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INTRODUCTION: With the increased knowledge about the physiology of liposuction, procedures that involve larger body surface area are now being performed, which means an increased risk of hemorrhage and the need for transfusion. Homologous blood transfusion poses well-known risks, prompting the use of alternative and safer techniques such as autologous donation. To demonstrate the utility of autologousOBJECTIVE: transfusion in a cohort of patients undergoing large volume liposuction. Observational, retrospective andMETHODS: longitudinal cohort study in patients undergoing liposuction, in the period 2017-2021. Autologous donation was performed one month and two weeks prior to surgery, ensuring that all patients were prescribed medication and prophylactic dietary measures to avoid hemoglobin depletion. 40 les of women between 26 and 46 years of age, with a bodyRESULTS AND CONCLUSIONS: mass index between 24 and 30 m/kg2 and a median aspirated volume of 6.3 Lt were analyzed. Quantied bleeding averaged 675 cc. Both initial hemoglobin and hematocrit had statistically signicant differences with values in the postoperative period (pn all. However, vital signs remained stable and no transfusion-associated adverse events occurred. Autologous transfusion is an excellent strategy for the management of bleeding in large volume liposuction.

Objective: To describe the case of a patient with modeling disease and overview the literature. Case description: 62-year-old female patient with a history of injection with modeling substances 20 years ago presenting late onset reaction to these and ASIA syndrome, managed with surgical debridement, negative pressure system and skin graft placement. Conclusion: Complications due to the use of biopolymers can be severe and their management must be multidisciplinary. Keywords: modeling disease, biopolymers, skin grafting.

NMR spectroscopy has recently been utilized to determine the absolute amounts of organic molecules with metrological traceability since signal intensity is directly proportional to the number of each nucleus in a molecule. The NMR methodology that uses hydrogen nucleus (1H) to quantify chemicals is called quantitative 1H-NMR (1H qNMR). The quantitative method using 1H qNMR for determining the purity or content of chemicals has been adopted into some compendial guidelines and official standards. However, there are still few reports in the literature regarding validation of 1H qNMR methodology. Here, we coordinated an international collaborative study to validate a 1H qNMR based on the use of an internal calibration methodology. Thirteen laboratories participated in this study, and the purities of three samples were individually measured using 1H qNMR method. The three samples were all certified via conventional primary methods of measurement, such as butyl p-hydroxybenzoate Japanese Pharmacopeia (JP) reference standard certified by mass balance; benzoic acid certified reference material (CRM) certified by coulometric titration; fludioxonil CRM certified by a combination of freezing point depression method and 1H qNMR. For each sample, 1H qNMR experiments were optimized before quantitative analysis. The results showed that the measured values of each sample were equivalent to the corresponding reference labeled value. Furthermore, assessment of these 1H qNMR data using the normalized error, En-value, concluded that statistically 1H qNMR has the competence to obtain the same quantification performance and accuracy as the conventional primary methods of measurement.

Introduction: The growing of maize (zea mays L.) in Latin America is carried out under the intensive production system, which causes loss of soil fertility due to excessive nitrogen fertilizer. Objective: Analyzing the response of maize var. Jala to genera of plant growth promoting bacteria (PGPB) endophytes of maize var. Mexicana (teocintle) when using nitrogen fertilizer such as urea at 50 % under greenhouse and soil conditions. Methodology: A randomized block design was established with five treatments, three controls and twenty repetitions. The treatments with inoculated maize seeds and 50 % urea were: Burkholderia (KO) and Burkholderia sp. 14 (B14); the controls were: non-inoculated maize with 50 % urea (RC1), non-inoculated maize irrigated with water (FC) and non-inoculated maize with 100 % urea (RC2). The experiments were established under greenhouse and soil conditions. The response variables were radical dry weight (RDW) and fresh weight of grain (FWG). ANOVA and Tukey’s mean comparison (P ≤ 0.01) were carried out. Results: In both environments, treatment B14 showed statistically higher values than those obtained with FC (full control), but statistically equal to RC2 (relative control). Study limitations: Maize varieties have specific and beneficial endophytic PGPB genera, which are not necessarily for regional varieties. Originality: PGPB endophytes from teocintle (ancestor of maize) were used because of their close genetic relationship with the maize var. Jala. Conclusion: PGPB endophytes from teocintle are a viable option for sustainable maize production.

Teratological diagnosis in polyplacophorans has mostly been based on primary structure (Chiton scleritome) with four types (hypermerism, hypomerism, coalescence and splitting), including some deformities in plates; however, diagnoses are hardly detailed for each sclerite. As part of a monthly research campaign conducted by a year investigating Chiton articulatus, we found several cases with a complex combination of abnormalities (chiton scleritome + sclerites) that complement the previous classification. Identification and diagnosis of abnormalities were assigned in terms of sclerite number, number of slits per side in the insertion area of intermediate sclerites and the merging grade between sclerites. Slit and slit-ray numbers were baseline to identify combination of abnormalities; the slit formula of Chiton articulatus is 13–18[I]/1[II-VII]/16–22[VIII]. We found four abnormality cases with combinations among them: a) perfect hypomerism, b) heterogeneous coalescence, c) homogeneous coalescence, d) imperfect hypermerism. Heterogeneous coalescence was most common abnormality, with a different merging degree between sclerites and with specimens showing combinations of abnormal states such as: normality/coalescence, coalescence/hypomerism, coalescence/hypermerism and normality/hypermerism; so these complex abnormalities can generates asymmetry between the right and left side of a chiton. We postulate that coalescence is an intermediate step toward perfect hypomerism or perfect hypermerism. The width/length ratio suggests that an abnormal chiton increases its width to compensate and retain a normal body appearance despite different degrees of abnormal development.

The first records of the pelagic cephalopod Argonauta nouryi nbsp Lorois captured alive in shallow waters of the Central Mexican Pacific are presented complementing such information with a brief analysis of specimens of Argonauta cornuta nbsp Linnaeus from stomach contents of commercial fish caught in the sport fishing of said region

A shift scheduling problem for a call centre is solved by applying a column generation method. The objective is to find an employee schedule within the planning horizon that meets staffing requirements provided by the customer and that minimises the costs of excess-staffing and under-staffing. To solve the problem, two iterative models were used: the first being a mixed integer-linear programming model, the master problem, and the second being a column generation model. In a first approach, the master problem was solved considering a fixed number of shift patterns. The second approach was to use both models iteratively to generate shift patterns to be used in the master problem. Both approaches met all constraints relatively fast, but the column generation model decreased the cost considerably when cheaper shift patterns were generated. The models contribute to the current literature by providing flexibility to consider under-coverage and over-coverage at two levels.

The increased interest in using monoclonal antibodies (mAbs) as a platform for biopharmaceuticals has led to the need for new analytical techniques that can precisely assess physicochemical properties of these large and very complex drugs for the purpose of correctly identifying quality attributes (QA). One QA, higher order structure (HOS), is unique to biopharmaceuticals and essential for establishing consistency in biopharmaceutical manufacturing, detecting process-related variations from manufacturing changes and establishing comparability between biologic products. To address this measurement challenge, two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) methods were introduced that allow for the precise atomic-level comparison of the HOS between two proteins, including mAbs. Here, an inter-laboratory comparison involving 26 industrial, government and academic laboratories worldwide was performed as a benchmark using the NISTmAb, from the National Institute of Standards and Technology (NIST), to facilitate the translation of the 2D-NMR method into routine use for biopharmaceutical product development. Two-dimensional 1H,15N and 1H,13C NMR spectra were acquired with harmonized experimental protocols on the unlabeled Fab domain and a uniformly enriched-15N, 20%-13C-enriched system suitability sample derived from the NISTmAb. Chemometric analyses from over 400 spectral maps acquired on 39 different NMR spectrometers ranging from 500 MHz to 900 MHz demonstrate spectral fingerprints that are fit-for-purpose for the assessment of HOS. The 2D-NMR method is shown to provide the measurement reliability needed to move the technique from an emerging technology to a harmonized, routine measurement that can be generally applied with great confidence to high precision assessments of the HOS of mAb-based biotherapeutics.

Resolution enhancement for glutamate (Glu), glutamine (Gln) and glutathione (GSH) in the human brain by TE-optimized point-resolved spectroscopy (PRESS) at 7 T is reported. Sub-TE dependences of the multiplets of Glu, Gln, GSH, γ-aminobutyric acid (GABA) and N-acetylaspartate (NAA) at 2.2–2.6 ppm were investigated with density matrix simulations, incorporating three-dimensional volume localization. The numerical simulations indicated that the C4-proton multiplets can be completely separated with (TE1, TE2) = (37, 63) ms, as a result of a narrowing of the multiplets and suppression of the NAA 2.5 ppm signal. Phantom experiments reproduced the signal yield and lineshape from simulations within experimental errors. In vivo tests of optimized PRESS were conducted on the prefrontal cortex of six healthy volunteers. In spectral fitting by LCModel, Cramer–Rao lower bounds (CRLBs) of Glu, Gln and GSH were 2 ± 1, 5 ± 1 and 6 ± 2 (mean ± SD), respectively. To evaluate the performance of the optimized PRESS method under identical experimental conditions, stimulated-echo spectra were acquired with (TE, TM) = (14, 37) and (74, 68) ms. The CRLB of Glu was similar between PRESS and short-TE stimulated-echo acquisition mode (STEAM), but the CRLBs of Gln and GSH were lower in PRESS than in both STEAM acquisitions. Copyright © 2010 John Wiley & Sons, Ltd.

Transcription of the ferric citrate import system is regulated by ferric citrate binding to the outer membrane transporter FecA. A signal indicating transporter occupancy is relayed across the outer membrane to energy-transducing and regulatory proteins embedded in the cytoplasmic membrane. Because transcriptional activation is not coupled to ferric citrate import, an allosteric mechanism underlies this complex signaling mechanism. Using evolution-based statistical analysis we have identified a sparse but structurally connected network of residues that links distant functional sites in FecA. Functional analyses of these positions confirm their involvement in the mechanism that regulates transcriptional activation in response to ferric citrate binding at the cell surface. This mechanism appears to be conserved and provides the structural basis for the allosteric signaling of TonB-dependent transporters.

In the preceding communication we described a fully synthetic pathway to the structure proposed for ( )-diazonamide A (1, Scheme 1).[1] This material is not identical to the natural product, which raises the obvious question: What is the true structure of diazonamide A? Herein we provide an answer. In addition, we report that defined synthetic entity 8 induces, with equal potency, a toxic phenotype in cell culture indistinguishable from that produced by natural ( )-diazonamide A. Spectroscopic data for the heterocyclic cores of diazonamides A and B are nearly identical.[2] So when the crystal structure of a p-bromobenzamide derivative of diazonamide B was reported as 3,[2a] the diazonamide A assignment Scheme 1. Initial diazonamide structure assignments.

Thiomorpholine-N-borane was synthesized via amine displacement of BH3 from tetrahydrofuran–borane which had been prepared from NaBH4 and BF3·Et2O in tetrahydrofuran. Acid-catalyzed hydrolysis occurs only slightly faster than for morpholine–borane, the difference being attributed to a small difference in the electronic inductive effects of sulfur and oxygen in the 4-position of the respective adducts. Reaction with NaOCl exhibits a stoichiometric [OCl−]:[S(CH2)4NHBH3] ratio of 5:1. This is attributed to consumption of 3 mol of hypochlorite for the oxidation of hydridic hydrogen in BH3, one for the chlorination of nitrogen and another in attack at sulfur presumably resulting in sulfoxide formation. At pH 9.1–10.4, the initial reaction of hypochlorite with thiomorpholine–borane is several times faster than with morpholine–borane and, unlike the reaction with morpholine–borane, relatively insensitive to pH. Whereas hypochlorite oxidation of morpholine–borane has been proposed to occur primarily through reaction with HOCl, it is speculated that thiomorpholine–borane is also susceptible to attack by hypochlorite ion.

The conformational analysis of a series of menthone and isomenthone imines was accomplished by 1HNMR spectroscopy and was compared with molecular modeling computations. The (E) menthone derivatives contain about 20% of a 2,5-diaxial chair conformer at room temperature (22–23°C), whereas in the (E)- and (Z)-isomenthone counterparts the major conformer has axial isopropyl and equatorial methyl groups. These peculiar conformations are a consequence of pseudo-allylic A(1,3) interactions between the isopropyl group and the non-bonded pair of electrons of the nitrogen, which dominate the axial–axial interactions between the alkyl chains and the protons of the ring in the isomenthone imines and are of considerable importance in the menthone derivatives. Copyright © 1999 John Wiley & Sons, Ltd.

trans-2-Isopropyl-5-methylcyclohexanone (menthone) may be considered as locked in the diequatorial conformer. However, the cis isomer (isomenthone) may exist in two major conformations with the isopropyl group either equatorial or axial. No measure of the equilibrium mix has been determined previously. Proton NMR allows such a determination at room temperature via the vicinal couplings between protons on carbons 5 and 6. Combined with modelling results, the calculated value with the isopropyl axial was 79%, of the total in good agreement with an earlier prediction. Two different conformer searching programs were employed to determine the various conformer populations. It was found that molecular mechanics estimates of the Boltzmann distributions varied considerably from the experimental results. These differences were not due to substituent electronegativity effects or to hydrogen bonding with the deuterochloroform solvent. Semiempirical PM3 and low-level ab initio calculations of the conformer energies also failed to account adequately for the conformer populations. DFT calculations (Becke3LYP/6–31G*) with zero-point energy corrections gave results most consistent with the experimentally determined values (67.5% axial isopropyl conformer). © 1988 John Wiley & Sons, Ltd.

In this work, we applied nuclear magnetic resonance (NMR) spectroscopy to rapidly assess higher order structure (HOS) comparability in protein samples. Using a variation of the NMR fingerprinting approach described by Panjwani et al. [2010. J Pharm Sci 99(8):3334–3342], three nonglycosylated proteins spanning a molecular weight range of 6.5–67 kDa were analyzed. A simple statistical method termed easy comparability of HOS by NMR (ECHOS-NMR) was developed. In this method, HOS similarity between two samples is measured via the correlation coefficient derived from linear regression analysis of binned NMR spectra. Applications of this method include HOS comparability assessment during new product development, manufacturing process changes, supplier changes, next-generation products, and the development of biosimilars to name just a few. We foresee ECHOS-NMR becoming a routine technique applied to comparability exercises used to complement data from other analytical techniques.

In the preceding communication we described a fully synthetic pathway to the structure proposed for ( )-diazonamide A (1, Scheme 1).[1] This material is not identical to the natural product, which raises the obvious question: What is the true structure of diazonamide A? Herein we provide an answer. In addition, we report that defined synthetic entity 8 induces, with equal potency, a toxic phenotype in cell culture indistinguishable from that produced by natural ( )-diazonamide A. Spectroscopic data for the heterocyclic cores of diazonamides A and B are nearly identical.[2] So when the crystal structure of a p-bromobenzamide derivative of diazonamide B was reported as 3,[2a] the diazonamide A assignment Scheme 1. Initial diazonamide structure assignments.

The aryl hydrocarbon receptor nuclear translocator (ARNT) serves as the obligate heterodimeric partner for bHLH-PAS proteins involved in sensing and coordinating transcriptional responses to xenobiotics, hypoxia, and developmental pathways. Although its C-terminal transactivation domain is dispensable for transcriptional activation in vivo, ARNT has recently been shown to use its N-terminal bHLH and/or PAS domains to interact with several transcriptional coactivators that are required for transcriptional initiation after xenobiotic or hypoxic cues. Here we show that ARNT uses a single PAS domain to interact with two coiled coil coactivators, TRIP230 and CoCoA. Both coactivators interact with the same interface on the ARNT PAS-B domain, located on the opposite side of the domain used to associate with the analogous PAS domain on its heterodimeric bHLH-PAS partner HIF-2α. Using NMR and biochemical studies, we identified the ARNT-interacting motif of one coactivator, TRIP230 as an LXXLL-like nuclear receptor box. Mutation of this motif and proximal sequences disrupts the interaction with ARNT PAS-B. Identification of this ARNT-coactivator interface illustrates how ARNT PAS-B is used to form critical interactions with both bHLH-PAS partners and coactivators that are required for transcriptional responses.

Selenium has significant health benefits, including potent cancer prevention activity and roles in immune function and the male reproductive system. Selenium-containing proteins, which incorporate this essential micronutrient as selenocysteine, are proposed to mediate the positive effects of dietary selenium. Presented here are the solution NMR structures of the selenoprotein SelM and an ortholog of the selenoprotein Sep15. These data reveal that Sep15 and SelM are structural homologs that establish a new thioredoxin-like protein family. The location of the active-site redox motifs within the fold together with the observed localized conformational changes after thiol-disulfide exchange and measured redox potential indicate that they have redox activity. In mammals, Sep15 expression is regulated by dietary selenium, and either decreased or increased expression of this selenoprotein alters redox homeostasis. A physiological role for Sep15 and SelM as thiol-disulfide oxidoreductases and their contribution to the quality control pathways of the endoplasmic reticulum are discussed.

We have used NMR spectroscopy to characterize an oligonucleotide stem loop structure based on the pre-element of an oncogenic microRNA, miR-21. This predicted stem-loop structure is cleaved from the precursor of miR-21 (pre-miR-21) by the nuclease Dicer. It is also a critical feature recognized by the protein complex that converts the primary transcript (pri-miR-21) into the pre-miRNA. The secondary structure of the native sequence is poorly defined by NMR due to rapid exchange of imino protons with solvent; however, replacement of two adjacent putative G•U base pairs with G•C base pairs retains the conformation of the hairpin observed by chemical probing and stabilizes it sufficiently to observe most of the imino proton resonances of the molecule. The observed resonances are consistent with the predicted secondary structure. In addition, a peak due to a loop uridine suggests an interaction between it and a bulged uridine in the stem. Assignment of non-exchangeable proton resonances and characterization of NOEs and coupling constants allows inference of the following features of the structure: extrahelicity of a bulged adenosine, deviation from A-form geometry in a base-paired stem, and consecutive stacking of the adenosines in the 5' side of the loop, the guanosine of the closing base pair, and a cross-strand adenosine. Modeling of the structure by restrained molecular dynamics suggests a basis for the interaction between the loop uridine, the bulged uridine in the stem, and an A•U base pair in the stem.

In order to study bioavailability of lysine of two soybeans meals (PSA and PSB) for broilers with different ureaseactivity (UA of 0.11 and 0.17 units of pH change), an experiment was performed. Two hundred and ten (210) chicksRoss were used from 1 to 21 d of age, in a completely randomized design with 7 treatments with three replicateseach one. Treatments were: 1) Basal diet sorghum-soybean meal (deficient in lysine), 2) Basal diet+0.05% L-lysine,3) Basal diet+0.10% L-lysine, 4) Basal diet+0.05% lysine from PSA, 5) Basal diet+0.10% lysine from PSA, 6) Basaldiet+0.05% lysine from PSB and 7) Basal diet+0.10% lysine from PSB. The growth results obtained were explainedby multiple linear regression equation, based on the data of weight gain, consumption of synthetic lysine and intakesof lysine from soybean meals: Y=X1+ Y= 375.419 + 0.0378 X1 + 0.0366 X2 + 0.0376 X3; where X1 accountedsupplementation with L-lysine, X2 with PSA (with addition of raw hull) and X3 with PSB (with addition of cockedhull). Comparison of the PSA or PSB slopes with the L-lysine slope (i.e. 100 % bioavailability) showed them all tohave >95 % lysine availability: PS A= 97 %, PSB= 99 %. These results indicate that UA of soybean meal affectedthe bioavailability of lysine; therefore more than 0.12 urcase activity units of pH change, lower is the availability.

El presente estudio tuvo como propósito investigar si la necesidad de treonina en la dieta no se incrementa al someter a pollos de engorda a un calendario cerrado de vacunación, manejo que representa mayor estrés. Se realizó un experimento con 990 pollos de engorda Arbor Acres, mixtos, de 1 a 42 días de edad, utilizando seis tratamientos con cinco repeticiones de 33 pollos cada uno. Se utilizó un diseño completamente al azar con un arreglo factorial 3 × 2; tres niveles de treonina en función del requerimiento informado por el NRC 1994 (95%, 100% y 105%) y dos programas de vacunación [abierto: Dos vacunas de Newcastle y cerrado; dos vacunas de Newcastle, una de infección de la Bolsa de Fabricio (IBF) y una bacterina de E. coli]. Se emplearon dietas en base a sorgo más pasta de soya para iniciación y finalización, con L-treonina para lograr los niveles requeridos. Los resultados en 42 días para ganancia de peso, consumo de alimento, conversión alimentaria, mortalidad general y resultados séricos de los títulos de anticuerpos por la prueba de inhibición de la hemoaglutinación para la enfermedad de Newcastle, no indicaron diferencias estadísticas (P 0.05) entre tratamientos. Los resultados de este estudio sugieren que las necesidades de treonina al 95% de lo que señala el NRC en 1994, son adecuadas para óptimo comportamiento productivo e inmunidad de pollos de engorda.