TtCel7A: A Native Thermophilic Bifunctional Cellulose/Xylanase Exogluclanase from the Thermophilic Biomass-Degrading Fungus Thielavia terrestris Co3Bag1, and Its Application in Enzymatic Hydrolysis of Agroindustrial Derivatives.

The biomass-degrading thermophilic ascomycete fungus Thielavia terrestris Co3Bag1 produces TtCel7A, a native bifunctional cellulase/xylanase GH7 family. The purified TtCel7A, with an estimated molecular weight of 71 kDa, was biochemically characterized. TtCel7A displayed an optimal pH of 5.5 for both activities and an optimal temperature of 60 and 50 °C for cellulolytic and xylanolytic activities, respectively. The half-lives determined for cellulase activity were 140, 106, and 41 min at 50, 60, and 70 °C, respectively, whereas the half-lives observed for xylanase activity were 24, 10, and 1.4 h at 50, 60, and 70 °C, respectively. The KM and Vmax values were 3.12 mg/mL and 50 U/mg for cellulase activity and 0.17 mg/mL and 42.75 U/mg for xylanase activity. Circular dichroism analysis suggests changes in the secondary structure of TtCel7A in the presence of CMC as the substrate, whereas no modifications were observed with beechwood xylan. TtCel7A displayed the excellent capability to hydrolyze CMC, beechwood xylan, and complex substrates such as oat bran, wheat bran, and sugarcane bagasse, with glucose and cellobiose being the main products released; also, slightly less endo cellulase and xylanase activities were observed. Thus, suggesting TtCel7A has an exo- and endomode of action. Based on the characteristics of the enzyme, it might be considered a good candidate for industrial applications.

Development of a novel clinical decision support tool for diabetes prevention and feasibility of its implementation in primary care.

Prediabetes impacts 88 million U.S. adults, yet uptake of evidence-based treatment with intensive lifestyle interventions and metformin remains exceedingly low. After incorporating feedback from 15 primary care providers collected during semi-structured interviews, we developed a novel Prediabetes Clinical Decision Support (PreDM CDS) from August 2019 to February 2020. This tool included order options enabling prediabetes management in a single location within the electronic health record. We conducted a retrospective observational study examining the feasibility of implementing this tool at Erie Family Health Centers, a large community health center, examining its use and related outcomes among patients for whom it was used vs not. Overall, 7,424 eligible patients were seen during the implementation period (February 2020 to August 2021), and the PreDM CDS was used for 108 (1.5 %). Using the PreDM CDS was associated with higher rates of hemoglobin A1c orders (70.4 % vs 22.2 %; p < 0.001), lifestyle counseling (38.0 % vs 7.8 %; p < 0.001), and metformin prescription orders (5.6 % vs 2.6 %; p = 0.06). Exploratory analyses revealed small, nonsignificant weight loss among patients for whom the PreDM CDS was used. This study demonstrates the feasibility of developing and implementing the PreDM CDS in primary care. Its low use was likely related to not imposing an interruptive 'pop-up' alert, as well as major changes in workflows and clinical priorities during the Covid-19 pandemic. Use of the tool was associated with improved process outcomes. Future efforts with the PreDM CDS should follow standard CDS implementation processes that were not possible due to the Covid-19 pandemic.

Rahnella sp., a Dominant Symbiont of the Core Gut Bacteriome of Dendroctonus Species, Has Metabolic Capacity to Degrade Xylan by Bifunctional Xylanase-Ferulic Acid Esterase.

Rahnella sp. ChDrAdgB13 is a dominant member of the gut bacterial core of species of the genus Dendroctonus, which is one of the most destructive pine forest bark beetles. The objectives of this study were identified in Rahnella sp. ChDrAdgB13 genome the glycosyl hydrolase families involved in carbohydrate metabolism and specifically, the genes that participate in xylan hydrolysis, to determine the functionality of a putative endo-1,4-ß-D-xylanase, which results to be bifunctional xylanase-ferulic acid esterase called R13 Fae and characterize it biochemically. The carbohydrate-active enzyme prediction revealed 25 glycoside hydrolases, 20 glycosyl transferases, carbohydrate esterases, two auxiliary activities, one polysaccharide lyase, and one carbohydrate-binding module (CBM). The R13 Fae predicted showed high identity to the putative esterases and glycosyl hydrolases from Rahnella species and some members of the Yersiniaceae family. The r13 fae gene encodes 393 amino acids (43.5 kDa), containing a signal peptide, esterase catalytic domain, and CBM48. The R13 Fae modeling showed a higher binding affinity to ferulic acid, α-naphthyl acetate, and arabinoxylan, and a low affinity to starch. The R13 Fae recombinant protein showed activity on α-naphthyl acetate and xylan, but not on starch. This enzyme showed mesophilic characteristics, displaying its optimal activity at pH 6.0 and 25°C. The enzyme was stable at pH from 4.5 to 9.0, retaining nearly 66-71% of its original activity. The half-life of the enzyme was 23 days at 25°C. The enzyme was stable in the presence of metallic ions, except for Hg2+. The products of R13 Fae mediated hydrolysis of beechwood xylan were xylobiose and xylose, manifesting an exo-activity. The results suggest that Rahnella sp. ChDrAdgB13 hydrolyze xylan and its products could be assimilated by its host and other gut microbes as a nutritional source, demonstrating their functional role in the bacterial-insect interaction contributing to their fitness, development, and survival.

Hidrometrocolpos neonatal: a propósito de un caso de difícil diagnóstico / Neonatal hydrometrocolpos: a clinical case of difficult diagnosis

RESUMEN El diagnóstico de las malformaciones fetales sigue siendo un reto para el ginecoobstetra, a pesar de los avances en medicina fetal y en tecnología ultrasonográfica. El hidrometrocolpos es una dilatación del útero y vagina por el acúmulo de secreciones, producto de una obstrucción genital usualmente asociada a malformaciones congénitas. Presentamos el caso de una recién nacida con hidrometrocolpos congénito, cuya sospecha diagnóstica en los estudios pélvicos fue de quistes de ovario, pero se debía al incremento de una hidronefrosis bilateral. Al nacimiento, se decidió descomprimir quirúrgicamente la vía urinaria, momento en el que se encontró el hidrometrocolpos asociado a himen imperforado.

ABSTRACT The diagnosis of fetal malformations continues to be a challenge for the obstetriciangynecologist, despite advances in fetal medicine and ultrasonographic technology. Hydrometrocolpos is a dilatation of the uterus and vagina due to accumulation of secretions, product of genital obstruction usually associated with congenital malformations. We present the case of a female newborn with congenital hydrometrocolpos, whose diagnostic suspicion in pelvic studies was ovarian cysts, but it was due to the increase of bilateral hydronephrosis. At birth, it was decided to surgically decompress the urinary tract, at which time hydrometrocolpos associated with imperforate hymen was found.

Development and Evaluation of a Prediabetes Decision Aid in Primary Care: Examining Patient-Reported Outcomes by Language Preference and Educational Attainment.

PURPOSE: The purpose of this study was to examine the development and preliminary effectiveness of a novel Prediabetes Decision Aid on adoption of intensive lifestyle interventions (ILIs) and metformin. Little research has focused on increasing uptake of these evidence-based treatments, especially among non-English speakers and those with low educational attainment. METHODS: Investigators developed an English and Spanish decision aid displaying information about type 2 diabetes (T2DM) risk and treatments to prevent T2DM and prompting patients to identify next steps for management. This pilot study was a single-arm, pretest-posttest trial of 40 adult patients with prediabetes, obesity, and ≥1 office visit within the prior 12 months. Participants reviewed this tool briefly with a study team member, and data were collected on 3 coprimary outcomes: knowledge about T2DM risk, decisional conflict, and intention to adopt treatment. Exploratory outcomes included subsequent documentation of prediabetes in chart notes and adoption of ILIs or metformin. RESULTS: Almost all participants were women, with nearly half expressing Spanish language preference and low educational attainment. A nonsignificant increase in knowledge was observed across all subgroups. Decisional conflict was significantly reduced from pretest to posttest and was similar between subgroups defined by language preference and educational attainment. While intention to adopt ILIs increased across all subgroups, this change was only significant among Spanish speakers and participants with low educational attainment. At 6 months, 17 participants had subsequent provider documentation of prediabetes, and 12 adopted ILIs or metformin. CONCLUSIONS: The decision aid improved patient-reported outcomes and promoted treatment adoption in a diverse patient sample.

Control of microbial biofilm formation as an approach for biomaterials synthesis.

The search for new biomaterials with superior mechanical properties is the focus in the area of materials science. A promising pathway is drawing inspiration from nature to design and develop materials with enhanced properties. In this work, a novel strategy to produce functionalized supramolecular bionanomaterials from the microbial biofilm is reported. Tuneable biofilms with specific characteristics were obtained by controlling the culture condition of the microorganism. When the exopolysaccharide (EPS) production was desired the tryptone was the best nutritional component for the EPS production into the biofilm. However, for the expression of a high amount of amyloid protein the combination of peptone and glucose was the best nutritional choice. Each biofilm obtained showed its owner rheology properties. These properties were altered by the addition of extracellular DNA, which increased the viscosity of the biofilm and induced a viscoelastic hydrogel behavior. Besides, as a proof of concept of bionanomaterial, a novel supramolecular polymeric hybrid EPS-Amyloid protein (EPAP) was obtained from the biofilm and it was tested as a new natural functionalized support for enzyme immobilization. The results suggest that this technology could be used as a new concept to obtain biomaterials from biofilms by controlling the nutritional conditions of a microorganism. Understanding environmental factors affecting biofilm formation will help the development of methods for controlling biofilm production and therefore obtaining new biomaterials.

Tailoring doxorubicin sustainable release from biopolymeric smart matrix using congo red as molecular helper.

Doxorubicin (Dox) was co-encapsulated with congo red (CR) in order to increase drug encapsulation and sustain the release from gel microbeads composed of alginate-carboxy methyl guar gum (68/32) for oral controlled delivery. No release of either cargo molecule from the microbeads at pH 1.2 within 90 minutes was detected. However, 62% CR and 16% Dox were released from the gels at pH 7.4 at 37 °C in 8 hours when both the cargo molecules were studied alone. Presence of CR in the formulation reduces the release of Dox by about 25-30% under the same experimental conditions. Rheological properties of the formulations have been investigated at different temperatures between 20 and 37 °C. Shear thinning behavior was observed by steady-shear flow experiments for all formulations, and no yield stress was observed for any of the formulations. The temperature effect on Alg-CMGG-Dox-CR evidenced a synergic action between Dox and CR. Dynamic frequency sweep tests were performed to study the viscoelastic properties of the formulations. The patterns observed for Alg-CMGG indicated physical gel characteristics; however, all other formulations showed behaviour typical of concentrated solutions. These results confirm the interaction of Dox and CR, and the concomitant positive effect on sustainable release in oral delivery.

Binding and encapsulation of doxorubicin on smart pectin hydrogels for oral delivery.

Pectins (Pec) of 33 to 74 % esterification degree were tested with doxorubicin (Dox), a very high toxic drug widely used in cancer therapies. Pec with 35 and 55 % DE were selected because of the Dox binding higher than Pec microspheres of 35 and 55 % obtained by ionotropic gelation with Ca⁺² have 88 and 66 % Dox loading capacity. Kinetic Dox release showed more than 80.0 and about 30.0 % free drug from 35 % and 55 % Pec formulations at pH 7.4, and 37 °C after 1-h incubation, respectively. Besides, Dox release decrease to 12 % in 55 % Pec microsphere formulation after 1-year storage at 4 °C. FTIR analysis of Pec-Dox complex showed hipsochromic shifts for the σ(C=O), δ(N-H) and σ(C-C) vibrational modes compared to Dox spectrum suggesting strong interaction between the drug cargo and the matrix. Rheological studies of Pec and Pec-Dox samples flow behavior exhibited a shear-thinning nature. Fifty-five percent of Pec showed higher viscosity than the viscosity for 35 % Pec in all range of temperatures analyzed, and decreased when the temperature is raised. Besides, Pec-Dox complexes have higher viscosity values than those of the corresponding Pec samples, and viscosity curves as function of shear rate for 35 % Pec-Dox are above the curves of 55 % Pec-Dox. In both cases, the results are confirming significant interaction between the cargo and the matrix, which also was established in viscoelastic dynamic analysis.

Controlled release of sulfasalazine release from "smart" pectin gel microspheres under physiological simulated fluids.

Sulfasalazine (SLZ) is a synthetic nonsteroidal anti-inflammatory drug used mainly for the treatment of an inflammatory bowel and other diseases. Two pectins with different methylation degrees were blended to synthesized gel microspheres by ionotropic gelation for SLZ encapsulation. The encapsulation efficiency was found to be around of 99% in all formulations tested. However, different SLZ release profiles related to the methylation degrees of pectin were observed. Mixture of low methylated (LM) and high methylated (HM) pectins in the presence of calcium(II) displayed the best microsphere morphologies among the formulations tested determined by optical and electronic microscopies. The percentage of drug release using a mixture of LM and HM pectins after 255 min in simulated gastric fluid (pH = 1.2), simulated intestinal fluid (pH = 6.8), and phosphate buffer (pH = 7.4) were 15.0%, 47.0%, and 52.2%, respectively.

Focos de criptas aberrantes correlación endoscópico-morfológica con el uso de Endoscopios de alta resolución sin magnificación

Este trabajo tiene como propósito la descripción de los focos de criptas aberrantes utilizando colonoscopio de alta resolución sin magnificación. Los focos de criptas aberrantes se observan como lesiones planas o ligeramente elevadas, con un diámetro entre 1-4 mm, pueden presentar depresión central y con endoscopia de alta resolución, aún sin magnificación es posible ver el patrón de criptas. Usualmente se encuentran en la cercanía de adenomas. Una vez visualizada la lesión se procedió a realizar cromoscopía previa irrigación con 3-5 cc de ácido acético al 0,5 % como agente mucolítico, luego de 30 segundos se irrigó la zona con el colorante: azul de metileno al 0,5% o índigo carmín 0,5-1 %. Se intentó clasificar las lesiones de acuerdo a los criterios establecidos en la literatura para tratar de establecer correlación con el diagnóstico histológico. Se incluyeron 40 pacientes, en 67,5 % de los casos los focos de criptas aberrantes fueron localizados en colon recto-sigmoides. Existió una correlación endoscópica e histológica entre los focos de criptas aberrantes displásica y alta asociación con pólipos.

In our study, we used standard high resolution colonoscope to describe aberrant crypt foci. Aberrant crypt foci are either flat or slightly elevated lesions with a 1 to 4 mm of diameter with central depression. Usually these foci have been close to adenomas. Once the lesion was identified, it was first irrigated with 0.5% of acetic acid to remove mucus from the surface. After 30 seconds it was irrigated again with 0.5 % methylene blue or 0.5 to 1% indigo carmine. We tried to classify the lesion based on the literature criteria and compare it with the histopathology diagnosis. 40 patients were evaluated in this study, 67, 5 % of the aberrant crypt foci were located in rectosigmoid colon. We found direct histopathologic and endoscopic correlation between dysplasic aberrant crypt foci and high association with polyps.