Purification of turkey pancreatic phospholipase A2

Turkey pancreatic phospholipase (TPP) has been purified from delipidated pancreases. The purification included ammonium sulfate fractionation, acidic (pH 5) treatment, followed by sequencial column chromatographies on DEAE-cellulose, Sephadex G-75, and reverse phase high pressure liquid chromatography. The purified enzyme was found to be a monomeric protein with molecular mass of 14 kDa. The optimal activity was measured at pH 8 and 37 degrees C using egg yolk emulsion as substrate. Our results show that the enzyme (TPP) was not stable for 1 h at 60 degrees C, and that bile salt and Ca2+ were required for the expression of the purified enzyme. The sequence of the N-terminal amino acids of the purified enzyme shows a very close similarity between TPP and all other known pancreatic phospholipases.     

Effect of time of day on line-bisecting performance in female soccer players

The purpose of this study was to examine the effect of time of day on visuospatial attention in trained subjects. We used the line-bisecting task. The protocol involved 30 healthy volunteer female soccer players (age: 21.1 ± 2.1 years, level of experience: 8.1 ± 3.2 years, height: 1.7 ± 0.03 cm, weight: 59.2 ± 3.49 kg). They performed two-test sessions at different times of day, 8:00 am and 8:00 pm. The results showed a significant time-of-day effect on line-bisecting performance, with a better precision in the evening than in the morning. The rightward error observed in the morning was reversed in the evening. An interaction between time of day x length was also observed, indicating that the difference in performance between the morning and the evening increased with the length of the line. We conclude that activities that require better visuospatial performance should be programed to take place in the evening.     

Molecular genetic analysis of the 17p11.2 region in patients with hereditary neuropathy with liability to pressure palsies (HNPP)

Hereditary neuropathy with liability to pressure palsies (HNPP) is in most cases associated with an interstitial deletion of the same 1.5-Mb region at 17p11.2 that is duplicated in Charcot-Marie-Tooth type 1A (CMT1A) patients. Unequal crossing-over following misalignment at flanking repeat sequences (CMT1A-REP), either leads to tandem duplication in CMT1A patients or deletion in HNPP patients. With the use of polymorphic DNA markers located within the CMT1A/HNPP duplication/deletion region we detected the HNPP deletion in 16 unrelated HNPP patients, 11 of Belgian and 5 of French origin. In all cases, the 1.5-Mb size of the HNPP deletion was confirmed by EcoRI dosage analysis using a CMT1A-REP probe. In the 16 HNPP patients, the same 370/320-kb EagI deletion-junction fragments were detected with pulsed field gel electrophoresis (PFGE), while in CMT1A patients, a 150-kb EagI duplication-junction fragment was seen. Thus, PFGE analysis of EagI-digested DNA with a CMT1A-REP probe allows direct detection of the HNPP deletion or the CMT1A duplication for DNA diagnostic purposes.     

Chemical Composition and Bioactivities of Quercus canariensis Flour Acorns Extracts Growing in Tunisia

The current study was conducted to investigate the chemical composition of Quercus canariensis flour acorns extracts as well as its biological activities in regards to the growing area using spectrophotometric and chromatographic techniques. The phenolic profile was composed of 19 compounds identified through HPLC-DAD analysis. Coumarin was the most abundant compound quantified in BniMtir, Nefza and ElGhorra and gallic (12.58–20.52 %), syringic (4.70–7.64 %) and trans-ferulic (2.28–2.94 %) acids were the abundant phenolic acids while kaempferol was the major flavonoid compounds quantified only in Quercus canariensis growing in BniMtir. On the other hand, Ain Snoussi acorn extract was characterized by its high content in luteolin-7-O-glucoside (58.46 %). The in-vitro antioxidant activities of the studied extracts were investigated and the results showed that Nefza ethanolic extract's has the highest activities. A bactericidal effect against Staphylococcus aureus was observed only by Elghorra population. On the other hand, Ain Snoussi acorn extract was efficient to inhibit growth of pathogenic bacteria, mentioned the highest activity against Escherichia coli. These results is the first study highlighted that zeen oak acorns are an excellent source of natural antioxidants and antibacterial compounds related to their lysozyme activity which could be exploited in the pharmaceutical and food sectors.     

Lipid peroxidation,antioxidant activities and stress protein (HSP72/73, GRP94) expression in kidney and liver of rats under lithium treatment

The present work was aimed at studying the effects of a subchronic lithium treatment on rat liver and kidneys, paying attention to the relationship between lithium toxicity, oxidative stress, and stress protein expression. Male rats were submitted to lithium treatment by adding 2 g of lithium carbonate/kg of food for different durations up to 1 month. This treatment led to serum concentrations ranging from 0.5 mM (day 7) to 1.34 mM (day 28) and renal insufficiency highlighted by an increase of blood creatinine and urea levels and a decrease of urea excretion. Lithium treatment was found to trigger an oxidative stress both in kidney and liver, leading to an increase of lipid peroxidation level (TBARS) and of superoxide dismutase and catalase activities. Conversely, glutathione peroxidase activity was reduced. Constitutive HSP73 (heat shock protein 73) expression was not modified by lithium treatment, whereas inducible HSP72 was down-regulated in kidney. GRP94 (glucose regulated protein 94) appeared as two isoforms of 92 and 98 kDa: the 98-kDa protein being overexpressed in kidney by lithium treatment whereas 92-kDa protein was underexpressed both in kidney and liver.     

Pharmacokinetics, immunogenicity, and efficacy of dimeric TNFR binding proteins in healthy and bacteremic baboon

Immunogenicity, pharmacokinetics, and therapeutic efficacy ofthree novel dimeric soluble tumor necrosis factor (TNF)-receptor Iconstructs [TNF-binding protein (bp)] were evaluated in 28 baboons, 12 of which were healthy and 16 were challenged with a lethal Escherichia coli bacteremia. The threeconstructs differed only in the number of extracellular domains of theTNF receptor I and were dimerized with polyethylene glycol. Althoughall three constructs had generally similar pharmacokinetics whenadministered to a naive animal, they differed quantitatively in theirimmunogenicity. Antibodies were detected more frequently, and titerswere significantly higher (P < 0.05)in both healthy and septic baboons that received the 4.0-domain TNF-bpconstruct, compared with animals receiving the 2.6-domain construct.When the TNF-bp constructs were administered a second time (21 dayslater), the half-lives of the three constructs were significantlyshorter in animals that had an antibody response after the firstinjection. In contrast, all three TNF-bp constructs were equallyeffective at improving outcome, blocking a systemic TNF- response,and attenuating the cytokine responses when administered at a dose of1.0 mg/kg body wt 1 h before a lethal E. coli infusion. The findings suggest that immunogenicityof TNF-bp constructs can be altered by changing the number offunctional domains, without affecting their capacity to neutralizeTNF- and to abrogate TNF-mediated pathology.