Molecular architecture of the multiprotein splicing factor SF3b

The splicing factor SF3b is a multiprotein complex essential for the accurate excision of introns from pre-messenger RNA. As an integral component of the U2 small nuclear ribonucleoprotein (snRNP) and the U11/U12 di-snRNP, SF3b is involved in the recognition of the pre-messenger RNA's branch site within the major and minor spliceosomes. We have determined the three-dimensional structure of the human SF3b complex by single-particle electron cryomicroscopy at a resolution of less than 10 angstroms, allowing identification of protein domains with known structural folds. The best fit of a modeled RNA-recognition motif indicates that the protein p14 is located in the central cavity of the complex. The 22 tandem helical repeats of the protein SF3b155 are located in the outer shell of the complex enclosing p14.     

Small nuclear ribonucleoprotein remodeling during catalytic activation of the spliceosome

Major structural changes occur in the spliceosome during its activation just before catalyzing the splicing of pre-messenger RNAs (pre-mRNAs). Whereas changes in small nuclear RNA (snRNA) conformation are well documented, little is known about remodeling of small nuclear ribonucleoprotein (snRNP) structures during spliceosome activation. Here, human 45S activated spliceosomes and a previously unknown 35S U5 snRNP were isolated by immunoaffinity selection and were characterized by mass spectrometry. Comparison of their protein components with those of other snRNP and spliceosomal complexes revealed a major change in protein composition during spliceosome activation. Our data also suggest that the U5 snRNP is dramatically remodeled at this stage, with the Prp19 complex and other factors tightly associating, possibly in exchange for other U5 proteins, and suggest that after catalysis the remodeled U5 is eventually released from the postsplicing complex as a 35S snRNP particle.     

Stabilization of caffeic acid derivatives in Echinacea purpurea L. glycerin extract

Recent work has shown that enzymatic degradation and oxidation of cichoric acid and other caffeic derivatives occurs in Echinacea preparations. However, very little is known as to the means of stabilizing these phytopreparations. To stabilize the glycerin extract of Echinacea purpurea, we have evaluated the effects of 3 natural antioxidants (citric acid, malic acid, and hibiscus extract) on the stability of the major caffeic acid derivatives (caftaric acid, caffeic acid, cichoric acid, and 2-O-feruloyl-tartaric acid). Chlorogenic acid, which normally occurs in an ethanol extract of E. purpurea, was not present in the glycerin extract. The caffeic acid derivatives, with the exception of 2-O-feruloyl-tartaric acid, were subject to degradation in the control sample. 2-O-Feruloyl-tartaric acid was stable during the whole testing period. All antioxidant treatments greatly improved the stability of caffeic acid derivatives. Stability was dependent upon the concentration of antioxidant added.     

Inclusion of β-1,3/1,6-glucan in the ornamental fish,Jewel tetra (Hyphessobrycon eques), and its effects on growth,blood glucose,and intestinal histology

The effects of β-1,3/1,6-glucan on Hyphessobrycon eques were assessed after 42 days of feeding diets containing 0 (control group given commercial feed), 0.5, 1, or 2 g β-glucan/kg diet. In total, 180 fish, with an initial weight of 0.43?±?0.03 g, were used. There were 15 fish in each of twelve 42-L aquariums, and there were 3 aquariums of fish for each dietary treatment. The fish were fed until apparent satiety. Performance parameters (final weight, total length, standard length, feed intake, survival rate, weight gain, feed conversion, specific growth rate, and condition factor) and plasma glucose concentration were measured. Histological analysis of the proximal portion of the intestine (width and height of the villi, depth of the crypts, height of the enterocytes, thickness of the muscle layer, and number of goblet cells) was performed. Different levels of the additive did not influence fish performance (for example, final weight: control: 0.63 g, 0.5: 0.60, 1: 0.58, and 2: 0.61). Likewise, there was no influence on the plasma glucose concentration (control: 81.80 mg/dL, 0.5: 75.33, 1: 85.00, and 2: 81.00) and intestinal morphometry of the animals. However, the results showed that 2.0 g/kg of β-1,3/1,6-glucan provided a greater abundance of goblet cells secreting acidic and neutral mucus present in the epithelium (periodic acid-Schiff: 66.67 cells, Alcian blue pH 1.0: 72,67 cells, and Alcian blue pH 2.5: 95.00 cells), showing significant differences when compared to animals in the control group, which may represent better protection of the intestinal epithelium of H. eques.

     

Effect of Glutamate Accumulation During Sourdough Fermentation with Lactobacillus reuteri on the Taste of Bread and Sodium‐Reduced Bread

NaCl is an important contributor to the taste and texture of bread; therefore, it is challenging to reduce NaCl in bread without compromising quality. This study investigated sensory properties of bread with sourdough fermented with Lactobacillus reuteri accumulating glutamate or γ‐aminobutyrate (GABA). Sourdough was fermented with the GABA‐producing L. reuteri 100‐23 and LTH5448 as well as the glutamate‐accumulating L. reuteri 100‐23ΔgadB and TMW1.106. A consumer panel detected significant differences in the taste of bread with 6% addition of sourdough fermented with glutamate‐ or GABA‐producing L. reuteri. Remarkably, this difference was also detected when GABA‐producing L. reuteri 100‐23 was compared with its glutamate‐producing isogenic mutant L. reuteri 100‐23ΔgadB. The intensity of the salty taste of sourdough bread produced with 1% (flour basis) salt was equivalent to the intensity of the salty taste of reference bread produced with 1.5% salt. A trained panel found that sourdough breads (1 or 2% NaCl flour base) had a higher sour and umami taste intensity when compared with reference bread with the same salt content. Bread produced with sourdough fermented with L. reuteri 100‐23ΔgadB consistently had a higher umami taste intensity when compared with other sourdough breads. Neither sourdough addition nor NaCl level influenced bread volume or texture. In conclusion, the use of sourdough fermented with glutamate‐accumulating lactobacilli allowed reduction of NaCl without adverse effects on the taste or other quality attributes of bread.     

Identifying the origins of fishing gear ingested by seabirds: a novel multivariate approach

• 1. The global decline of albatrosses (Diomedidae: Procellariiformes) is thought to have occurred largely as a direct result of fishery‐related mortality. Albatrosses and other large petrels interact with fisheries in several ways, including scavenging used bait and discarded offal, which may contain hooks.
• 2. Hooks that are ingested by breeding birds are often fed to chicks which subsequently regurgitate them shortly before fledging.
• 3. In this study a series of mathematical (cladistic, cluster and principal components) analyses are applied to a sample of 241 items of fishing gear (hook, snood and hook/snood unit) collected from seabird nest sites on Bird Island, South Georgia, and 44 reference gear items provided by four South Atlantic regional fisheries.
• 4. The five separate analyses failed to assign most gear to a particular fishery or to identify any consistent annual trends. The homogeneous nature of the material, which was largely derived from the same manufacturers, meant that gear origin could not be determined. This suggests that hooks found at seabird colonies in this, and potentially other regions, will be of limited use in identifying offending fisheries, unless operators are obliged to deploy gear with unique marks in the future.
• 5. Nevertheless, it is suggested that this approach should work effectively where birds interact with a range of fisheries targeting different species using variable gear. This study therefore represents an innovative approach to the characterization of lost fishing gear with potentially widespread application. Copyright © 2010 John Wiley & Sons, Ltd.

     

Management of airway difficulties during induction of general anesthesia in an American miniature horse with dwarfism

This report describes previously unreported upper airway abnormalities encountered in a 5-month-old American miniature horse colt presented for elective surgery. Caution should be exercised when administering general anesthesia or heavy sedation to individuals of this breed that present with multiple congenital abnormalities.     

Computed tomography or rhinoscopy as the first-line procedure for suspected nasal tumor: A pilot study

There are no evidence-based guidelines as to whether computed tomography (CT) or endoscopy should be selected as the first-line procedure when a nasal tumor is suspected in a dog or a cat and only one examination can be performed. Computed tomography and rhinoscopic features of 17 dogs and 5 cats with a histopathologically or cytologically confirmed nasal tumor were retrospectively reviewed. The level of suspicion for nasal neoplasia after CT and/or rhinoscopy was compared to the definitive diagnosis. Twelve animals underwent CT, 14 underwent rhinoscopy, and 4 both examinations. Of the 12 CT examinations performed, 11 (92%) resulted in the conclusion that a nasal tumor was the most likely diagnosis compared with 9/14 (64%) for rhinoscopies. Computed tomography appeared to be more reliable than rhinoscopy for detecting nasal tumors and should therefore be considered as the first-line procedure.     

Fate of ACE-inhibitory peptides during the bread-making process: Quantification of peptides in sourdough,bread crumb,steamed bread and soda crackers

This study compared the concentration of angiotensin-converting enzyme (ACE) inhibitory peptides at different stages of the bread-making process, including kneading, proofing, and final products. Steamed bread, baked bread, and soda crackers were produced with 3–20% addition of rye malt sourdoughs to assess products differing in their thermal treatment. Eight tripeptides with known or predicted ACE-inhibitory activity were quantified by LC/MS in multiple reaction monitoring (MRM) mode. In wheat sourdough and rye-malt gluten sourdough, IPP was the predominant tripeptide at 58 and 473 μmol kg⁻¹, respectively, followed by LQP, IQP, and LPP. During the bread-making process, peptide concentrations were modified by enzymatic conversions at the dough stage and by thermal reactions during baking. The concentrations of IPP, LPP and VPP remained stable during dough preparation but decreased during thermal treatment; the concentrations of other peptides were changed at the dough stage but remained relatively stable during baking. The cumulative concentration of 8 ACE-inhibitory peptides in steamed bread and bread crumb exceeded 60 μmol kg⁻¹, while soda crackers contained less than 3 μmol kg⁻¹. The peptide levels in bread thus likely meet in vivo active concentrations.