Plasma lipoprotein profiles and arylesterase activities in two inbred strains of rabbits with high or low response of plasma cholesterol to dietary cholesterol

• 1.1. Cholesterol feeding for 4 weeks of female and male rabbits of two inbred strains increased plasma cholesterol concentrations by about 11 and 48 mmole/I in the hypo- and hyperresponsive strain, respectively.
• 2.2. On the low-cholesterol pre-experimental diet, the hyporesponsive animals had significantly higher plasma HDL (high density protein) cholesterol levels than hyperresponders.
• 3.3. In both strains, cholesterol feeding caused elevations of cholesterol in all lipoprotein classes, the difference between the hypo- and hyperresponsive strains in essence only being observed in the VLDL (very low density lipoprotein) fraction.
• 4.4. Basal plasma total arylesterase activity was significantly higher in the hypo- than in the hyperresponsive rabbits.
• 5.5. Dietary cholesterol caused an increase in plasma esterase activity in both strains.
• 6.6. We suggest that in rabbits a low plasma arylesterase activity and a low concentration of HDL cholesterol are associated with an increased sensitivity to dietary cholesterol.

     

Glucosyl and galactosyl transferase activities of diabetic (db/db) and obese (ob/ob) mice kidneys

• 1.1. Glucosyl and galactosyl activities were determined in kidney cortex tissue prepared from two strains of mice, genetically diabetic and obese mice.
• 2.2. These activities were measured as a function of ageing between 6 weeks and 13 months.
• 3.3. For both strains glucosyl transferase activity was shown to increase with respect to ageing whereas galactosyl transferase activity decreased at the same time.
• 4.4. These changes of enzymatic activities would suggest that a smaller increase of hydroxylysine-linked glycans than expected was observed under these pathological conditions.

     

Differential vitamin and choline requirements of symbiotic and aposymbiotic s. oryzae (coleoptera: curculionidae)

• 1.1. Development times, fertilities and weights of symbiotic and aposymbiotic strains of the rice weevil Sitophilus oryzae in response to single omissions of seven vitamins and choline were studied.
• 2.2. No strict requirement for choline could be demonstrated: development times and weights were comparable to diets with or without choline, but the presence of choline increased fertility of the symbiotic strain on vitamin-deficient diets.
• 3.3. The 0.02% concentration used in these experiments may be too high since it resulted in a lower fertility of the aposymbiotic strain.
• 4.4. The results indicated that niacin could be synthesized by the weevils.
• 5.5. Thiamine, folic acid and pyridoxine were found to be required in the diet of both strains but the latter two may be supplied in part by symbiotes since a first symbiotic generation was obtained.
• 6.6. Pantothenic acid, biotin and riboflavin were only required by aposymbiotic weevils: their omission from the diet did not modify the development of symbiotic insects.
• 7.7. It is therefore suggested that these three vitamins are supplied by symbiotes.

     

Distribution of serine esterase activity in the lymphoid system of C57bl/6 mice,effect of aging on the enzymatic activity

• 1.1. The activity of serine esterase (SE) was investigated in the lymphoid system of C57BL/6 mice. SE activity increased in the lymphoid tissues with their content of mature T-lymphocytes, except that high levels were also observed in various populations of bone marrow cells.
• 2.2. The maturation of T-lymphocytes in the thymus was accompanied by an increase in their SE activity.
• 3.3. Experiments on the influence of age on SE activity showed that while thymocytes were not affected, a three-fold increase in activity occurs in spleen lymphocytes between the ages of 26 and 78 wk.

     

Proteomic Analysis of Baboon Cerebral Artery Reveals Potential Pathways of Damage by Prenatal Alcohol Exposure,

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Highlights

• •We studied mid-pregnancy alcohol exposure and baboon fetal cerebral artery.
• •238 proteins differed between control and alcohol-exposed fetuses near-term.
• •Proteins of metabolic pathways represented one of the major targets of alcohol.
• •Alcohol effect on the development of fetal brain vessels is persistent.

     

Isohematoporphyrin-diesters and diethers

• 1.1. A series of diesters of isohematoporphyrin (isoHp), from dimethyl to dioctyl were prepared according to Rimington et al. (1989b). Their optical absorption, fluorescence spectra and high performance liquid chromatography (HPLC) retention times were recorded.
• 2.2. A plot of HPLC retention time against number of C atoms in the alcohol used for esterification was approximately linear at first then rising steeply from diamyl to diocyi ester, whether a gradient elution was used or only methanol: water, 95/5, at pH 7.5.
• 3.3. Preparation of the diethers of isoHp was much more difficult than that of the corresponding derivatives of hematoporphyrin (Hp). Several different methods were investigated, varying both times and temperatures.
• 4.4. These methods included reaction of isoHp or its demethyl ester with
  • 4.1.(i) a bromoalkane in presence of anhydrous K₂CO₃;
  • 4.2.(ii) reaction with bromoalkane and Ag₂O;
  • 4.3.(iii) reaction of brominated-isoHp, prepared by using thionylbromide, with the selected alcohol, or corresponding sodium alcoholate;
  • 4.4.(iv) heating of isoHp alone with an alcohol containing 20% (w/v) H₂SCO₄ (temp. range from 45° to 118°C),
  • 4.5.(v) refluxing as in (iv) at the b.p. of the alcohol; and
  • 4.6.(vi) carrying out this reaction in refluxing ethyleneglycoldimethyl ether (b.p. 85°C) or diethyleneglycoldimethyl ether (b.p. 155°C).
• 5.5. Some diether formation was observable by all these methods but yields were small, a considerable quantity of unreacted isoHp and other products remaining.
• 6.6. Examined by HPLC, the diethers consistently afforded a forked peak which on thin layer chromatography was only resolved into two very closely associated bands by a solvent mixture carefully selected for development.
• 7.7. On elution these materials had virtually identical optical absorption and fluoresence spectra.
• 8.8. The nature of the association is discussed, atropisomers (Gottwald and Ullman, 1969) and possible stacked monomer: dimers (Abraham et al., 1963) being considered as possibilities.

     

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The effects of chronic metabolic acidosis on patterns of protein synthesis in rat renal cortex

• 1.1. In the rat chronic metabolic acidosis increases the net synthesis of 17 renal cortex proteins by amounts ranging from 1.5 to 4.5-fold.
• 2.2. These proteins have molecular weights between 13,000 and 42,000 and isoelectric points between approximately 5.5 and 7.0.
• 3.3. No new proteins not also present in normal animals are detected in renal cortex samples from acidotic animals.
• 4.4. Three proteins undergo substantial reductions in their net synthetic rates in chronic metabolic acidosis.
• 5.5. On the basis of their physical properties and similar alterations in net synthetic rate in acidosis some of these proteins appear to be closely related and may be coordinately expressed in the rat kidney.

     

Purification and partial characterization of 200 kDa oncofetal antigen from radiation induced murine lymphocytic lymphoma

• 1.1. A 200 kDa glycoprotein (gp200) oncofetal antigen was purified from solubilized membranes of a radiation-induced murine lymphomcytic lymphoma cell line (XR11-5T), grown in syngeneic RFM mice, by successive gel chromatography of the active fraction on lentil lectin agarose, Q- and S-Sepharose and Superose-12 using an FPLC system.
• 2.2. A murine monoclonal antibody 115, produced by the syngeneic immunization of adult male C57BL/6N mice with 12-day mouse fetal cells, was used in a slot blot antibody assay to follow up the active fractions.
• 3.3. The purified glycoprotein has a pi of 5.4.
• 4.4. Treatment of radiolabeled gp200 with neuraminidase caused a slight reduction in size due to the removal of sialic acid groups and a shift in pI to 6.3.
• 5.5. Treatment of gp200 with different glycosidases shows that gp200 is susceptible to N- and O-glycanase but not to endoglycosaminidase H.
• 6.6. On extraction of gp200 with Triton X-114 it partitions exclusively into the detergent-rich fraction consistent with being an integral membrane protein.

     

Quantitative Microproteomics Based Characterization of the Central and Peripheral Nervous System of a Mouse Model of Krabbe Disease

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Highlights

• •Quantitative microproteomics to study the CNS and PNS of the Twitcher mouse.
• •10plex TMT experiments on corpus callosum, motor cortex and sciatic nerves extracts.
• •More than 400 proteins groups deregulated between Twitcher and wildtype mice.
• •New insights into the molecular mechanisms of Krabbe disease.

     

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Disruption of endothelial barrier function: relationship to fluidity of membrane extracellular lamella

• 1.1. Endothelial cells were cultured in tissue culture flasks or on microcarrier beads and labeled with a lipid specific spin-label.
• 2.2. Exposure of endothelial cells to benzyl alcohol caused a dose- and time-dependent increase in membrane fluidity using electron spin resonance (ESR). Maximum fluidity was reached after a 5-min exposure to 100 mM benzyl alcohol.
• 3.3. Albumin permeability across endothelial cells cultured on micropore filters was used as an indication of endothelial monolayer integrity.
• 4.4. A significant increase in permeability occurred with 50 mM benzyl alcohol. Maximal albumin permeability was reached after a 5-min exposure to 100 mM benzyl alcohol.

     

Exploring Regulation of Protein O-Glycosylation in Isogenic Human HEK293 Cells by Differential O-Glycoproteomics

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Highlights

• •A panel of HEK293 isogenic cell lines with knockout of GALNT genes.
• •Identification of nonredundant O-glycosylation sites regulated by specific GalNAc-T isoforms.
• •GalNAc-T7 and T10 contribute to follow-up activity in regions of high density O-glycosylation.
• •GalNAc-T11 specifically controls O-glycosylation of specific linker regions in the low-density lipoprotein receptor related proteins.

     

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Quantitative Proteomics of the 2016 WHO Neisseria gonorrhoeae Reference Strains Surveys Vaccine Candidates and Antimicrobial Resistance Determinants

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Highlights

• •Quantitative proteomes of the 2016 WHO Neisseria gonorrhoeae reference strains.
• •Novel gonorrhea vaccine candidates and potential global proteomic AMR markers.
• •First large-scale proteomic profiling of gonorrhea vaccine candidates and AMR.
• •A reference proteomics databank for gonococcal vaccine and AMR research endeavors.

     

Metabolism of secondary alcohols in Drosophila melanogaster: Effects on alcohol dehydrogenase

• 1.1. In vivo metabolism of a secondary alcohol in Drosophila melanogaster and its effects on alcohol dehydrogenase (ADH) have been studied.
• 2.2. ADH-mediated breakdown of the secondary alcohol, propan-2-ol, was the main source of the acetone produced.
• 3.3. Acetone formation declined and stopped ultimately, suggesting inhibition of ADH activity in vivo which has been confirmed in in vitro studies.
• 4.4. A powerful ketone-trapping agent, semicarbazide, did not restore the ADH activity in vitro, whereas aldehyde substrates of ADH did restore activity.
• 5.5. The final formation of a dead-end ADH:NAD-acetone ternary complex has been proposed and its consequences discussed.

     

1H NMR study of metabolic alterations in the small intestine of rats infected with Hymenolepis diminuta

• 1.1. 1H NMR spectra of the duodenum, jejunum and ileum tissues of the small intestine of a rat showed metabolic gradients.
• 2.2. The concentrations of metabolites in these gut regions were altered by the presence of the tapeworm Hymenolepis diminuta.
• 3.3. In the infected duodenum there was significantly less glycogen, glucose and phosphocreatine/creatine, but significantly more lactate than in the corresponding controls.
• 4.4. Infected jejunum contained significantly less betaine but significantly more succinate, alanine and lactate.
• 5.5. Infected ileum had significantly less glycogen and taurine but significantly more alanine and lactate.

     

Relationship between ecg,eeg and sps responses during arousal in the goldfish (Carassius auratus)

• 1.1. Goldfish (Carassius auratus) were fitted with ECG electrodes and intra-cranial stainless steel electrodes to monitor cardiac, EEG and SPS responses, during minimal restraint, to presentation of environmental stimuli (light-on, moving shadow, tap).
• 2.2. All three stimuli evoked a bradycardia and increases in the EEG frequency; correlates of arousal responses in fish.
• 3.3. EEG frequency changes were most evident in the fore- and midbrain regions; in the hindbrain smaller responses nevertheless showed discrimination between stimuli.
• 4.4. EEG amplitude changes were more site- and stimulus-specific than frequency changes.
• 5.5. SPSs occurred on stimulus presentations which were negative in polarity in the midbrain and positive in the forebrain and hindbrain, though the latter were smaller and less consistent.
• 6.6. Principal components analyses and regression analyses were used to examine detailed associations between peripheral and central physiological changes.
• 7.7. It was found that increases in the EEG frequency of fore- and midbrain regions were related to cardiac deceleration on early stimulus presentations.
• 8.8. This was also shown for the negative SPS of the midbrain to the presentation of the tap stimulus.
• 9.9. Positive SPSs of the forebrain were related to the bradycardia on later stimulus presentation i.e. during habituation of the arousal response.
• 10.10. The results indicate that in fish, as in other vertebrates, negative SPSs in the midbrain are associated with arousal and implicate the forebrain in the modulation of arousal by its habituation.