Chemoproteomic discovery of a covalent allosteric inhibitor of WRN helicase.

WRN helicase is a promising target for treatment of cancers with microsatellite instability (MSI) due to its essential role in resolving deleterious non-canonical DNA structures that accumulate in cells with faulty mismatch repair mechanisms1-5. Currently there are no approved drugs directly targeting human DNA or RNA helicases, in part owing to the challenging nature of developing potent and selective compounds to this class of proteins. Here we describe the chemoproteomics-enabled discovery of a clinical-stage, covalent allosteric inhibitor of WRN, VVD-133214. This compound selectively engages a cysteine (C727) located in a region of the helicase domain subject to interdomain movement during DNA unwinding. VVD-133214 binds WRN protein cooperatively with nucleotide and stabilizes compact conformations lacking the dynamic flexibility necessary for proper helicase function, resulting in widespread double-stranded DNA breaks, nuclear swelling and cell death in MSI-high (MSI-H), but not in microsatellite-stable, cells. The compound was well tolerated in mice and led to robust tumour regression in multiple MSI-H colorectal cancer cell lines and patient-derived xenograft models. Our work shows an allosteric approach for inhibition of WRN function that circumvents competition from an endogenous ATP cofactor in cancer cells, and designates VVD-133214 as a promising drug candidate for patients with MSI-H cancers.

Assessment of the antinociceptive effect of a single fentanyl bolus dose in children: A pharmacokinetic and pharmacodynamic analysis based on the nociception level index during sevoflurane general anesthesia.

BACKGROUND: The Nociception Level Index has shown benefits in estimating the nociception/antinociception balance in adults, but there is limited evidence in the pediatric population. Evaluating the index performance in children might provide valuable insights to guide opioid administration. AIMS: To evaluate the Nociception Level Index ability to identify a standardized nociceptive stimulus and the analgesic effect of a fentanyl bolus. Additionally, to characterize the pharmacokinetic/pharmacodynamic relationship of fentanyl with the Nociception Level Index response during sevoflurane anesthesia. METHODS: Nineteen children, 5.3 (4.1-6.7) years, scheduled for lower abdominal or urological surgery, were studied. After sevoflurane anesthesia and caudal block, a tetanic stimulus (50 Hz, 60 mA, 5 s) was performed in the forearm. Following the administration of fentanyl 2 µg/kg intravenous bolus, three similar consecutive tetanic stimuli were performed at 5-, 15-, and 30-min post-fentanyl administration. Changes in the Nociception Level Index, heart rate, mean arterial pressure, and bispectral index were compared in response to the tetanic stimuli. Fentanyl plasma concentrations and the Nociception Level Index data were used to elaborate a pharmacokinetic/pharmacodynamic model using a sequential modeling approach in NONMEM®. RESULTS: After the first tetanic stimulus, both the Nociception Level Index and the heart rate increased compared to baseline (8 ± 7 vs. 19 ± 10; mean difference (CI95) -12(-18--6) and 100 ± 10 vs. 102 ± 10; -2(-4--0.1)) and decrease following fentanyl administration (19 ± 10 vs. 8 ± 8; 12 (5-18) and 102 ± 10 vs. 91 ± 11; 11 (7-16)). In subsequent tetanic stimuli, heart rate remained unchanged, while the Nociception Level Index progressively increased within 15 min to values similar to those before fentanyl. An allometric weight-scaled, 3-compartment model best characterized the pharmacokinetic profile of fentanyl. The pharmacokinetic/pharmacodynamic modeling analysis revealed hysteresis between fentanyl plasma concentrations and the Nociception Level Index response, characterized by plasma effect-site equilibration half-time of 1.69 (0.4-2.9) min. The estimated fentanyl C50 was 1.93 (0.73-4.2) ng/mL. CONCLUSION: The Nociception Level Index showed superior capability compared to traditional hemodynamic variables in discriminating different nociception-antinociception levels during varying fentanyl concentrations in children under sevoflurane anesthesia.

Short-term high-light intensity and low temperature improve the quality and flavor of lettuce grown in plant factory.

BACKGROUND: Lettuce holds a prominent position in the year-round supply of vegetables, offering a rich array of health-beneficial substances, such as dietary fiber, phenolic compounds, lactucopicrin and lactucin. As such, its flavor has garnered increasing attention. Balancing the enhancement of beneficial compounds with the reduction of undesirable taste is a key focus of scientific research. To investigate short-term management to improve the nutritional quality and flavor of lettuce, combinations of different light intensities (200, 500 and 800 µm ol m-2 s-1) and temperatures (10 and 22 °C) were applied separately to 'Lollo Rosso' and 'Little Butter Lettuce' for 7 days before harvest. RESULTS: The results obtained showed that increasing light intensity at low temperatures decreased nitrate content and increased soluble sugar, soluble protein, anthocyanin and phenolic compound content. In the case of lettuce flavor, the bitterness-related metabolites such as lactucin and lactucopicrin were reduced with high light intensity at a low temperature of 10 °C. With this combination, the fructose and glucose contents increased, significantly improving lettuce flavor. CONCLUSION: Higher light intensity combined with low temperature for 7 days before harvest effectively improved the nutritional quality and flavor of lettuce, suggesting its great potential for use in horticultural practices. © 2024 Society of Chemical Industry.

Sequential Collinear Photofragmentation and Atomic Absorption Spectroscopy for Online Laser Monitoring of Triatomic Metal Species.

Industrial chemical processes are struggling with adverse effects, such as corrosion and deposition, caused by gaseous alkali and heavy metal species. Mitigation of these problems requires novel monitoring concepts that provide information on gas-phase chemistry. However, selective optical online monitoring of the most problematic diatomic and triatomic species is challenging due to overlapping spectral features. In this work, a selective, all-optical, in situ gas-phase monitoring technique for triatomic molecules containing metallic atoms was developed and demonstrated with detection of PbCl2. Sequential collinear photofragmentation and atomic absorption spectroscopy (CPFAAS) enables determination of the triatomic PbCl2 concentration through detection of released Pb atoms after two consecutive photofragmentation processes. Absorption cross-sections of PbCl2, PbCl, and Pb were determined experimentally in a laboratory-scale reactor to enable calibration-free quantitative determination of the precursor molecule concentration in an arbitrary environment. Limit of detection for PbCl2 in the laboratory reactor was determined to be 0.25 ppm. Furthermore, the method was introduced for in situ monitoring of PbCl2 concentration in a 120 MWth power plant using demolition wood as its main fuel. In addition to industrial applications, the method can provide information on chemical reaction kinetics of the intermediate species that can be utilized in reaction simulations.

Recently differentiated epimastigotes from Trypanosoma cruzi are infective to the mammalian host.

Trypanosoma cruzi, the etiologic agent of Chagas disease, has a complex life cycle in which four distinct developmental forms alternate between the insect vector and the mammalian host. It is assumed that replicating epimastigotes present in the insect gut are not infective to mammalian host, a paradigm corroborated by the widely acknowledged fact that only this stage is susceptible to the complement system. In the present work, we establish a T. cruzi in vitro and in vivo epimastigogenesis model to analyze the biological aspects of recently differentiated epimastigotes (rdEpi). We show that both trypomastigote stages of T. cruzi (cell-derived and metacyclic) are able to transform into epimastigotes (processes termed primary and secondary epimastigogenesis, respectively) and that rdEpi have striking properties in comparison to long-term cultured epimastigotes: resistance to complement-mediated lysis and both in vitro (cell culture) and in vivo (mouse) infectivity. Proteomics analysis of all T. cruzi stages reveled a cluster of proteins that were up-regulated only in rdEpi (including ABC transporters and ERO1), suggesting a role for them in rdEpi virulence. The present work introduces a new experimental model for the study of host-parasite interactions, showing that rdEpi can be infective to the mammalian host.

Chemical elemental analysis of single acoustic-levitated water droplets by laser-induced breakdown spectroscopy.

Laser-induced breakdown spectroscopy is presented for trace element detection of liquid samples by analyzing a single droplet levitated by ultrasonic waves. A single liquid droplet is placed in the node of a standing acoustic wave produced by a uniaxial levitator for further chemical analysis. The acoustic levitator consists of a commercial Langevin-type transducer, attached to a concave mechanical amplifier, and a concave reflector. A micro-syringe was used to manually place individual liquid droplet samples in the acoustic levitation system. For chemical analysis, a laser-induced plasma is produced by focusing a single laser pulse on the levitated water droplet after it partially dries. The performance of the acoustic levitator on micron-sized droplets is discussed, and the detection of Ba, Cd, Hg, and Pb at parts per million (milligrams/liter) and sub-parts per million levels is reported. The process, starting from placing the sample in the acoustic levitator and ending on the chemical identification of the traces, takes a few minutes. The approach is particularly interesting in applications demanding limited volumes of liquid samples and relative simple and inexpensive techniques.

Propofol pharmacokinetic and pharmacodynamic profile and its electroencephalographic interaction with remifentanil in children.

BACKGROUND: Propofol and remifentanil are commonly combined during total intravenous anesthesia. The impact of remifentanil in this relationship is poorly quantified in children. Derivation of an integrated pharmacokinetic and pharmacodynamic propofol model, containing remifentanil pharmacodynamic interaction information, enables propofol effect-site target-controlled infusion in children with a better prediction of its hypnotic effect when both drugs are combined. AIMS: We designed this study to derive an integrated propofol pharmacokinetic-pharmacodynamic model in children and to describe the pharmacodynamic interaction between propofol and remifentanil on the electroencephalographic bispectral index effect. METHODS: Thirty children (mean age: 5.45 years, range 1.3-11.9; mean weight: 23.5 kg, range 8.5-61) scheduled for elective surgery with general anesthesia were studied. After sevoflurane induction, maintenance of anesthesia was based on propofol and remifentanil. Blood samples to measure propofol concentration were collected during anesthesia maintenance and up to 6 hours in the postoperative period. Bispectral index data were continuously recorded throughout the study. A pharmacokinetic-pharmacodynamic model was developed using population modeling. The Greco model was used to examine the pharmacokinetic-pharmacodynamic interaction between propofol and remifentanil for BIS response RESULTS: Propofol pharmacokinetic data from a previous study in 53 children were pooled with current data and simultaneously analyzed. Propofol pharmacokinetics were adequately described by a three-compartment distribution model with first-order elimination. Theory-based allometric relationships based on TBW improved the model fit. The Greco model supported an additive interaction between propofol and remifentanil. Remifentanil showed only a minor effect in BIS response. CONCLUSION: We have developed an integrated propofol pharmacokinetic-pharmacodynamic model that can describe the pharmacodynamic interaction between propofol and remifentanil for BIS response. An additive interaction was supported by our modeling analysis.

Enhanced self-mixing interferometry based on volume Bragg gratings and laser diodes emitting at 405-nm wavelengths.

Self-mixing interferometry (SMI) represents a robust, self-aligned technique for metrology applications. Recently, it has been shown that the detection of the frequency-modulated (FM) signal enhances the conventional SMI signal based on the amplitude modulation. Here, an all-optical, simple and effective alternative approach to detect the FM self-mixing signal is presented. We demonstrate the enhanced self-mixing approach using a laser diode emitting at 405 nm and a volume Bragg grating (VBG) to map frequency to intensity modulations for further optical detection. Our approach overcomes the spectral range limitation of the edge filtering approach based on molecular absorption filters, since the VBGs can be fabricated at any spectral range.

Detection of single microparticles in airflows by edge-filter enhanced self-mixing interferometry.

A laser Doppler velocimetry (LDV) sensor using the edge-filter enhanced self-mixing interferometry (ESMI) is presented based on speed measurements of single microparticles. The ESMI detection utilizes an acetylene edge-filter that maps the frequency modulation of a semiconductor laser into an intensity modulation as the laser wavelength is tuned to the steep edge of the absorption profile. In this work, the ESMI signal was analyzed for aerosol particles of different sizes from 1 µm to 10 µm at a distance of 2.5 m. At this operation range, the signal from single particles of all sizes was successfully acquired enabling particle velocity measurements through the Doppler shifted frequency along the beam axis. For the particular case of 10 µm particles, single aerosol particles were still detected at an unprecedented range of 10 m. A theoretical treatment describing the relation between Mie scattering theory and the self-mixing phenomenon on single-particle detection is presented supporting the experimental results. The results show that the edge-filter enhanced self-mixing technique opens new possibilities for self-mixing detection where longer ranges, lower backscattering laser powers and higher velocities are involved. For example, it can be used as a robust and inexpensive anemometer for LDV applications for airflows with low-number density of microparticles.

Edge filter enhanced self-mixing interferometry.

Self-mixing interferometry (SMI) represents a simple, robust, and self-aligned technique for metrology applications. Still, its applicability on diffuse targets is limited to distances up to few meters. We present an enhanced approach based on the detection of the frequency-modulated (FM) self-mixing signal. The FM signal detection is achieved using an acetylene edge filter that maps laser frequency variations into intensity variations as the laser wavelength is tuned to the edge of the steep absorption profile. An experimental comparison between the enhanced and the conventional SMI approach is presented. The new approach yields to about two orders of magnitude larger signal-to-noise ratio and extends the applicability of SMI into new fields allowing longer detection ranges and lower backscattering signals.

Enhanced standing-wave acoustic levitation using high-order transverse modes in phased array ultrasonic cavities.

Airborne acoustic trapping by ultrasonic phased arrays has seen great advances in recent years, and yet the manipulation of objects with different shapes and sizes or heavy particles remains challenging. Here, we demonstrate that the manipulation capabilities of a standing-wave acoustic levitator can be extended by introducing intracavity high-order transverse (HOT) modes in the azimuthal direction, enabling the simultaneous trapping of several objects within a wide range of shapes and sizes with positional and rotational stability, including objects with sizes larger than one wavelength and weights in the scale of millinewtons. The conditions to generate different HOT modes are theoretically analyzed and experimentally implemented. We numerically calculate the pressure distributions, exhibiting good qualitative agreement with the experimental pressure distributions obtained with schlieren images. In addition, we calculate the acoustic force field for several examples of HOT modes and different particle sizes, which leads to a qualitative understanding of the experimental observations.

The resonant behavior of airborne standing-wave acoustic levitators based on arrays of ultrasonic transducers.

Recently airborne standing-wave acoustic levitation has seen great advances, and its applicability has been broadened due to the development of cavities constructed with arrays of compact ultrasonic sources. Yet, the numerical methods employed to study and predict the pressure distributions inside these cavities do not consider the effect of multiple reflections on the boundaries, hiding their resonant effects. This work presents an analytical, numerical, and experimental study of the effect of multiple reflections inside ultrasonic cavities based on arrays of transducers exhibiting their influence on the pressure amplitudes of focused standing waves. Our numerical results come from a modified version of the Matrix Method to numerically compute the multiple wave reflections of cavities constructed by two opposite arrays of multiple compact sources as boundaries. The correlation between numerical and experimental results reveals that intra-cavity reflections are relevant in focused axisymmetric cavities based on two arrays of multiple ultrasonic sources having a considerable impact on the amplitude of the standing waves and consequently, on the acoustic levitation performance. Thus, intra-cavity reflections must be considered for optimal cavity designs.

Modifying the Ambient Light Spectrum Using LED Lamps Alters the Phenolic Profile of Hydroponically Grown Greenhouse Lettuce Plants without Affecting Their Agronomic Characteristics.

The growth and development of green lettuce plants can be modulated by the prevailing light conditions around them. The aim of this study was to evaluate the effect of ambient light enrichment with different LED light spectra on agronomic characteristics, polyphenol concentration and relative gene expression of enzymes associated with polyphenol formation in 'Levistro' lettuce grown hydroponically in a Nutrient Film Technique (NFT) system for 28 days in a greenhouse. The spectra (blue:green:red:far-red) and red:blue (R:B) ratios obtained by enriching ambient light with Blue (B), White (W), Blue-Red (BR) and Red (R) LED light were B: 47:22:21:10, 0.5:1; W: 30:38:23:9, 0.8:1; BR: 33:15:44:8, 1.3:1 and R: 16:16:60:8, 3.8:1, respectively, and photosynthetically active radiation (PAR) under the different treatments, measured at midday, ranged from 328 to 336 µmoles m-2 s-1. The resulting daily light integral (DLI) was between 9.1 and 9.6 mol m-2 day-1. The photoperiod for all enrichment treatments was 12 h of light. The control was ambient greenhouse light (25:30:30:15; R:B = 1.2:1; PAR = 702 µmoles m-2 s-1; DLI = 16.9 mol m-2 day-1; photoperiod = 14.2 h of light). Fresh weight (FW) and dried weight percentage (DWP) were similar among the enrichment treatments and the control. The leaf number increased significantly under BR and R compared to B lights. The relative index of chlorophyll concentration (RIC) increased as plants grew and was similar among the enrichment treatments and the control. On the other hand, the concentration of chlorogenic acid and chicoric acid increased under BR and B lights, which was consistent with the higher relative expression of the coumarate 3-hydroxylase enzyme gene. In view of the results, it is inferred that half of the PAR or DLI is sufficient to achieve normal growth and development of 'Levistro' lettuce plants, suggesting a more efficient use of light energy under the light enrichment treatments. On the other hand, the blue and combined blue-red lights promoted the accumulation of phenolic compounds in the leaves of 'Levistro' lettuce plants.

Simulation-Based Training Program for Peripherally Inserted Central Catheter Placement: Randomized Comparative Study of in-Person Training With Synchronous Feedback Versus Distance Training With Asynchronous Feedback.

INTRODUCTION: Simulation training that includes deliberate practice is effective for procedural skill training. Delivering feedback remotely and asynchronously has been examined for more cost-efficient training. This prospective randomized study aimed to compare 2 feedback techniques for simulation training: synchronous direct feedback versus asynchronous distance feedback (ASYNC). METHODS: Forty anesthesia and internal medicine residents were recruited after study approval by the institutional ethics committee. Residents reviewed instructional material on an online platform and performed a pretraining assessment (PRE) for peripherally inserted central catheter (PICC) placement. Each resident was then randomly assigned to 1 of 2 training types, practice with synchronous direct feedback (SYNC) or practice with ASYNC. Training consisted of four, 1-hour practice sessions; each was conducted once per week. Both groups underwent posttraining evaluation (POST). The PRE and POST assessments were videotaped and evaluated by 2 independent, blinded reviewers using a global rating scale. RESULTS: Thirty-five residents completed the training program and both evaluations. Both groups had significantly improved global rating scale scores after 4 sessions. The SYNC group improved from 28 to 45 points (P < 0.01); the ASYNC group improved from 26.5 to 46 points (P < 0.01). We found no significant between-group differences for the PRE (P = 0.42) or POST assessments (P = 0.13). CONCLUSION: This simulation-based training program significantly improved residents' peripherally inserted central venous catheter placement skills using either modality. With these results, we are unable to demonstrate the superiority of synchronous feedback over ASYNC. Asynchronous feedback training modality represents a new, innovative approach for health care procedural skills training.

Oral infectivity through carnivorism in murine model of Trypanosoma cruzi infection.

Introduction: Oral transmission of T. cruzi is probably the most frequent transmission mechanism in wild animals. This observation led to the hypothesis that consuming raw or undercooked meat from animals infected with T. cruzi may be responsible for transmitting the infection. Therefore, the general objective of this study was to investigate host-pathogen interactions between the parasite and gastric mucosa and the role of meat consumption from infected animals in the oral transmission of T. cruzi. Methods: Cell infectivity assays were performed on AGS cells in the presence or absence of mucin, and the roles of pepsin and acidic pH were determined. Moreover, groups of five female Balb/c mice were fed with muscle tissue obtained from mice in the acute phase of infection by the clone H510 C8C3hvir of T. cruzi, and the infection of the fed mice was monitored by a parasitemia curve. Similarly, we assessed the infective capacity of T. cruzi trypomastigotes and amastigotes by infecting groups of five mice Balb/c females, which were infected orally using a nasogastric probe, and the infection was monitored by a parasitemia curve. Finally, different trypomastigote and amastigote inoculums were used to determine their infective capacities. Adhesion assays of T. cruzi proteins to AGS stomach cells were performed, and the adhered proteins were detected by western blotting using monoclonal or polyclonal antibodies and by LC-MS/MS and bioinformatics analysis. Results: Trypomastigote migration in the presence of mucin was reduced by approximately 30%, whereas in the presence of mucin and pepsin at pH 3.5, only a small proportion of parasites were able to migrate (∼6%). Similarly, the ability of TCTs to infect AGS cells in the presence of mucin is reduced by approximately 20%. In all cases, 60-100% of the animals were fed meat from mice infected in the acute phase or infected with trypomastigotes or amastigotes developed high parasitemia, and 80% died around day 40 post-infection. The adhesion assay showed that cruzipain is a molecule of trypomastigotes and amastigotes that binds to AGS cells. LC-MS/MS and bioinformatics analysis, also confirmed that transialidase, cysteine proteinases, and gp63 may be involved in TCTs attachment or invasion of human stomach cells because they can potentially interact with different proteins in the human stomach mucosa. In addition, several human gastric mucins have cysteine protease cleavage sites. Discussion: Then, under our experimental conditions, consuming meat from infected animals in the acute phase allows the T. cruzi infection. Similarly, trypomastigotes and amastigotes could infect mice when administered orally, whereas cysteinyl proteinases and trans-sialidase appear to be relevant molecules in this infective process.

Effectiveness of Teaching Mini Handball through Non-Linear Pedagogy in Different Socioeconomic Contexts: A Pilot Study.

Mini handball is among the sports included as part of school physical education in Chile to improve children's motor skills and to motivate their adherence to a healthy and active lifestyle in response to concerns about this country's high level of childhood obesity. To this end, non-linear pedagogy (NLP) has been used to develop motor skills through mini handball in the school context. However, socioeconomic differences that influence the development of children's motor skills have not been considered to determine whether the methodology applies to everyone. The aim of the present observational study is to describe and compare the effectiveness of the previously applied NLP methodology in two contrasting socioeconomic contexts to determine whether it helps to develop motor skills through mini handball in both school contexts. The Levine test was used to determine the homogeneity of the variances (p < 0.05), as the distribution of the data was not normal. The Kruskal-Wallis H statistical test was used to analyse within-group data. Additionally, the Mann-Whitney U test was applied for comparisons between groups. The results show significant improvements in the acquisition of the expected motor skills specific to mini handball. Additionally, a shortening of the gap was evidenced between the groups during the training process, with no significant differences at the end of the progression. Therefore, the investigated NLP is equally as effective for schoolchildren in two opposite socioeconomic contexts.

Adjusting single-axis acoustic levitators in real time using rainbow schlieren deflectometry.

Acoustic levitation uses focused high-intensity airborne ultrasound to hold particles in mid-air. It is becoming an important tool for experiments in spectrometry, lab-on-a-droplet, and display technologies. Nowadays, arrays of multiple small transducers can be used to build acoustic levitators; however, their performance depends on the optimal alignment. This work describes a simple method capable of visualizing a 2D projection of the acoustic field in real time using rainbow schlieren deflectometry. Good agreement was found between the images obtained with this technique and simulations of the acoustic pressure. It was also found that the maximum amplitudes of the field were obtained with the levitator aligned so that the power consumption was minimum, showing another simple and affordable way to adjust the levitators. As a result of the alignment optimization, it was possible for the first time to levitate steel and mercury in a levitator constructed with off-the-shelf components. The schlieren technique was applied to the TinyLev acoustic levitation system, but it can be applied to visualize the acoustic potential produced by different types of levitation systems.

Characterization of pectin extracted under mild conditions from tempesquistle (Sideroxylon palmeri) fruit at two maturity stages.

BACKGROUND: le tree (Sideroxylon palmeri) belongs to family Sapotaceae, and its fruits contain hydrophilic and hydrophobic gums. Aim of this study was to develop an extraction method for pectin, a hydrophilic gum, from green and ripe tempesquistle fruit. The extraction method was gentle to minimize potential structural changes in pectin, a maceration was used, and then pectin precipitation was induced with acetone at room temperature. Pectin characterization was carried out in two steps. The first step consisted of qualitative tests of Molisch, Fehling and Lugol. The second step consisted of quantitative tests to determine esterification degree (ED) by ATR-FTIR and D-galacturonic acid (D-GalA) content with a colorimetric assay. ATR-FTIR spectroscopic method revealed that green and ripe tempesquistle fruit have an ED of 0% and 30.4%, respectively. Both fruit have a low ED pectin. The content of D-GalA in green and ripe pectins was 18.8 ±2.7% and 20.2 ±0.6%, respectively. The yield obtained in green and ripe fruit samples was 1.6 ±0.2% and 3.0 ±0.1%, respectively. The extraction method allowed two types of pectins obtained in function of maturity stage of tempesquistle fruit distinguishable by. METHODS: istle tree (Sideroxylon palmeri) belongs to family Sapotaceae, and its fruits contain hydrophilic and hydrophobic gums. Aim of this study was to develop an extraction method for pectin, a hydrophilic gum, from green and ripe tempesquistle fruit. The extraction method was gentle to minimize potential structural changes in pectin, a maceration was used, and then pectin precipitation was induced with acetone at room temperature. Pectin characterization was carried out in two steps. The first step consisted of qualitative tests of Molisch, Fehling and Lugol. The second step consisted of quantitative tests to determine esterification degree (ED) by ATR-FTIR and D-galacturonic acid (D-GalA) content with a colorimetric assay. ATR-FTIR spectroscopic method revealed that green and ripe tempesquistle fruit have an ED of 0% and 30.4%, respectively. Both fruit have a low ED pectin. The content of D-GalA in green and ripe pectins was 18.8 ±2.7% and 20.2 ±0.6%, respectively. The yield obtained in green and ripe fruit samples was 1.6 ±0.2% and 3.0 ±0.1%, respectively. The extraction method allowed two types of pectins obtained in function of maturity stage of tempesquistle fruit distinguishable by ATR. RESULTS: istle tree (Sideroxylon palmeri) belongs to family Sapotaceae, and its fruits contain hydrophilic and hydrophobic gums. Aim of this study was to develop an extraction method for pectin, a hydrophilic gum, from green and ripe tempesquistle fruit. The extraction method was gentle to minimize potential structural changes in pectin, a maceration was used, and then pectin precipitation was induced with acetone at room temperature. Pectin characterization was carried out in two steps. The first step consisted of qualitative tests of Molisch, Fehling and Lugol. The second step consisted of quantitative tests to determine esterification degree (ED) by ATR-FTIR and D-galacturonic acid (D-GalA) content with a colorimetric assay. ATR-FTIR spectroscopic method revealed that green and ripe tempesquistle fruit have an ED of 0% and 30.4%, respectively. Both fruit have a low ED pectin. The content of D-GalA in green and ripe pectins was 18.8 ±2.7% and 20.2 ±0.6%, respectively. The yield obtained in green and ripe fruit samples was 1.6 ±0.2% and 3.0 ±0.1%, respectively. The extraction method allowed two types of pectins obtained in function of maturity stage of tempesquistle fruit distinguishable by ATR-FTIR.