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We used light-sensitive drugs to identify the brain region-specific role of mGlu5 metabotropic glutamate receptors in the control of pain. Optical activation of systemic JF-NP-26, a caged, normally inactive, negative allosteric modulator (NAM) of mGlu5 receptors, in cingulate, prelimbic, and infralimbic cortices and thalamus inhibited neuropathic pain hypersensitivity. Systemic treatment of alloswitch-1, an intrinsically active mGlu5 receptor NAM, caused analgesia, and the effect was reversed by light-induced drug inactivation in the prelimbic and infralimbic cortices, and thalamus. This demonstrates that mGlu5 receptor blockade in the medial prefrontal cortex and thalamus is both sufficient and necessary for the analgesic activity of mGlu5 receptor antagonists. Surprisingly, when the light was delivered in the basolateral amygdala, local activation of systemic JF-NP-26 reduced pain thresholds, whereas inactivation of alloswitch-1 enhanced analgesia. Electrophysiological analysis showed that alloswitch-1 increased excitatory synaptic responses in prelimbic pyramidal neurons evoked by stimulation of presumed BLA input, and decreased BLA-driven feedforward inhibition of amygdala output neurons. Both effects were reversed by optical silencing and reinstated by optical reactivation of alloswitch-1. These findings demonstrate for the first time that the action of mGlu5 receptors in the pain neuraxis is not homogenous, and suggest that blockade of mGlu5 receptors in the BLA may limit the overall analgesic activity of mGlu5 receptor antagonists. This could explain the suboptimal effect of mGlu5 NAMs on pain in human studies and validate photopharmacology as an important tool to determine ideal target sites for systemic drugs.

Summary: Metabotropic glutamate receptors (mGlu) regulate multiple functions in the nervous systems and are involved in several neurological disorders. However, selectively targeting individual mGlu subtypes with spatiotemporal precision is still an unmet need. Photopharmacology can address this concern through the utilization of photoswitchable compounds such as optogluram, which is a positive allosteric modulator (PAM) of mGlu4 that enables the precise control of physiological responses using light but does not have an optimal selectivity profile. Optogluram analogs were developed to obtain photoswitchable PAMs of mGlu4 receptor with an improved selectivity. Among them, optogluram-2 emerged as a photoswitchable ligand for mGlu4 receptor with activity as both PAM and allosteric agonists. It presents a higher selectivity and offers improved photoswitching of mGlu4 activity. These improved properties make optogluram-2 an excellent candidate to study the role of mGlu4 with a high spatiotemporal precision in systems where mGlu4 can be co-expressed with other mGlu receptors.

The high recurrence rate of cystine lithiasis observed in cystinuria patients highlights the need for new therapeutic options to address this chronic disease. There is growing evidence of an antioxidant defect in cystinuria, which has led to test antioxidant molecules as new therapeutic approaches. In this study, the antioxidant l-Ergothioneine was evaluated, at two different doses, as a preventive and long-term treatment for cystinuria in the Slc7a9−/− mouse model. l-Ergothioneine treatments decreased the rate of stone formation by more than 60% and delayed its onset in those mice that still developed calculi. Although there were no differences in metabolic parameters or urinary cystine concentration between control and treated mice, cystine solubility was increased by 50% in the urines of treated mice. We also demonstrate that l-Ergothioneine needs to be internalized by its transporter OCTN1 (Slc22a4) to be effective, as when administrated to the double mutant Slc7a9−/−Slc22a4−/− mouse model, no effect on the lithiasis phenotype was observed. In kidneys, we detected a decrease in GSH levels and an impairment of maximal mitochondrial respiratory capacity in cystinuric mice that l-Ergothioneine treatment was able to restore. Thus, l-Ergothioneine administration prevented cystine lithiasis in the Slc7a9−/− mouse model by increasing urinary cystine solubility and recovered renal GSH metabolism and mitochondrial function. These results support the need for clinical trials to test l-Ergothioneine as a new treatment for cystinuria.

The physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast, and reversible control of endogenous protein activity, with spatiotemporal resolution set by the illumination method. Here, we combine a photoswitchable allosteric modulator (alloswitch) and 2-photon excitation using pulsed near-infrared lasers to reversibly silence metabotropic glutamate 5 (mGlu5) receptor activity in intact brain tissue. Endogenous receptors can be photoactivated in neurons and astrocytes with pharmacological selectivity and with an axial resolution between 5 and 10 µm. Thus, 2-photon pharmacology using alloswitch allows investigating mGlu5-dependent processes in wild-type animals, including synaptic formation and plasticity, and signaling pathways from intracellular organelles.

Purpose: Script training is a well-established treatment for aphasia, but its evidence comes almost exclusively from monolingual English speakers with aphasia. Furthermore, its active ingredients and profiles of people with aphasia (PWA) that respond to this treatment remain understudied. This study aimed to adapt a scripted-sentence learning protocol to Colombian Spanish speakers with aphasia, investigate speech entrainment (i.e., unison production of sentences) as an active ingredient for scripted-sentence learning, and identify patient profiles associated with better scripted-sentence learning. Method: Fourteen monolingual Spanish speakers with aphasia learned a set of 30 sentences. To examine speech entrainment as an active ingredient for scripted-sentence learning, we investigated whether sentences containing externally added rhythmic cues (involving stress-aligned vs. metronomic rhythmic cues) would result in better scripted-sentence learning compared with control sentences. Learning was measured via postsession probes and analyzed using mixed-effects logistic regression models. The relationship between scripted-sentence learning and baseline language and rhythmic processing measures was also examined. Results: Significant scripted-sentence learning over time indicated a successful adaptation of a script-training protocol to Spanish. PWA learned significantly more scripted sentences in the rhythmically enhanced conditions compared with the control condition. There were no differences between rhythmically enhanced conditions (stress-aligned vs. metronomic). In terms of patient profiles, it was found that PWA with more severe aphasia demonstrated larger learning gains, but rhythmic processing showed little association with learning estimates. Conclusions: To our knowledge, this study provides the first adaptation of a scripted-sentence learning protocol for monolingual Spanish speakers with aphasia, demonstrating cross-linguistic benefits of script training interventions. Highlighting rhythmic features during speech entrainment facilitated scripted-sentence learning in Spanish speakers with aphasia, suggesting that speech entrainment may be an active ingredient for scripted-sentence learning. More severe aphasia was associated with better scripted-sentence learning, suggesting that more severely impaired individuals are likely to benefit most from this treatment. Supplemental Material: https://doi.org/10.23641/asha.19241847, Understanding the connection between active ingredients and patient profiles associated with better response to evidence-based aphasia treatments is a necessary step to improve both the implementation and individualization of aphasia [...]

Summary: Adrenoceptors are G protein-coupled receptors involved in a large variety of physiological processes, also under pathological conditions. This is due in large part to their ubiquitous expression in the body exerting numerous essential functions. Therefore, the possibility to control their activity with high spatial and temporal precision would constitute a valuable research tool. In this study, we present a caged version of the approved non-selective β-adrenoceptor antagonist carvedilol, synthesized by alkylation of its secondary amine with a coumarin derivative. Introducing this photo-removable group abolished carvedilol physiological effects in cell cultures, mouse isolated perfused hearts and living zebrafish larvae. Only after visible light application, carvedilol was released and the different physiological systems were pharmacologically modulated in a similar manner as the control drug. This research provides a new photopharmacological tool for a wide range of research applications that may help in the development of future precise therapies.

Lepidopteran pests are major factors limiting soybean productivity in South America. In some cases, effective management of these species requires the use of foliar insecticides. For sustainable use of these insecticides, they should only be applied when insect population size exceeds an economic threshold. Since this estimation requires to determine the consumption of different species, this work aimed to integrate all these factors, studying the consumption of small (less than 1 cm long) and medium (1 to 1.5 cm long) size larvae of major lepidopteran pests to vegetative and reproductive tissues on Bt (M7739IPRO variety, containing the event MON87701 which expresses the Cry1Ac protein from Bacillus thuringiensis) and non-Bt (BMX Desafio RR variety) soybeans. The feeding injury to vegetative tissues was tested in detached-leaf assays in grow chambers, and for reproductive structures the study was conducted in greenhouse with infestations at early (flowering) and mid reproductive (mid grain filling) stages. Based on the feeding behavior of the species tested, they were cast in four groups: a) Anticarsia gemmatalis and Chrysodeixis includens, defoliating only the RR variety with the lowest consumption of foliar area; b) Spodoptera eridania, defoliating both RR and IPRO varieties, consuming twice than the species mentioned above; c) Helicoverpa armigera, defoliating and being the most damaging species to pods in the RR variety; and d) S. cosmioides and S. frugiperda, defoliating and damaging pods in both varieties. The species differed in their ability to feed on IPRO varieties, so a different economic threshold should be considered. Consequently, in cases where more than one species are found simultaneously, the species composition should be considered in estimating the economic threshold. Additionally, our findings may contribute to a better decision-making to control insect feeding injury in IPRO varieties, because a slower larval growth provides more time to ensure the need of control with insecticides. In summary, this clasification contributes to an improved recommendation of sustainable insecticide use, taking into account the behavior of each species that are major soybeans pests in South America.

In English, positions of lexical stress in disyllabic words are associated with word categories; that is, nouns tend to be stressed more often on the first syllable, whereas verbs are more likely to be stressed on the second syllable (i.e. "sub"ject (noun) vs. sub"ject" (verb)). This phenomenon, which is called the stress typicality effect, has been shown to facilitate word recognition to native English speakers. However, there is little research on whether it also facilitates word recognition to non-native speakers of English, in particular, to English learners with a tonal first language. To fill this research gap, the present study investigated whether the stress typicality effect modulated word recognition in native speakers of Chinese who learned English as a second language. Both visual grammatical classification and lexical decision tasks were administered to ESL learners with intermediate and advanced English proficiency. The results revealed that Chinese intermediate ESL learners were not sensitive to stress typicality in English; however, the advanced learners were. The findings suggest that different performances in stress assignment among Chinese ESL learners were influenced by their English proficiency levels.

Metabotropic glutamate 5 receptors (mGlu5) are thought to play an important role in mediating emotional information processing. In particular, negative allosteric modulators (NAMs) of mGlu5 have received a lot of attention as potential novel treatments for several neuropsychiatric diseases, including anxiety-related disorders. The aim of this study was to assess the influence of pre- and post-training mGlu5 inactivation in cued fear conditioned mice on neuronal oscillatory activity during fear retrieval. For this study we used the recently developed mGlu5 NAM Alloswicth-1 administered systemically. Injection of Alloswicth-1 before, but not after, fear conditioning resulted in a significant decrease in freezing upon fear retrieval. Mice injected with Alloswicth-1 pre-training were also implanted with recording microelectrodes into both the medial prefrontal cortex (mPFC) and ventral hippocampus (vHPC). The recordings revealed a reduction in theta rhythmic activity (4–12 Hz) in both the mPFC and vHPC during fear retrieval. These results indicate that inhibition of mGlu5 signaling alters local oscillatory activity in principal components of the fear brain network underlying a reduced response to a predicted threat.

Cystinuria, a rare inherited aminoaciduria condition, is characterized by the hyperexcretion of cystine, ornithine, lysine, and arginine. Its main clinical manifestation is cystine stone formation in the urinary tract, being responsible for 1–2% total and 6–8% pediatric lithiasis. Cystinuria patients suffer from recurrent lithiasic episodes that might end in surgical interventions, progressive renal functional deterioration, and kidney loss. Cystinuria is monitored for the presence of urinary cystine stones by crystalluria, imaging techniques or urinary cystine capacity; all with limited predicting capabilities. We analyzed blood and urine levels of the natural antioxidant L-ergothioneine in a Type B cystinuria mouse model, and urine levels of its metabolic product S-methyl-L-ergothioneine, in both male and female mice at two different ages and with different lithiasic phenotype. Urinary levels of S-methyl-L-ergothioneine showed differences related to age, gender and lithiasic phenotype. Once normalized by L-ergothioneine to account for interindividual differences, the S-methyl-L-ergothioneine to L-ergothioneine urinary ratio discriminated between cystine lithiasic phenotypes. Urine S-methyl-L-ergothioneine to L-ergothioneine ratio could be easily determined in urine and, as being capable of discriminating between cystine lithiasis phenotypes, it could be used as a lithiasis biomarker in cystinuria patient management.

This paper documents for the first time the intonation system of Tucumán Spanish, an understudied variety of Argentinian Spanish. Semi-spontaneous speech illustrating the intonation of main sentence types, i.e. broad focus statements, partial and absolute interrogatives, and imperatives and vocatives, was elicited from 31 native speakers of Tucumán Spanish via an adapted version of the Argentinian Intonation Survey (Prieto and Roseano, 2009-2013). The two authors listened to the recordings and transcribed them using the Tones and Break Indexes conventions (ToBI) (Beckman et al. 2002, Prieto and Roseano 2010, Hualde and Prieto 2015). Transcriptions of prenuclear and nuclear configurations together with their respective frequencies allowed both an appreciation of the most used configurations within each sentence type along with detailed variation at the phonetic level. For example, yes/no questions were consistently realized with a low nuclear pitch accent L* and an ascending boundary tone. However, there was variation in the height of the boundary tones yielding the frequent contour L* ¡H%, and the less frequent L* H%. Altogether, these detailed patterns document the systematic phonetic variation of the intonation system of TS and provide a basis for future research to determine the phonological status of this variation.

Resumen: Este trabajo describe el proceso de revisión del catálogo de servicios de salud pública de la ciudad de Barcelona para responder a la reducción presupuestaria producida a partir de 2010 en las administraciones públicas en España. Se basó en un grupo de trabajo que revisó las diferentes líneas de actividad, teniendo en cuenta los gastos y los ingresos asociados y su razón de ser, valorando factores como la obligación legal de desempeño, la financiación finalista, la petición explícita por las administraciones fundadoras y la existencia o no de otros actores capaces de asumirla. Se describen los cambios y sus consecuencias. Se valora que el nuevo catálogo está consolidado y se considera ratificado por los órganos de gobierno, que se han renovado tras los relevos políticos. Creemos que esto es fruto de haberlo basado en el consenso profesional y los criterios de gestión, claves para el buen funcionamiento de un organismo autónomo ejecutivo de carácter público. Abstract: This paper describes the review process of the Agency of Public Health of Barcelona's service portfolio in response to the budget cuts introduced since 2010 in the public administrations in Spain. A working group reviewed the different business activities, taking into account their costs and generated revenue and their justification, assessing factors such as the existence of legal constraints, tied funding, explicit demands from the founding administrations and other actors that may be capable of undertaking particular activities. The changes and their consequences are described. The new service portfolio has been consolidated and is considered ratified by the Agency board, which was renewed after political changes. We conclude that this is because it was based on professional consensus and management criteria, which are key for the smooth operation of a public autonomous executive organisation. Palabras clave: Administración de salud pública, Organización y administración, Administración pública, Práctica de la salud pública, Keywords: Public health administration, Organisation and administration, Public administration, Public health practice

Biological activity can be photoswitched by light-regulated drugs, but so far only diffusible ligands have been shown to work on endogenous receptors. Here the authors develop targeted covalent photoswitches that couple to a protein target by ligand affinity, and demonstrate photocontrol of GluK1-expressing neurons.

Although there is consistent evidence that higher levels of processing, such as learning the form-meaning associations specific to the second language (L2), are a source of difficulty in acquiring L2 speech, no study has addressed how these levels interact in shaping L2 perception and production of intonation. We examine the hypothesis of whether access to contextual meaning increases the chances of first language (L1) influence on L2 intonation. To test this hypothesis, we compared the perception and production of sentential English focus by 27 advanced English language learners (n = 13 L1 Mandarin speakers; n = 14 L1 Spanish speakers) and 13 controls, through a series of tasks that promoted different levels of access to meaning. Results showed that L1 transfer was especially clear in Spanish speakers. Not only did they consistently differ from controls in their perception of focalized verbs and subjects, showing their L1 bias to perceive focus at the end of a sentence, but they were also the only group of speakers that inserted pauses after the focalized word, which showed strong L1 effects. Moreover, these L1 transfer effects were more obvious in contextualized tasks, which indicated that facilitating access to meaning by adding context increased L1 transfer effects on the perception and especially on the production of focus intonation.

Metabotropic glutamate receptors (mGlus) are G Protein coupled-receptors that modulate synaptic transmission and plasticity in the central nervous system. Some act as autoreceptors to control neurotransmitter release at excitatory synapses and have become attractive targets for drug therapy to treat certain neurological disorders. However, the high degree of sequence conservation around the glutamate binding site makes the development of subtype-specific orthosteric ligands difficult to achieve. This problem can be circumvented by designing molecules that target specific less well conserved allosteric sites. One such allosteric drug, the photo-switchable compound OptoGluNAM4.1, has been recently employed to reversibly inhibit the activity of metabotropic glutamate 4 (mGlu4) receptors in cell cultures and in vivo. We studied OptoGluNAM4.1 as a negative modulator of neurotransmission in rodent cerebellar slices at the parallel fiber – Purkinje cell synapse. Our data show that OptoGluNAM4.1 antagonizes pharmacological activation of mGlu4 receptors in a fully reversible and photo-controllable manner. In addition, for the first time, this new allosteric modulator allowed us to demonstrate that, in brain slices from the rodent cerebellar cortex, mGlu4 receptors are endogenously activated in excitotoxic conditions, such as the early phases of simulated cerebellar ischemia, which is associated with elevated levels of extracellular glutamate. These findings support OptoGluNAM4.1 as a promising new tool for unraveling the role of mGlu4 receptors in the central nervous system in physio-pathological conditions.

Experimental animal models are unique tools (i) to study pain transmission and pathophysiology of neuropathic pain, (ii) to identify novel molecular targets and (iii) to test the potential analgesic effect of specific molecules. The chronic constriction injury (CCI) model of neuropathic pain is the first model of post-traumatic painful peripheral neuropathy, originally developed by Bennett and Xie in the late 1980s. The chronic constriction is performed in the sciatic nerve and induces a partial denervation involving myelinated afferent axons and unmyelinated axons. Damage to unmyelinated axons is much more severe than myelinated afferents. As the model induces a partial denervation, it is very useful for the analysis of pain behaviours. Stimulation of the hind paw, a target of the sciatic nerve, induces pain which can be quantitated. Thus, mechanical allodynia is usually assessed 7, 14 and 21 days after CCI of the sciatic nerve by measuring the hind paw withdrawal response to von Frey filament stimulation. Here, we describe in detail the protocol allowing a reliable and reproducible CCI model in mice. Overall, researchers most commonly use this surgical model to discover more efficacious drugs for the pharmacological control of chronic pain states.

Pain research is mostly based on experimental assays that use animal models, which may allow deciphering the physiopathology of this condition and to propel drug discovery. The formalin nociception test is considered one of the most predictive approaches to study acute pain in rodents. This test permits monitoring pain-related responses (i.e., itch) caused by a subcutaneous injection of an inflammatory agent, namely 2.5% formalin solution, in the hind paw. After the injection, two distinct time periods or phases of licking/biting behaviour occur, which are separated by a quiescent period. Importantly, these phases differ in duration and underlying mechanisms. Hence, the initial acute phase (phase I), commonly recorded for 5 min just after formalin administration, reflects acute peripheral pain, probably due to direct activation of nociceptors through TRPA1 channels. On the other hand, the phase II, which starts after the quiescent period (5-15 min) and is commonly recorded for 15-30 min, is due to the ongoing inflammatory input and central nociceptive sensitization. Here, we describe in detail the protocol used to perform a reliable and reproducible formalin test in mice.

GPCRs play critical roles in cell communication. Although GPCRs can form heteromers, their role in signaling remains elusive. Here we used rat metabotropic glutamate (mGlu) receptors as prototypical dimers to study the functional interaction between each subunit. mGluRs can form both constitutive homo- and heterodimers. Whereas both mGlu2 and mGlu4 couple to G proteins, G protein activation is mediated by mGlu4 heptahelical domain (HD) exclusively in mGlu2-4 heterodimers. Such asymmetric transduction results from the action of both the dimeric extracellular domain, and an allosteric activation by the partially-activated non-functional mGlu2 HD. G proteins activation by mGlu2 HD occurs if either the mGlu2 HD is occupied by a positive allosteric modulator or if mGlu4 HD is inhibited by a negative modulator. These data revealed an oriented asymmetry in mGlu heterodimers that can be controlled with allosteric modulators. They provide new insight on the allosteric interaction between subunits in a GPCR dimer.

Light-operated drugs constitute a major target in drug discovery, since they may provide spatiotemporal resolution for the treatment of complex diseases (i.e. chronic pain). JF-NP-26 is an inactive photocaged derivative of the metabotropic glutamate type 5 (mGlu5) receptor negative allosteric modulator raseglurant. Violet light illumination of JF-NP-26 induces a photochemical reaction prompting the active-drug’s release, which effectively controls mGlu5 receptor activity both in ectopic expressing systems and in striatal primary neurons. Systemic administration in mice followed by local light-emitting diode (LED)-based illumination, either of the thalamus or the peripheral tissues, induced JF-NP-26-mediated light-dependent analgesia both in neuropathic and in acute/tonic inflammatory pain models. These data offer the first example of optical control of analgesia in vivo using a photocaged mGlu5 receptor negative allosteric modulator. This approach shows potential for precisely targeting, in time and space, endogenous receptors, which may allow a better management of difficult-to-treat disorders.

The general literature on the phonetic correlates of stress agrees that duration, and in stress accent languages, are consistent correlates of stress. However, the role of amplitude changes in the speech signal is more controversial. In particular, the conflicting results of spectral tilt as a correlate of stress have been attributed to the effects of vowel reduction. We examined the stress correlates of duration, overall intensity and spectral tilt in Catalan and Spanish in both accented and unaccented contexts while controlling for formant frequency differences between morphologically corresponding vowels in stressed and unstressed environments by comparing vowels that maintain the same quality across stress contexts with those that do not. Duration was a consistent stress correlate in all vowels in both languages, regardless of their formant frequency differences across stress contexts and of the absence of pitch accents. In fact, stress-related formant frequency differences between corresponding vowels amplify the duration cues to the stress contrast. On the other hand, the use speakers made of intensity was not as pervasive as that of duration. Specifically, changes in spectral tilt were significant only in Catalan and in those vowels that alternate a more open and peripheral realization in stressed syllables with a mid-central realization in unstressed syllables, indicating that spectral tilt is related to the formant frequency differences linked to the centralization processes rather than to the stress contrast. (Contains 8 figures, 3 tables and 4 notes.)