Activation of cholinergic basal forebrain neurons improved cognitive functions in adult-onset hypothyroid mice.

Cognitive dysfunction is common in hypothyroid patients, even after undergoing sufficient levothyroxine (LT4) replacement therapy for euthyroid. Our previous studies indicated that cholinergic neurons might contribute to the decline of cognition in adult-onset hypothyroidism. Nevertheless, the role of the cellular and neural control of basal forebrain (BF) cholinergic neurons in hypothyroidism-induced cognitive impairments is unknown. Using transgenic mice that specifically expressed chemogenetic activators in their BF cholinergic neurons, we systematically investigated the role of BF cholinergic neurons in hypothyroidism-induced cognitive dysfunction by the combined approaches of patch clamp electrophysiology, behavioral testing, and immunohistochemistry. The results showed that LT4 treatment in the adult-onset hypothyroid mice reversed only 78 % of the BF cholinergic neurons to their normal values of electrophysiological properties. LT4 monotherapy did not rehabilitate cognitive function in the hypothyroid mice. Chemogenetic selective activation of the BF cholinergic neurons combined with LT4 treatment significantly improved learning and memory functions in the hypothyroid mice. In addition, chemogenetic activation of the cholinergic neurons induced the robust expression of c-Fos protein in the BF, prefrontal cortex (PFC), and hippocampus. This indicated that the BF cholinergic neurons improved learning and memory functions in the hypothyroid mice via the BF-PFC and BF-hippocampus pathways. In the hypothyroid C57BL/6 J mice, combined treatment via LT4 and donepezil, a cholinesterase inhibitor, significantly increased cognitive functions. The results suggested that the BF cholinergic neurons are critical for regulating learning and memory and reveal a novel pathophysiological mechanism for hypothyroidism-induced cognitive impairments.

[Analysis of Rapid Start-up and Mixed Nutritional Nitrogen Removal Performance of Complete Autotrophic Granular Sludge].

A rapid start of the completely autotrophic nitrogen removal over nitrite (CANON) process based on granular sludge and efficient nitrogen removal under mixotrophic conditions are important steps in a continuous flow reactor for CANON engineering applications. In this study, an aged CANON granular sludge was mechanically crushed to 0.3 mm as inoculum in an airlift internal-loop reactor (AIR) to achieve simultaneous COD removal and mixotrophic denitrification of the single-stage granular sludge. The system achieved stable partial nitrification by controlling DO after 26 days of startup. Granulation and anaerobic ammonia oxidation were then promoted by shortening the HRT to increase the ammonia nitrogen load to 5.65 kg ·(m3 ·d)-1. The total nitrogen removal rate reached 58% on the 68th day. Subsequently, the C/N ratio of influent was increased from 0 to 0.25 and 0.5, which promoted the synergistic growth of AOB, AMX, and heterotrophic microorganisms. The removal rates of ammonia and total nitrogen were 95% and 85% respectively, and the removal of COD reached approximately 80%. The activity of NOB such as Nitrospira was effectively inhibited as the COD concentration was increased. q(NH4+-N) and q(TN) were stable at 0.4 g ·(g ·h)-1 and 0.34 g ·(g ·h)-1, respectively, while q(NO3--N) was approximately 0.02 g ·(g ·h)-1. Microbial diversity was observed using MiSeq high-throughput sequencing. It showed that organic carbon had no significant effect on the abundance of Nitrosomomas and Candidutus_Kuenenia, while increasing the abundance of Candidutus_Brocadia and Denitratisoma in the sludge. This study provides ideas for the rapid start of continuous flow CANON granular sludge process to treat wastewater with low C/N ratio.

Combined application of extended depth of field imaging, image stitching and polarized microscopy techniques in identification of Spatholobus suberectus.

Objective: As traditional techniques for microscopic identification of Chinese medicines currently lack objective and high-quality reference images, here we developed a systemic procedure to be used in microscopic identification of Chinese medicines, which would lead to more objective, effective and accurate identification process. Methods: Spatholobi Caulis (Jixueteng in Chinese) was used as the specimen in the development of such procedure. Jixueteng samples were microscopically examined in bright- and dark-field microscopy. Microscopic images were obtained by regular, EDF, and image stitching techniques. Results: The microscopic images of the characteristics in pulverized Jixueteng were captured, thanks to EDF imaging and image stitching techniques which allowed the detailed and full sighting of each characteristic to be obtained simultaneously. Different layers in anatomical transverse section, including cork, phelloderm, cortex, phloem, cambium, xylem and pith, were distinctively observed. Moreover, by comparing images of bright- and dark-field microscopy, birefringent and non- birefringent components could readily be distinguished. Conclusion: With application of the developed procedure, high-definition, panoramic and microscopic images were acquired, which could be used as the reference images for microscopic identification of Chinese medicines.

A Fluorescent Coumarin-Based Probe for the Fast Detection of Cysteine with Live Cell Application.

A new coumarin-based fluorescent probe, containing an allylic esters group, has been designed and synthesized for sensing cysteine in physiological pH. In this fluorescent probe, the coumarin was applied as the fluorophore and an allylic esters group was combined as both a fluorescence quencher and a recognition unit. The probe can selectively and sensitively detect cysteine (Cys) over homocysteine, glutathione, and other amino acids, and has a rapid response time of 30 min and a low detection limit of 47.7 nM. In addition, the probe could be applied for cell imaging with low cytotoxicity.