Mycobacterial Populations Partly Change the Proportions of the Cells Undergoing Asymmetric/Symmetric Divisions in Response to Glycerol Levels in Growth Medium

Twenty to thirty percent of the septating mycobacterial cells of the mid-log phase population showed highly deviated asymmetric constriction during division (ACD), while the remaining underwent symmetric constriction during division (SCD). The ACD produced short-sized cells (SCs) and normal/long-sized cells (NCs) as the sister–daughter cells, but with significant differential susceptibility to antibiotic/oxidative/nitrite stress. Here we report that, at 0.2% glycerol, formulated in the Middlebrook 7H9 medium, a significantly high proportion of the cells were divided by SCD. When the glycerol concentration decreased to 0.1% due to cell-growth/division, the ACD proportion gradually increased until the ACD:SCD ratio reached ~50:50. With further decrease in the glycerol levels, the SCD proportion increased with concomitant decrease in the ACD proportion. Maintenance of glycerol at 0.1%, through replenishment, held the ACD:SCD proportion at ~50:50. Transfer of the cells from one culture with a specific glycerol level to the supernatant from another culture, with a different glycerol level, made the cells change the ACD:SCD proportion to that of the culture from which the supernatant was taken. RT-qPCR data showed the possibility of diadenosine tetraphosphate phosphorylase (MSMEG_2932), phosphatidylinositol synthase (MSMEG_2933), and a Nudix family hydrolase (MSMEG_2936) involved in the ACD:SCD proportion-change in response to glycerol levels. We also discussed its physiological significance.

Piceatannol, a Natural Stilbene, Extends the Lifespan of Caenorhabditis elegans Under Fasting Through Inhibition of Lipolysis

Objectives Lipolysis is the catabolic process that hydrolyzes triglyceride (TG) to free fatty acids (FFAs) and glycerol under negative energy balance such as fasting. In adipocytes, adipose TG lipase (ATGL), hormone-sensitive lipase (HSL), and monoglyceride lipase play key roles in a series of TG hydrolysis reactions in mammals. However, overly activated adipose lipolysis is believed to contribute to link between obesity and systemic inflammation and oxidative stress. We previously demonstrated that piceatannol (PIC), a natural resveratrol analogue, inhibits adipogenesis in cultured adipocytes and lipogenesis in Caenorhabditis elegans. Furthermore, we showed that PIC extends the lifespan of C. elegans via the insulin/IGF-1 signaling. However, the effects of PIC on lipid metabolism during fasting state is unknown. Methods We conducted Oil-Red-O assay, Enzyme assay (TG and Free glycerol contents), PCR analysis and lifespan assay. Results In this study, we demonstrated that PIC-treated C. elegans exhibited suppressed lipolysis under fasting as judged by increased lipid accumulation and TG levels with decreased free glycerol level. Consistent with these findings, PIC treatment resulted in decreased mRNA levels of genes involved lipolysis such as atgl-1, hosl-1 and aak-2 in fasted C. elegans. Also, PIC treatment augmented fasting-induced lifespan of C. elegans by an increased daf-16 gene expression. However, such effect was abolished when atgl-1, aak-2, and daf-16 mutants were treated with PIC. In addition, we also found that autophagy is required for PIC-induced lifespan in C. elegans during fasting since autophagy inhibitor treatments and autophagy gene deficient worms resulted in blunting the lifespan extension effect of PIC. Conclusions Collectively, our results indicate that PIC contributes to lifespan extension in C. elegans during fasting possibly through regulating lipolysis- and/or autophagy-dependent lipid metabolism. Funding Sources 1. The National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (2019R1A2C1086146) and (2019R1A6A3A03033878) 2. The Rural Development Administration of the Republic of Korea.

Spurious hypertriglyceridaemia in unconscious patient

Background: Routine biochemical investigation of one patient admitted with a diagnosis of CVA as mentioned in the requisition revealed high hypertriglyceridaemia in a non-lipemic serum for the first time. Later, after scrutiny of the case sheet in the ward, it was found that apart from other management, blood was drawn about 90 minutes after administration of 4th dose of oral glycerol through nasogastric tube and sent to the laboratory. This was suspected to be the probable cause. In order to find out the degree of interference in blood sample, a small amount of glycerol was brought from the ward to experiment.Methods: Glycerol was initially diluted to 1 in 100 in distilled water and then artificially mixed in various dilutions with pooled serum from indoor patients and pooled heparinized plasma from outdoor patients. These samples were subjected to triglycerides estimation while routine analyses were going on.Results: A concentration of about 13,50,500 mg/dl falsely measured triglycerides was found in glycerol solution. This needed a dilution of 1 in 1500 for distilled water, serum and plasma which had 0, 190 and 113 mg/dl triglycerides respectively to bring the level to near the highest range of linearity. The recovery of added glycerol in distilled water was almost 100% but there was some positive bias more with plasma than serum.Conclusions: The spurious hypertriglyceridaemia in serum resulted from positive interference due to temporary high glycerolaemia by the commonly employed GPO-PAP method for its measurement and would cause the same depending on glycerol level.

Functional and Film-forming Properties of Mechanically Deboned Chicken Meat Proteins

In the present study, the functional properties of mechanically deboned chicken meat proteins (MDCM-Ps) were determined at different pH (2, 4, 6, 8 and 12), and film-forming properties were evaluated depending on MDCM-P (2, 3 and 4 %) and glycerol (30, 40 and 50 %) concentrations. The highest solubility, emulsifying and foaming properties were determined at pH 12. MDCM-P solutions showed non-polyelectrolyte behavior and gelation onset temperature was determined at 36 °C. Film-forming properties of MDCM-P showed that the tensile strength decreased, and elongation at break increased as glycerol concentration increased. Films became more transparent with increasing glycerol and decreasing protein concentration, while water vapor permeability increased with increasing glycerol and protein concentration. Water sorption data of films were fitted to the Guggenheim, Anderson, and De Boer model. In general, equilibrium moisture content of films increased as glycerol level increased. Overall, the results showed that MDCM-P could be useful as a new protein source for both food and packaging industries.

THE PROPERTIES OF EDIBLE FILM DERIVED FROM BOVINE SPLIT HIDE GELATIN WITH ISOLATED SOY PROTEIN USING VARIOUS LEVELS OF GLYCEROL IN THE PRESENCE OF TRANSGLUTAMINASE

This study examined the characteristics of the edible film mixture gelatin bovine split hide and soy protein isolate, cross-linked using transglutaminase enzyme with addition level of glycerol as a plasticizer.The material used gelatin combination (Gelatin bovine split hide: soy proteinisolate = 90: 10 with 30 Utransglutaminase enzyme levels ), and glycerol. Data were analyzed using completely randomized design each treatment was replicated thrice of glycerol as a treatment level ( 10%, 20% dan 30%). The results showed that the glycerol level significantly affected the thickness, elongation, solubility, degree of cross-linking and WVTR, but not the tensile strength.The addition of glycerol causing the film becomes more elastic and the surface is more homogeneous and compact. Increasing the level to 30% glycerol affects the mechanical properties of an edible film.

Are Primary Supratentorial Intracerebral Hemorrhages Surrounded by a Biochemical Penumbra? A Microdialysis Study

OBJECTIVE: Opinions vary regarding the indications for surgical evacuation of spontaneous intracerebral hemorrhages (ICH) and whether or not penumbra zones surround them. METHODS: We performed intracerebral microdialysis (mean duration, 3.5 d) after surgical evacuation of ICH in 22 patients. Probes were placed in the parenchyma within 1 to 2 cm of the evacuated hematoma; a postoperative computed tomographic scanning verified their positions. The catheters were perfused with an artificial cerebrospinal fluid solution at 0.3 μl/min. Biochemical variables (glucose, pyruvate, lactate, glutamate, and glycerol) were analyzed and displayed at the bedside. The levels obtained were compared with previous data from normal human brains and the pericontusional penumbra zones of patients with severe traumatic brain lesions. RESULTS: During 1 to 12 hours after surgery, interstitial levels of glucose (median level, 1.3 mmol/L; interquartile range, 0.6–2.2 mmol/L) were within normal variations, whereas the levels of lactate (median level, 6.4 mmol/L; interquartile range, 3.9–9.0 mmol/L), glutamate (median level, 14 μmol/L; interquartile range, 5–370 μmol/L), and glycerol (median level, 190 μmol/L; interquartile range, 74–380 μmol/L), as well as the lactate/pyruvate ratio (median ratio, 35; interquartile range, 23–50) were increased. A gradual normalization of the lactate/pyruvate ratio and glycerol level was observed within 48 hours. CONCLUSION: The area close to an evacuated ICH exhibits a biochemical pattern similar to that of the biochemical penumbra zone surrounding focal traumatic brain contusions. The presence of a penumbra zone around large ICH may be of importance for making surgical decisions.