Phenolic compound abundance in Pak choi leaves is controlled by salinity and dependent on pH of the leaf apoplast

Onset of salinity induces the pH of the leaf apoplast of Pak choi transiently to increase over a period of 2 to 3 hr. This pH event causes protein abundances in leaves to increase. Among them are enzymes that are key for the phenylpropanoid pathway. To answer the questions whether this short-term salt stress also influences contents of the underlying phenylpropanoids and for clarifying as to whether the apoplastic pH transient plays a role for such a putative effect, Pak choi plants were treated with 37.5 mM CaCl2 against a non-stressed control. A third experimental group, where the leaf apoplast of plants treated with 37.5 mM CaCl2, was clamped in the acidic range by means of infiltration of 5 mM citric acid/sodium citrate (pH 3.6), enabled validation of pH-dependent effects. Microscopy-based live cell imaging was used to quantify leaf apoplastic pH in planta. Phenolics were quantified shortly after the formation of the leaf apoplastic pH transient by means of HPLC-DAD-ESI-MS. Results showed that different phenolic compounds were modulated at 150 and 200 min after the onset of chloride salinity. A pH-independent reduction in phenolic acid abundance as well as an accumulation of phenolic acid:malate conjugates was quantified after 200 min of salt stress. However, at 150 min after the onset of salt stress, flavonoids were significantly reduced by salinity in a pH-dependent manner. These results provided a strong indication that the pH of the apoplast is a relevant component for the short-term metabolic response to chloride salinity.BMBF research grantDeutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/501100001659Peer Reviewe

The IMS Toucan System for the Blizzard Challenge 2023

For our contribution to the Blizzard Challenge 2023, we improved on the system we submitted to the Blizzard Challenge 2021. Our approach entails a rule-based text-to-phoneme processing system that includes rule-based disambiguation of homographs in the French language. It then transforms the phonemes to spectrograms as intermediate representations using a fast and efficient non-autoregressive synthesis architecture based on Conformer and Glow. A GAN based neural vocoder that combines recent state-of-the-art approaches converts the spectrogram to the final wave. We carefully designed the data processing, training, and inference procedures for the challenge data. Our system identifier is G. Open source code and demo are available.Comment: Published at the Blizzard Challenge Workshop 2023, colocated with the Speech Synthesis Workshop 2023, a sattelite event of the Interspeech 202

Quantitative analysis of muscle and tendon retraction in chronic rotator cuff tears

BACKGROUND: Musculotendinous retraction is a limiting factor for repair of long-standing rotator cuff tears. However, it is currently unknown to what extent the muscle and tendon contribute to the degree of total retraction. Further understanding of this may possibly influence the strategy of musculotendinous reconstruction. PURPOSE: To analyze the contribution of muscle and tendon to the process of myotendinous retraction. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: Magnetic resonance imaging of 130 shoulders with intact (n = 20) or completely torn supraspinatus tendons was analyzed. Fatty infiltration of the supraspinatus muscle was graded according to Goutallier stages. The degree of retraction of the tendon stump and of the musculotendinous junction was assessed. RESULTS: There were 30 shoulders without evidence of supraspinatus fatty infiltration, 25 with stage 1, 23 with stage 2, 25 with stage 3, and 15 with stage 4 changes. The corresponding tear sizes (distance of tendon end from greater tuberosity) were 4, 21, 27, 37, and 41 mm; the distance of the myotendinous junction from the greater tuberosity was 22, 33, 39, 48, and 48 mm; and the length of the tendons (distance of tendon end to myotendinous junction) was 19, 13, 12, 11, and 8 mm, respectively. In Goutallier stage 3 and above, and in case of a positive tangent sign, the musculotendinous junction was, in 90% of the cases, retracted to or beyond the glenoid. CONCLUSION: Musculotendinous retraction in chronic rotator cuff tears results mainly from shortening of the muscle fibers but in advanced stages results also from shortening of the tendon tissue itself. The present data demonstrate, for the first time, that the residual tendon stump in a tendon tear does not have the length of the original tendon and is further shortened over time. Therefore, direct anatomic tendon reinsertion will result in lengthening of the supraspinatus muscle greater than what it would have been before the tear

Comparison of in vitro and in vivo models for the elucidation of metabolic patterns of 7-azaindole-derived synthetic cannabinoids exemplified using cumyl-5F-P7AICA

Due to the dynamic market involving synthetic cannabinoids (SCs), the determination of analytical targets is challenging in clinical and forensic toxicology. SCs usually undergo extensive metabolism, and therefore their main metabolites must be identified for the detection in biological matrices, particularly in urine. Controlled human studies are usually not possible for ethical reasons; thus, alternative models must be used. The aim of this work was to predict the in vitro and in vivo metabolic patterns of 7‐azaindole‐derived SCs using 1‐(5‐fluoropentyl)‐N‐(2‐phenylpropan‐2‐yl)‐1H‐pyrollo[2,3‐b]pyridin‐3‐carboxamide (cumyl‐5F‐P7AICA) as an example. Different in vitro (pooled human liver S9 fraction, pooled human liver microsomes, and pig liver microsomes) and in vivo (rat and pig) systems were compared. Monooxygenase isoenzymes responsible for the most abundant phase I steps, namely oxidative defluorination (OF) followed by carboxylation, monohydroxylation, and ketone formation, were identified. In both in vivo models, OF/carboxylation and N‐dealkylation/monohydroxylation/sulfation could be detected. Regarding pHS9 and pig urine, monohydroxylation/sulfation or glucuronidation was also abundant. Furthermore, the parent compound could still be detected in all models. Initial monooxygenase activity screening revealed the involvement of CYP2C19, CYP3A4, and CYP3A5. Therefore, in addition to the parent compound, the OF/carboxylated and monohydroxylated (and sulfated or glucuronidated) metabolites can be recommended as urinary targets. In comparison to literature, the pig model predicts best the human metabolic pattern of cumyl‐5F‐P7AICA. Furthermore, the pig model should be suitable to mirror the time‐dependent excretion pattern of parent compounds and metabolites

Toxicokinetics of U-47700, tramadol, and their main metabolites in pigs following intravenous administration: is a multiple species allometric scaling approach useful for the extrapolation of toxicokinetic parameters to humans?

New synthetic opioids (NSOs) pose a public health concern since their emergence on the illicit drug market and are gaining increasing importance in forensic toxicology. Like many other new psychoactive substances, NSOs are consumed without any preclinical safety data or any knowledge on toxicokinetic (TK) data. Due to ethical reasons, controlled human TK studies cannot be performed for the assessment of these relevant data. As an alternative animal experimental approach, six pigs per drug received a single intravenous dose of 100 µg/kg body weight (BW) of U-47700 or 1000 µg/kg BW of tramadol to evaluate whether this species is suitable to assess the TK of NSOs. The drugs were determined in serum and whole blood using a fully validated method based on solid-phase extraction and LC–MS/MS. The concentration–time profiles and a population (pop) TK analysis revealed that a three-compartment model best described the TK data of both opioids. Central volumes of distribution were 0.94 L/kg for U-47700 and 1.25 L/kg for tramadol and central (metabolic) clearances were estimated at 1.57 L/h/kg and 1.85 L/h/kg for U-47700 and tramadol, respectively. The final popTK model parameters for pigs were upscaled via allometric scaling techniques. In comparison to published human data, concentration–time profiles for tramadol could successfully be predicted with single species allometric scaling. Furthermore, possible profiles for U-47700 in humans were simulated. The findings of this study indicate that unlike a multiple species scaling approach, pigs in conjunction with TK modeling are a suitable tool for the assessment of TK data of NSOs and the prediction of human TK data

The Antituberculosis Drug Ethambutol Selectively Blocks Apical Growth in CMN Group Bacteria

Members of the genus Mycobacterium are the most prevalent cause of infectious diseases. Mycobacteria have a complex cell envelope containing a peptidoglycan layer and an additional arabinogalactan polymer to which a mycolic acid bilayer is linked;this complex, multilayered cell wall composition (mAGP) is conserved among all CMN group bacteria. The arabinogalactan and mycolic acid synthesis pathways constitute effective drug targets for tuberculosis treatment. Ethambutol (EMB), a classical antituberculosis drug, inhibits the synthesis of the arabinose polymer. Although EMB acts bacteriostatically, its underlying molecular mechanism remains unclear. Here, we used Corynebacterium glutamicum and Mycobacterium phlei as model organisms to study the effects of EMB at the single-cell level. Our results demonstrate that EMB specifically blocks apical cell wall synthesis, but not cell division, explaining the bacteriostatic effect of EMB. Furthermore, the data suggest that members of the family Corynebacterineae have two dedicated machineries for cell elongation (elongasome) and cytokinesis (divisome). IMPORTANCE Antibiotic treatment of bacterial pathogens has contributed enormously to the increase in human health. Despite the apparent importance of antibiotic treatment of bacterial infections, surprisingly little is known about the molecular functions of antibiotic actions in the bacterial cell. Here, we analyzed the molecular effects of ethambutol, a first-line antibiotic against infections caused by members of the genus Mycobacterium. We find that this drug selectively blocks apical cell growth but still allows for effective cytokinesis. As a consequence, cells survive ethambutol treatment and adopt a pneumococcal cell growth mode with cell wall synthesis only at the site of cell division. However, combined treatment of ethambutol and beta-lactam antibiotics acts synergistically and effectively stops cell proliferation

Impact evaluation methods in public economics : a brief introduction to randomized evaluations and comparison with other methods

Recent years have seen a large expansion in the use of rigorous impact evaluation techniques. Increasingly, public administrations are collaborating with academic economists and other quantitative social scientists to apply such rigorous methods to the study of public finance. These developments allow for more reliable measurements of the effects of different policy options on the behavioral responses of citizens, firm owners, or public officials. They can help decision makers in tax administrations, public procurement offices, and other public agencies design programs informed by well-founded evidence. This article provides an introductory overview of the most frequently used impact evaluation methods. It is aimed at facilitating communication and collaboration between practitioners and academics by introducing key vocabulary and concepts used in rigorous impact evaluation methods, starting with randomized controlled trials and comparing them with other methods ranging from simple pre–post analysis to difference-in-differences, matching estimations, and regression discontinuity designs