Thyrotropin-Releasing-Hormone-Synthesizing Neurons of the Hypothalamic Paraventricular Nucleus are Inhibited by Glycinergic Inputs

Glycine is a classical neurotransmitter that has role in both inhibitory and excitatory synapses. To understand whether glycinergic inputs are involved in the regulation of the hypophysiotropic thyrotropin-releasing hormone (TRH) neurons, the central controllers of the hypothalamic-pituitary-thyroid axis, the glycinergic innervation of the TRH neurons was studied in the hypothalamic paraventricular nucleus (PVN). Methods: Double-labeling immunocytochemistry and patch-clamp electrophysiology were used to determine the role of glycinergic neurons in the regulation of TRH neurons in the PVN. Anterograde and retrograde tracing methods were used to determine the sources of the glycinergic input of TRH neurons. Results: Glycine transporter-2 (GLYT2), a marker of glycinergic neurons, containing axons were found to establish symmetric type of synapses on TRH neurons in the PVN. Furthermore, glycine receptor immunoreactivity was observed in these TRH neurons. The raphe magnus (RMg) and the ventrolateral periaqueductal gray (VLPAG) were found to be the exclusive sources of the glycinergic innervation of the TRH neurons within the PVN. Patch-clamp electrophysiology using sections of TRH-IRES-tdTomato mice showed that glycine hyperpolarized the TRH neurons and completely blocked the firing of these neurons. Glycine also markedly hyperpolarized the TRH neurons in the presence of tetrodotoxin demonstrating the direct effect of glycine. In more than 60% of the TRH neurons, spontaneous inhibitory postsynaptic currents (sIPSCs) were observed, even after the pharmacological inhibition of glutamatergic and GABAergic neuronal transmission. The glycine antagonist, strychnine, almost completely abolished these sIPSCs, demonstrating the inhibitory nature of the glycinergic input of TRH neurons. Conclusions: These data demonstrate that TRH neurons in the PVN receive glycinergic inputs from the RMg and the VLPAG. The symmetric type of synaptic connection and the results of the electrophysiological experiments demonstrate the inhibitory nature of these inputs

A Novel Pathway Regulates Thyroid Hormone Availability in Rat and Human Hypothalamic Neurosecretory Neurons

Hypothalamic neurosecretory systems are fundamental regulatory circuits influenced by thyroid hormone. Monocarboxylate-transporter-8 (MCT8)-mediated uptake of thyroid hormone followed by type 3 deiodinase (D3)-catalyzed inactivation represent limiting regulatory factors of neuronal T3 availability. In the present study we addressed the localization and subcellular distribution of D3 and MCT8 in neurosecretory neurons and addressed D3 function in their axons. Intense D3-immunoreactivity was observed in axon varicosities in the external zone of the rat median eminence and the neurohaemal zone of the human infundibulum containing axon terminals of hypophysiotropic parvocellular neurons. Immuno-electronmicroscopy localized D3 to dense-core vesicles in hypophysiotropic axon varicosities. N-STORM-superresolution-microscopy detected the active center containing C-terminus of D3 at the outer surface of these organelles. Double-labeling immunofluorescent confocal microscopy revealed that D3 is present in the majority of GnRH, CRH and GHRH axons but only in a minority of TRH axons, while absent from somatostatin-containing neurons. Bimolecular-Fluorescence-Complementation identified D3 homodimers, a prerequisite for D3 activity, in processes of GT1-7 cells. Furthermore, T3-inducible D3 catalytic activity was detected in the rat median eminence. Triple-labeling immunofluorescence and immuno-electronmicroscopy revealed the presence of MCT8 on the surface of the vast majority of all types of hypophysiotropic terminals. The presence of MCT8 was also demonstrated on the axon terminals in the neurohaemal zone of the human infundibulum. The unexpected role of hypophysiotropic axons in fine-tuned regulation of T3 availability in these cells via MCT8-mediated transport and D3-catalyzed inactivation may represent a novel regulatory core mechanism for metabolism, growth, stress and reproduction in rodents and humans

Dynamics of Bioactive Compounds during Spontaneous Fermentation of Paste Obtained from <i>Capsicum</i> ssp.—Stage towards a Product with Technological Application

Six cultivars of chili (Cherry, Bulgarian Chilli, Cayenne, Fatalii, Habanero, and Carolina Reaper) from two species (Capsicum annuum and Capsicum chinense) have been studied. Anaerobic, spontaneous fermentation of pure chili paste was conducted for 21 days at 20 °C. The unfermented (UCP) and fermented chili pastes (FCP) were both subjected to physicochemical and microbiological characterization consisting of capsaicinoid, ascorbic acid, short-chain organic acids, phenolic compounds, and simple sugars analysis. Cell viability for Lactic Acid Bacteria (LAB) and Leuconostoc was determined before and after fermentation. Results indicate that capsaicinoids are very stable compounds, as notable differences between unfermented and fermented samples could not be seen. Carolina Reaper and Fatalii cultivars were amongst the most pungent, whereas Cherry, Cayenne, and Bulgarian types were low to moderate in pungency. Average loss of total ascorbic acid was 19.01%. Total phenolic compounds ranged between 36.89–195.43 mg/100 g for the fresh fruits and 35.60–180.40 mg/100 g for the fermented product. Losses through fermentation were not significant (p 6–6.2 × 108 CFU/g) was high in all samples. Fermented chilies paste with enhanced biochemical and bacterial properties might further be used in the technology of vegetable (brining) or meat (curing) products, processes that generally involve the fermenting activity of different microorganisms, especially (LAB). Thus, the purpose of this research was the investigation of biochemical and microbial transformations that naturally occur in fermented chilies with a future perspective towards technological applications in cured meat products

Dynamics of Bioactive Compounds during Spontaneous Fermentation of Paste Obtained from Capsicum ssp.&mdash;Stage towards a Product with Technological Application

Six cultivars of chili (Cherry, Bulgarian Chilli, Cayenne, Fatalii, Habanero, and Carolina Reaper) from two species (Capsicum annuum and Capsicum chinense) have been studied. Anaerobic, spontaneous fermentation of pure chili paste was conducted for 21 days at 20 &deg;C. The unfermented (UCP) and fermented chili pastes (FCP) were both subjected to physicochemical and microbiological characterization consisting of capsaicinoid, ascorbic acid, short-chain organic acids, phenolic compounds, and simple sugars analysis. Cell viability for Lactic Acid Bacteria (LAB) and Leuconostoc was determined before and after fermentation. Results indicate that capsaicinoids are very stable compounds, as notable differences between unfermented and fermented samples could not be seen. Carolina Reaper and Fatalii cultivars were amongst the most pungent, whereas Cherry, Cayenne, and Bulgarian types were low to moderate in pungency. Average loss of total ascorbic acid was 19.01%. Total phenolic compounds ranged between 36.89&ndash;195.43 mg/100 g for the fresh fruits and 35.60&ndash;180.40 mg/100 g for the fermented product. Losses through fermentation were not significant (p &lt; 0.05). Plate counts indicated low initial numbers for LAB in the fresh samples, values ranging between 50&ndash;3700 CFU/g (colony-forming units). After fermentation, day 21, concentration of LAB (3.8 &times; 106&ndash;6.2 &times; 108 CFU/g) was high in all samples. Fermented chilies paste with enhanced biochemical and bacterial properties might further be used in the technology of vegetable (brining) or meat (curing) products, processes that generally involve the fermenting activity of different microorganisms, especially (LAB). Thus, the purpose of this research was the investigation of biochemical and microbial transformations that naturally occur in fermented chilies with a future perspective towards technological applications in cured meat products

Advanced Characterization of Hemp Flour (Cannabis sativa L.) from Dacia Secuieni and Zenit Varieties, Compared to Wheat Flour

The advanced characterization of flour from hemp seeds (edible fruits of Cannabis sativa L.) from the Dacia Secuieni and Zenit varieties, compared to wheat flour, was studied in this research. The aim was to present the characterization of 2 varieties, out of the 70 accepted in Europe, for human consumption. The varieties selected from hemp meet the THC level requirement (0.3 or 0.2% of the dry weight of the reproductive part of the female flowering plant) in seeds. Hemp flour was obtained by grinding. The flour samples were evaluated for physicochemical parameters (moisture, crude protein, lipids, ash, crude fiber), the content of micro and macro elements, fatty acids, amino acids, and carbohydrates. The total proteins in hemp flour are found in larger quantities by over 35% compared to wheat flour, and the lipids reach the threshold of 28%. The amount of mineral substances exceeds 3% in hemp flour, and the fibers exceed 26%, compared to 0.61% for wheat flour. The predominant mineral substances were K, Ca, Mg, p, Fe, and Mn. The predominant fatty acids were the unsaturated ones, predominantly being linoleic acid, followed by oleic and gamma-linoleic acid. In the case of amino acids, the highest amount is found in glutamic acid for hemp flours. As for carbohydrates, sucrose is found in the largest amount, followed by glucose and fructose. In conclusion, hemp flours have superior non-traditional characteristics to wheat flour, being a potential raw material for fortifying food or using them as such, having beneficial effects of consumption on the proper functioning of the human body

Freeze-Dried Powder of Fermented Chili Paste—New Approach to Cured Salami Production

Fermented chili powders were obtained through the freeze-drying of fermented chili pastes and used as a condiment, acidifier, antioxidant, colorant, and microbial starter carrier in fermented salami production. Fermented chili powders were examined regarding carbohydrates, organic acids, vitamin C, phenolic compounds, carotenoids, and aroma profile. High concentrations of lactic (10.57–12.20%) and acetic acids (3.39–4.10%) were recorded. Vitamin C content was identified in the range of 398–1107 mg/100 g, with maximum values for C. annuum cv. Cayenne chili powder. Phenolic compounds showed values between 302–771 mg/100 g. Total carotenoid content was identified between 544–2462 µg/g, with high concentrations of capsanthin esters. Aroma profile analysis evidenced specific compounds (1-hexanol, 2-hexanol, hexenal, E-2-hexenal) with sensory importance and a more complex spectrum for Capsicum chinense cultivar. Plant-specific lactic acid bacteria showed dominance both in fermented chili paste, chili powder, and salami. Lactic and acetic acids from the fermented chili powder reduced the pH of the filling immediately, having a stabilizing effect on the meat. Nor molds or pathogens were identified in outer limits. Based on these results, fermented chili powders could be used as starter carriers in the production of fermented meat products for exceptional sensory properties and food safety management