(R)-citramalate synthase in methanogenic archaea

The Methanococcus jannaschii gene MJ1392 was cloned, and its protein product was hyperexpressed in Escherichia coli. The resulting protein was purified and shown to catalyze the condensation of pyruvate and acetyl coenzyme A, with the formation of (R)-citramalate. Thus, this gene (cimA) encodes an (R)-citramalate synthase (CimA). This is the first identification of this enzyme, which is likely involved in the biosynthesis of isoleucine.     

The sec6/8 complex is located at neurite outgrowth and axonal synapse-assembly domains

The molecules that specify domains on the neuronal plasma membrane for the delivery and accumulation of vesicles during neurite outgrowth and synapse formation are unknown. We investigated the role of the sec6/8 complex, a set of proteins that specifies vesicle targeting sites in yeast and epithelial cells, in neuronal membrane trafficking. This complex was found in layers of developing rat brain undergoing synaptogenesis. In cultured hippocampal neurons, the sec6/8 complex was present in regions of ongoing membrane addition: the tips of growing neurites, filopodia, and growth cones. In young axons, the sec6/8 complex was also confined to periodic domains of the plasma membrane. The distribution of synaptotagmin, synapsin1, sec6, and FM1-43 labeling in cultured neurons suggested that the plasma membrane localization of the sec6/8 complex preceded the arrival of synaptic markers and was downregulated in mature synapses. We propose that the sec6/8 complex specifies sites for targeting vesicles at domains of neurite outgrowth and potential active zones during synaptogenesis.     

Effects of Growth Temperature on Some Structural Properties of Crystalline Lamellae in Starches Extracted from Sweet Potatoes (Sunnyred and Ayamurasaki)

Thermodynamic characteristics defining gelatinisation (modelled as ‘melting’) processes for starches extracted from two sweet potato cultivars (Ayamurasaki and Sunnyred) grown in soil at different temperatures (15, 21, 27 or 33 °C) were studied using high sensitivity differential scanning calorimetry (DSC). The gelatinisation temperatures for all starches were elevated significantly with increasing growth temperature. A linear correlation between growth temperature and gelatinisation temperature of extracted starches was found for both cultivars. The increase of gelatinisation temperatures was associated with almost constant or slightly increased enthalpy (Ayamurasaki) but with moderate elevation of crystalline lamellae thickness (Sunnyred). Elevation of gelatinisation temperatures by 8‐9 °C by DSC for starches heated in 1.5 M aqueous KCl were found in comparison with aqueous starch dispersions. The crystalline polymorphic form was confirmed as A‐type using wide angle X‐ray scattering (WAXS). The values of the thermodynamic surface parameters characterising the faces of crystalline lamellae of these starches were calculated. Two endothermic peaks were observed by DSC for starches grown at 15 °C when suspended in 1.5 M KCl solution. The first peak was ascribed to the melting of B‐type structure while the second one was attributed to the melting of A‐type structure. As a conclusion it was suggested that these starches contain C‐type polymorphic structures.     

Fish inner ear otolith size and bilateral asymmetry during development

Cholecystokinin (CCK) is one of the first discovered gastrointestinal hormones and one of the most abundant neuropeptides in the brain. Two types of CCK receptors have been identified: (1) CCK-A receptors are mainly located in the periphery, but are also found in some areas of the CNS; and (2) CCK-B receptors are widely distributed in the brain. Major biological actions of CCK are the reduction of food intake and the induction of anxiety-related behavior. Inhibition of feeding is mainly mediated by the A-type receptors, whereas anxiety-like behavior is induced by stimulating B-type receptors. This paper presents new findings on the effects of the biologically active CCK agonists, CCK-8S, CCK-4, and A71378. The results reviewed suggest that the hypophagic effects of CCK are strongly dependent on the experimental design, sex, and age of the rats. For example, food intake measured during the night or after food deprivation is reduced by CCK-8S in young adult and aged rats, whereas, under fixed feeding conditions, CCK-8S does not inhibit food intake in young adult rats. The sensitivity to the hypophagic CCK effect increases with age in male and female rats; however, female rats are less sensitive to the CCK action. Further, using a nongenetic and non-stressful model of obesity due to unspecific postnatal overfeeding, the satiating effect of moderate CCK-8S doses is weaker in obese than in normal rats. Again, the hypophagic effect is more pronounced in male than in female obese and normal rats. Considering that aversive reactions in rats are markedly influenced by strain and breeding-line variations, research results in this area are critically reviewed. It is shown that anxiety-like symptoms can only be induced by a selectively acting CCK-B agonist, whereas mixed CCK-A and -B agonists and selective CCK-A agonists fail to change behavior in anxiety tests. CCK-4 induces stable and reproducible anxiogenic-like behavior only in certain rat strains. Moreover, CCK-4 effects can be demonstrated in the conflict test, in the ultrasonic vocalization test in rat pups, on the elevated plus maze, and in the black and white box, but not in the social interaction test. CCK has also been reported to modulate memory processes. On the one hand, CCK-8S and CCK-4 enhanced habituation to the novelty of a hole board. On the other hand, repeated administration of CCK-8S did not improve maze performance in aged rats. The literature on the behavioral pharmacology of CCK is rife with inconsistency and contradiction. The major biological actions of CCK depend on the receptor selectivity of the CCK fragments used and on organismic and procedural variables. All these variables potentially influence behavioral responses in rats. Therefore, in CCK research more attention should be paid to the importance of these methodological factors.     

Conceptual behavior in pigeons: Categories, subcategories, and pseudocategories.

In 3 experiments with a total of 16 Ss, we explored how pigeons learn to classify diverse pictures of cats, flowers, cars, and chairs and later how they accurately categorize brand-new pictures from these classes. Using a 4-key forced-choice procedure, Ss in Exp 1 discriminated individual examples within each of the categories from one another (subcategory training); nevertheless, errors were disproportionately conceptual in nature, with Ss more likely to confuse examples within a given category than between different categories. Ss in Exp 2 trained to classify pictures into human language categories (category training) learned far faster and more completely than Ss trained to sort the same pictures into totally arbitrary groupings (pseudocategory training). Finally, in Exp 3, category-trained and subcategory-trained Ss were tested on normally oriented pictures, on left–right reversals, and on top–bottom reversals. Subcategory-trained Ss responded less accurately on both kinds of reversed pictures and less accurately on top–bottom than on left–right reversals; category-trained Ss were less affected by both types of picture reversals, only top–bottom reversals decrementing their performance. Results suggest that many words in our language denote clusters of related visual stimuli, which pigeons also see as highly similar. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Metabotropic glutamate receptor antagonists, like GABA(B) antagonists, potentiate dorsal root-evoked excitatory synaptic transmission at neonatal rat spinal motoneurons in vitro

Recordings of whole-cell synaptic current responses elicited by electrical stimulation of dorsal roots were made from motoneurons, identified by antidromic invasion, in isolated spinal cord preparations from five- to eight-day-old Wistar rats. Supramaximal electrical stimulation of the dorsal root evoked complex excitatory postsynaptic currents with mean latencies (+/- S.E.M.) of 6.1 +/- 0.26 ms, peak amplitude of -650 +/- 47 pA and duration of 4.30 +/- 0.46 s (n=34). All phases of excitatory postsynaptic currents were potentiated to approximately 20% above control levels in the presence of the metabotropic glutamate receptor antagonists S-2-amino-2-methyl-4-phosphonobutanoate (MAP4; 200 microM; n=15) and 2S, 1'S,2'S-2-methyl-2-(carboxycyclopropyl)glycine (MCCG; 200 microM; n=9). A similar level of potentiation was produced by the GABA(B) receptor antagonist 3-N[1-(S)-(3,4-dichlorophenyl)ethyl]amino-2-(S)-hydroxypropyl-P-benzyl-p hosphinic acid (CGP55845; 200 nM; n=5). MAP4 (200 microM) produced a six-fold rightward shift in the concentration-effect plot for the depressant action of the metabotropic glutamate receptor agonist S-2-amino-4-phosphonobutanoate (L-AP4), whereas CGP55845 produced no significant change in the potency of L-AP4. MAP4 did not antagonize the depressant actions of baclofen (n=8), 1S,3S-1-aminocyclopentane-1,3-dicarboxylate (n=4) or 2-S,1'S,2'S-2-(carboxycyclopropyl)glycine (n=4). The metabotropic glutamate receptor antagonists produced no change in the holding current of any of the neurons, indicating that they had no significant postsynaptic excitatory actions. These results are the first to indicate a possible physiological role for metabotropic glutamate receptors in the spinal cord. Like GABA(B) receptors, they control glutamatergic synaptic transmission in the segmental spinal pathway to motoneurons. This is likely to be a presynaptic control mechanism.     

Prestimulus effects on startle magnitude: Sensory or motor?

Startle may be inhibited when the startling event is preceded by a stimulus; this is called prepulse inhibition (PPI) when the prestimulus is weak and nonstartling (s) and paired pulse inhibition when the prestimulus elicits startle (S1). The authors examined the relationship of these measures across species and tested whether paired pulse inhibition-like PPI-is independent of the startling effects of the prestimulus. PPI (s-S1 configuration) and paired pulse inhibition (S1-S2 configuration) were elicited in 1 test, using similar stimulus parameters in rats and humans. The amount of PPI and paired pulse inhibition was significantly correlated within subjects in both rats and humans. Paired pulse inhibition was not diminished when the startling effects of S1 were eliminated by a weak prepulse (s-S1-S2 configuration), nor was it enhanced when these prepulse effects were eliminated by the dopamine agonist apomorphine (in rats). Despite apparent differences in the inhibitory processes mediating PPI and paired pulse inhibition, both are independent of the motoric response to the prestimulus. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Quantitative determination of thebaine in Papaver bracteatum by high-pressure liquid chromatography

A method is described for the quantitative analysis of thebaine from Papaver bracteatum, using a single high-pressure liquid chromatographic column. The procedure gives base-line separation of thebaine without the need for gradient elution equipment, and can be completed within 12 min. Thebaine isolated by this technique was shown to be pure, regardless of the age of plant or plant part from which it was obtained.     

A prospective, randomized study of the use of platelet concentrates irradiated with ultraviolet-B light in patients with hematologic malignancy

BACKGROUND: Irradiation of platelet concentrates (PCs) with ultraviolet-B (UVB) light inactivates the contaminating white cells and might be an alternative to filtration for the prevention of alloimmunization to HLA antigens and subsequent refractoriness to further platelet transfusions in multiply transfused patients with bone marrow failure. STUDY DESIGN AND METHODS: Patients with hematologic malignancy, mainly acute myeloid leukemia, were prospectively assigned in a random manner to receive either UVB-irradiated or control, nonirradiated PCs. All patients were given red cells that were white cell reduced by filtration. Transfusion efficacy and alloimmunization were assessed by means of corrected count increments, requirement for red cells and PCs, and measurement of lymphocyte-reactive antibodies. RESULTS: UVB-irradiated PCs had a clinical efficacy similar to controls as judged by corrected count increments at 1 to 6 and 12 to 24 hours and by the median requirement for red cell and platelet transfusions. Alloimmunization determined by measurements of lymphocyte-reactive antibodies using both conventional and antiglobulin-augmented lymphocytotoxicity techniques was not abolished in recipients of UVB-irradiated PCs (4/30, 13%) but was less than that in controls (5/20, 25%; p = NS). The mean number of platelet transfusion episodes prior to the occurrence of alloimmunization was greater in the control group (27 vs. 10; p = 0.017). CONCLUSION: In this trial, UVB irradiation did not diminish the clinical efficacy of platelet transfusions. There was a small but nonsignificant reduction alloimmunization, but no difference in refractoriness of the two groups was observed. Larger prospective randomized studies are required to confirm these findings and to compare UVB irradiation with white cell reduction.