCA) and ferulic acid (FA) add diversity to the portfolio of products produced by using grass-fed lignocellulosic biorefineries. The level of lignin-bound pCA in Zea mays was modified by the alteration of p-coumaroyl-CoA monolignol transferase expression. The biomass was processed in a lab-scale alkaline-pretreatment biorefinery process and the data were used for a baseline technoeconomic analysis to determine where to direct future research efforts to couple plant design to biomass utilization processes. It is concluded that future plant engineering efforts should focus on strategies that ramp up accumulation of one type of hydroxycinnamate (pCA or FA) predominantly and suppress that of the other. Technoeconomic analysis indicates that target extraction titers of one hydroxycinnamic acid need to be >50â g kg-1 biomass, at least five times higher than observed titers for the impure pCA/FA product mixture from wild-type maize. The technical challenge for process engineers is to develop a viable process that requires more than 80 % reduction of the isolation costs.
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RATIONALE: Self-esteem is an adaptive personality factor that has been associated with good physical health. While research has observed that self-esteem and physical health typically decline in older adulthood, there is a paucity of research investigating the associations between changes in self-esteem and physical health across the adult lifespan. OBJECTIVE: The present study examined whether changes in selfesteem and chronic disease exert reciprocal effects on subsequent changes in self-esteem and disease. In addition, it investigated whether individuals' age would moderate these associations. METHODS: The study analyzed data from 14,117 adult (18+) Canadians who completed surveys over 16 years, from cycles 1 to 9 of the National Population Health Survey (NPHS). Self-esteem, chronic diseases, and demographic information were collected. RESULTS: Cross-lagged panel analyses indicated reciprocal age-related associations between changes in self-esteem and chronic disease. Initial decline in self-esteem predicted subsequent increases in chronic disease, and initial increases in chronic disease predicted subsequent declines in self-esteem, only among young adults, and not middle-aged or older adults. CONCLUSION: These results suggest that age may qualify the associations between declines in self-esteem and physical health and that adverse changes in both factors may be particularly problematic for young adults' prospective personality functioning and physical health.
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Envelhecimento/psicologia , Doença Crônica/psicologia , Autoimagem , Adaptação Psicológica , Adulto , Fatores Etários , Idoso , Canadá , Doença Crônica/tendências , Feminino , Humanos , Entrevistas como Assunto/métodos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Pesquisa Qualitativa , Inquéritos e QuestionáriosRESUMO
Syringyl (S) lignin content and the syringyl-to-guaiacyl (S/G) lignin ratio are important characteristics of wood and lignocellulosic biomass. Although numerous methods are available for estimating S lignin units and the S/G ratio, in this work, a new method based on Raman spectroscopy that uses the 370 cm-1 Raman band-area intensity (370-area) was developed. The reliability of the Raman approach for determining S content was first tested by the quantitative analysis of three syringyl lignin models by sampling them, separately, in dioxane and in Avicel. Good linear correlations between the 370 cm-1 intensity and model concentrations were obtained. Next, the percent syringyl (%S) lignin units in various woods were measured by correlating the 370 cm-1 Raman intensity data with values of S units in lignin determined by three regularly used methods, namely, thioacidolysis, DFRC, and 2D-HSQC NMR. The former two methods take into account only the monomers cleaved from ß-O-4-linked lignin units, whereas the NMR method reports S content on the whole cell wall lignin. When the 370-area intensities and %S values from the regularly used methods were correlated, good linear correlations were obtained ( R2 = 0.767, 0.731, and 0.804, respectively, for the three methods). The correlation with the highest R2, i.e., with the 2D NMR method, is proposed for estimating S units in wood lignins by Raman spectroscopy as, in principle, both represent the whole cell wall lignin and not just the portion of lignin that gets cleaved to release monomers. The Raman analysis method is quick, uses minimal harmful chemicals, is carried out nondestructively, and is insensitive to the wet or dry state of the sample. The only limitations are that the sample of wood contains at least 30% S and not be significantly fluorescent, although the latter can be mitigated in some cases.
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Lignina/química , Análise Espectral Raman/métodos , Madeira/química , Estrutura Molecular , Árvores/químicaRESUMO
BACKGROUND: Consumption of added sugars (AS) and sugar-sweetened beverages (SSB) may adversely affect adolescents' weight and cardiovascular disease risk. Reliance on self-reported dietary assessment methods is a common research limitation, which could be overcome by dietary intake biomarkers. AIM: The investigation was a proof-of-concept study to evaluate the proposed carbon isotope ratio (δ13C) biomarker of AS intake in adolescents, using a controlled feeding design. METHODS: Participants (n = 33, age 15.3 years, 53% female) underwent two seven-day controlled feeding periods in a randomly assigned order. Diets were matched in composition except for AS content (5% or 25% of total energy). Fasting fingerstick blood samples were collected daily during each diet period. RESULTS: Fingerstick δ13C values changed from day 1 to 8 by -0.05 ± 0.071 on 5% AS, and +0.03 ± 0.083 on 25% AS (p ≤ 0.001). Reliability was demonstrated between day 7 and 8 δ13C values on the 5% (ICC = 0.996, p ≤ 0.001) and 25% (ICC = 0.997, p ≤ 0.001) AS diets. CONCLUSIONS: Larger scale investigations are warranted to determine if this technique could be applied to population-level research in order to help assess the effectiveness of interventions aimed at reducing the consumption of AS or SSB intake.
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Isótopos de Carbono/sangue , Dieta/métodos , Açúcares da Dieta/sangue , Açúcares da Dieta/farmacologia , Adolescente , Biomarcadores/sangue , Feminino , Humanos , Masculino , Reprodutibilidade dos Testes , TempoRESUMO
BACKGROUND: High added sugar (AS) intake is associated with obesity and poor diet quality. Guidelines recommended limiting AS to 5-10% of total energy intake, but palatability and feasibility of this AS intake level is uncertain. OBJECTIVE: To compare adolescents' perceptions of hunger, fullness, and palatability in response to a low AS adequate fiber (LASAF; 5% total energy from AS and 13.5â¯g fiber/1000â¯kcal) and a high AS low fiber (HASLF; 25% total energy form AS and 8.2â¯g/1000â¯kcal) diet. DESIGN: Adolescents (nâ¯=â¯32, age: 15.3⯱â¯1.6â¯yrs., BMI percentile: 47⯱â¯4, 15 male) completed a randomized, crossover, controlled feeding study. Participants consumed calorie-matched LASAF and HASLF diets for 7â¯days, separated by a 4â¯week washout. Body weight was monitored daily on each diet. Hunger, fullness, and palatability were assessed via 100â¯mm visual analogue scales at the end of each feeding period. Differences were assessed with paired sample t-tests. Data are expressed as mean⯱â¯SD. RESULTS: Participants remained weight stable, and no difference in weight change between diet conditions was detected (LASAF: -0.06⯱â¯0.7 vs. HASLF: -0.02⯱â¯0.6â¯kg, pâ¯=â¯0.751). Less hunger (LASAF: 24.1⯱â¯14.6 vs. HASLF: 32.1⯱â¯17.6â¯mm, pâ¯=â¯0.024) and greater fullness (LASAF: 70.2⯱â¯12.3 vs. HASLF: 61.3⯱â¯18.1â¯mm, pâ¯=â¯0.006) were reported on the LASAF compared to the HASLF. Participants reported the diets to be equally palatable (LASAF: 39.6⯱â¯12.9 vs. HASLF: 37.2⯱â¯17.8â¯mm, pâ¯=â¯0.440). CONCLUSIONS: Adolescents perceive a LASAF diet to be as palatable as a HASLF diet, but more satiating. LASAF diets should be investigated as a strategy for weight control in adolescents.
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Dieta/psicologia , Fibras na Dieta/administração & dosagem , Açúcares da Dieta/administração & dosagem , Saciação , Adolescente , Estudos Cross-Over , Dieta/estatística & dados numéricos , Feminino , Humanos , Masculino , Projetos PilotoRESUMO
Background: Objective indicators of dietary intake (e.g., biomarkers) are needed to overcome the limitations of self-reported dietary intake assessment methods in adolescents. To our knowledge, no controlled feeding studies to date have evaluated the validity of urinary sodium, nitrogen, or sugar excretion as dietary biomarkers in adolescents.Objective: This investigation aimed to evaluate the validity of urinary sodium, nitrogen, and total sugars (TS) excretion as biomarkers for sodium, protein, and added sugars (AS) intake in nonobese adolescents.Methods: In a crossover controlled feeding study design, 33 adolescents [12-18 y of age, 47 ± 25th percentile (mean ± SD) of body mass index (BMI; in kg/m2) for age] consumed 5% AS [low added sugars (LAS)] and 25% AS [high added sugars (HAS)] isocaloric, macronutrient-matched (55% carbohydrate, 30% fat, and 15% protein) diets for 7 d each, in a randomly assigned order, with a 4-wk washout period between diets. On the final 2 d of each diet period, 24-h urine samples were collected. Thirty-two adolescents completed all measurements (97% retention).Results: Urinary sodium was not different from the expected 90% recovery (mean ± SD: 88% ± 18%, P = 0.50). Urinary nitrogen was correlated with protein intake (r = 0.69, P < 0.001), although it was below the 80% expected recovery (62% ± 7%, P < 0.001). Urinary TS values were correlated with AS intake during the HAS diet (r = 0.77, P < 0.001) and had a higher R2 value of 0.28 than did AS intake (R2 = 0.36). TS excretion differed between LAS (0.226 ± 0.09 mg/d) and HAS (0.365 ± 0.16 mg/d) feeding periods (P < 0.001).Conclusions: Urinary sodium appears to be a valid biomarker for sodium intake in nonobese adolescents. Urinary nitrogen is associated with protein intake, but nitrogen excretion rates were less than previously reported for adults, possibly owing to adolescent growth rates. TS excretion reflects AS at 25% AS intake and was responsive to the change in AS intake. Thus, urinary biomarkers are promising objective indicators of dietary intake in adolescents, although larger-scale feeding trials are needed to confirm these findings. This trial was registered at clinicaltrials.gov as NCT02455388.
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Carboidratos/urina , Carboidratos da Dieta/administração & dosagem , Proteínas Alimentares/administração & dosagem , Nitrogênio/urina , Sódio na Dieta/administração & dosagem , Sódio/urina , Adolescente , Biomarcadores , Criança , Estudos Cross-Over , Feminino , Humanos , MasculinoRESUMO
Hematopoietic stem/progenitor cells (HSPCs) are formed during ontogeny from hemogenic endothelium in the ventral wall of the dorsal aorta (VDA). Critically, the cellular mechanism(s) allowing HSPC egress and migration to secondary niches are incompletely understood. Matrix metalloproteinases (MMPs) are inflammation-responsive proteins that regulate extracellular matrix (ECM) remodeling, cellular interactions, and signaling. Here, inhibition of vascular-associated Mmp2 function caused accumulation of fibronectin-rich ECM, retention of runx1/cmyb+ HSPCs in the VDA, and delayed caudal hematopoietic tissue (CHT) colonization; these defects were absent in fibronectin mutants, indicating that Mmp2 facilitates endothelial-to-hematopoietic transition via ECM remodeling. In contrast, Mmp9 was dispensable for HSPC budding, being instead required for proper colonization of secondary niches. Significantly, these migration defects were mimicked by overexpression and blocked by knockdown of C-X-C motif chemokine-12 (cxcl12), suggesting that Mmp9 controls CHT homeostasis through chemokine regulation. Our findings indicate Mmp2 and Mmp9 play distinct but complementary roles in developmental HSPC production and migration.
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Movimento Celular , Células-Tronco Embrionárias/citologia , Células-Tronco Hematopoéticas/citologia , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Nicho de Células-Tronco , Animais , Proliferação de Células , Quimiocina CXCL12/metabolismo , Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Células-Tronco Embrionárias/metabolismo , Células-Tronco Embrionárias/fisiologia , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Hematopoéticas/fisiologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/metabolismoRESUMO
Lignin is a major polymer in the secondary plant cell wall and composed of hydrophobic interlinked hydroxyphenylpropanoid units. The presence of lignin hampers conversion of plant biomass into biofuels; plants with modified lignin are therefore being investigated for increased digestibility. The bacterium Sphingomonas paucimobilis produces lignin-degrading enzymes including LigD, LigF and LigG involved in cleaving the most abundant lignin interunit linkage, the ß-aryl ether bond. In this study, we expressed the LigD, LigF and LigG (LigDFG) genes in Arabidopsis thaliana to introduce postlignification modifications into the lignin structure. The three enzymes were targeted to the secretory pathway. Phenolic metabolite profiling and 2D HSQC NMR of the transgenic lines showed an increase in oxidized guaiacyl and syringyl units without concomitant increase in oxidized ß-aryl ether units, showing lignin bond cleavage. Saccharification yield increased significantly in transgenic lines expressing LigDFG, showing the applicability of our approach. Additional new information on substrate specificity of the LigDFG enzymes is also provided.