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Hypoglycemia deprives the brain of its primary energy source glucose. Reductions in whole-brain amino acid energy substrate levels suggest that these non-glucose fuels may be metabolized during glucose shortage. Recurring hypoglycemia can cause mal-adaptive impairment of glucose counter-regulation; yet, it is unclear if amplified reliance upon alternative metabolic substrates impedes detection of continuing neuro-glucopenia. This research aimed to develop high-sensitivity UHPLC-electrospray ionization mass spectrometric (LC-ESI-MS) methodology, for complementary use with high-neuroanatomical resolution microdissection tools, for measurement of glucogenic amino acid, e.g. glutamine (Gln), glutamate (Glu), and aspartate (Asp) content in the characterized glucose-sensing ventromedial hypothalamic nucleus (VMN) during acute versus chronic hypoglycemia. Results show that VMN tissue Gln, Glu, and Asp levels were significantly decreased during a single hypoglycemic episode, and that Gln and Asp measures were correspondingly normalized or further diminished during renewed hypoglycemia. Results provide proof-of-principle that LC-ESI-MS has requisite sensitivity for amino acid energy substrate quantification in distinctive brain gluco-regulatory structures under conditions of eu- versus hypoglycemia. This novel combinatory methodology will support ongoing efforts to determine how amino acid energy yield may impact VMN metabolic sensory function during persistent hypoglycemia. Untargeted mass spectrometry analysis is one of the most challenging and meaningful steps in the rapid structural elucidation of the highly complex and diverse constituents of traditional Chinese medicine. Specifically, it is a laborious and time-consuming way to identify unknown compounds. Herein, a workflow was proposed to expedite the annotations of the chemical structures in Pheretima aspergillum (E. Perrier) (Di-Long, DL). First, ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOFMS) was performed to obtain the untargeted mass spectral data. Then, the spectral data were uploaded to the Global Natural Products Social Molecular Networking (GNPS) platform to create a network and extract the Mass2Motifs (co-occurring fragments and neutral losses) using unsupervised substructure annotation topic modeling (MS2LDA). Finally, a structural analysis was performed using the proposed workflow of MS2LDA in combination with mass spectral molecular networking and in silico fragmentation prediction. As a result, a total of 124 compounds from DL were effectively characterized, of which 89 (7 furan sulfonic acids, 57 phospholipids and 25 carboxamides) were identified as potentially new compounds from DL. The results presented in this article significantly improve the understanding of the chemical composition of DL and provide a solid scientific basis for the future study of the quality control, underlying pharmacology and mechanism of DL. Moreover, the proposed workflow was used for the first time to accelerate the annotations of unknown molecules from TCM. Furthermore, this workflow will increase the efficiency of characterizing the 'unknown knowns' and elucidation of the 'unknown unknowns' from TCM, which are crucial steps of discovering the natural product drugs in TCM. V.The present study investigated whether or not passive immunization against inhibin modulates testicular blood flow in goats. Male Shiba goats were injected with either 10 ml of inhibin antiserum (INH group; n = 5) or 10 ml of normal castrated goat serum (NGS group; n = 4). Concentrations of FSH, LH, testosterone (T), and estradiol (E2) in the plasma were measured by radioimmunoassay. Blood flow within the supratesticular (STA) and marginal testicular arteries (MTA) were measured by color pulsed-Doppler ultrasonography, and Doppler indices (resistive index; RI and pulsatility index; PI) were recorded. Results revealed significant increases in concentrations of FSH and E2 in the INH group compared to those in the NGS group (P  less then  0.05). Animals in the INH group had greater (P  less then  0.05) FSH concentrations than those in the NGS group in the period between 60 h and 144 h after treatment than at any other time. Estradiol concentrations were greater (P  less then  0.05) in the INH group than in the NGS group at 6 h (12.15 ± 2.09 pg/ml vs 5.49 ± 1.17 pg/mL), 12 h (8.27 ± 1.29 pg/mL vs 3.05 ± 0.38 pg/mL), and 36 h (9.35 ± 1.31 pg/mL vs 5.09 ± 0.46 pg/mL) after treatment than at any other time. Concentrations of LH and T did not significantly change between the two groups. Goats in the INH group had lesser (P  less then  0.05) RI of the STA than those in the NGS group and RI values were lesser at 24 h (0.37 ± 0.031 vs 0.49 ± 0.004) and 120 h (0.38 ± 0.028 vs 0.55 ± 0.048) after treatment than at any other time. Furthermore, values of RI and PI of the MTA were significantly lesser (P  less then  0.05) in the INH group compared to those in the control group at 48 h (RI of MTA 0.21 ± 0.014 vs 0.37 ± 0.039; PI of MTA 0.24 ± 0.016 vs 0.46 ± 0.058) after treatment than at any other time. Q-VD-Oph chemical structure In conclusion, passive immunization against inhibin has a stimulatory effect on testicular blood flow in goats by inducing decreases in the RI values of the STA and MTA. The effect of body condition and environmental contaminants on reproductive processes is known; however, it is not known whether basic ovarian cell functions and their response to these contaminants depend on body condition. This study aimed to understand the interrelationships between body conditions and environmental contaminants on ovarian cells. For this purpose, we compared ovarian granulosa cells isolated from cows with an emaciation tendency (body condition score, BCS2 on a scale from 1 to 5) and cows with average body condition (BCS3); proliferation, apoptosis, secretory activity and the response to environmental contaminants were all assessed in the cells. In the 1st series of experiments, ovarian granulosa cells isolated from BCS2 and BCS3 cows were cultured with and without benzene, xylene, and toluene (0.1%). The accumulation of nuclear and cytoplasmic markers of apoptosis (p53 and bax, respectively), a proliferation marker (PCNA), progesterone (P4), and insulin-like growth factor I (IGF-I) was evaluated by Western blot and radioimmunoassay (RIA) experiments.