Nitric oxide synthase-dependent NADPH-diaphorase activity in the optic lobes of male and female Ceratitis capitata mutants

Nitric oxide (NO) is acknowledged as a messenger molecule in the nervous system with a pivotal role in the modulation of the chemosensory information. It has been shown to be present in the optic lobes of several insect species. In the present study, we used males and females from four different strains of the medfly Ceratitis capitata (Diptera, Tephritidae): or; or,wp (both orange eyed); w,M360 and w,Heraklion (both white eyed), as models to further clarify the involvement of NO in the mutants' visual system and differences in its activity and localization in the sexes. Comparison of the localization pattern of NO synthase (NOS), through NADPH-diaphorase (NADPHd) staining, in the optic lobes of the four strains, revealed a stronger reaction intensity in the retina and in the neuropile region lamina than in medulla and lobula. Interestingly, the intensity of NADPHd staining differs, at least in some strains, in the optic lobes of the two sexes; all the areas are generally strongly labelled in the males of the or and w,M360 strains, whereas the w,Heraklion and or,wp mutants do not show evident sex-dependent NADPHd staining. Taken as a whole, our data point to NO as a likely transmitter candidate in the visual information processes in insects, with a possible correlation among NOS distribution, eye pigmentation and visual function in C. capitata males. Moreover, NO could influence behavioural differences linked to vision in the two sexes.     

Insulin prolongs the QTc interval in humans

Insulin hyperpolarizes plasma membranes; we tested whether insulin affects ventricular repolarization. In 35 healthy volunteers, we measured the Q-T interval during electrocardiographic monitoring in the resting state and in response to hyperinsulinemia (euglycemic 1-mU. min(-1). kg(-1) insulin clamp). A computerized algorithm was used to identify T waves; Bazett's formula was employed to correct Q-T (QTc) by heart rate (HR). In the resting state, QTc was inversely related to indexes of body size (e.g., body surface area, r = -0.53, P = 0.001) but not to indexes of body fatness. During the clamp, HR (67 +/- 1 to 71 +/- 1 beats/min, P < 0.0001) and plasma norepinephrine levels (161 +/- 12 to 184 +/- 10 pg/ml, P < 0.001) increased. QTc rose promptly and consistently, averaging 428 +/- 6 ms between 30 and 100 min (P = 0.014 vs. the resting value of 420 +/- 5 ms). Fasting serum potassium (3.76 +/- 0.03 mM) declined to 3. 44 +/- 0.03 mM during insulin. After adjustment for body size, resting QTc was directly related to fasting plasma insulin (partial r = 0.43, P = 0.01); furthermore, QTc was inversely related to serum potassium levels both in the fasting state (partial r = -0.16, P < 0. 04) and during insulin stimulation (partial r = -0.47, P = 0.003). Neither resting nor clamp-induced QTc was related to insulin sensitivity. Physiological hyperinsulinemia acutely prolongs ventricular repolarization independent of insulin sensitivity. Both insulin-induced hypokalemia and adrenergic activation contribute to this effect.     

The Ca(2+)-activated K(+) channel KCa3.1 compartmentalizes in the immunological synapse of human T lymphocytes

T cell receptor engagement results in the reorganization of intracellular and membrane proteins at the T cell-antigen presenting cell interface forming the immunological synapse (IS), an event required for Ca²⁺ influx. KCa3.1 channels modulate Ca²⁺ signaling in activated T cells by regulating the membrane potential. Nothing is known regarding KCa3.1 membrane distribution during T cell activation. Herein, we determined whether KCa3.1 translocates to the IS in human T cells using YFP-tagged KCa3.1 channels. These channels showed electrophysiological and pharmacological properties identical to wild-type channels. IS formation was induced by either anti-CD3/CD28 antibody-coated beads for fixed microscopy experiments or Epstein-Barr virus-infected B cells for fixed and live cell microscopy. In fixed microscopy experiments, T cells were also immunolabeled for F-actin or CD3, which served as IS formation markers. The distribution of KCa3.1 was determined with confocal and fluorescence microscopy. We found that, upon T cell activation, KCa3.1 channels localize with F-actin and CD3 to the IS but remain evenly distributed on the cell membrane when no stimulus is provided. Detailed imaging experiments indicated that KCa3.1 channels are recruited in the IS shortly after antigen presentation and are maintained there for at least 15–30 min. Interestingly, pretreatment of activated T cells with the specific KCa3.1 blocker TRAM-34 blocked Ca²⁺ influx, but channel redistribution to the IS was not prevented. These results indicate that KCa3.1 channels are a part of the signaling complex that forms at the IS upon antigen presentation. T cell activation; ion channels; membrane distribution     

New observations and reinterpretation on the enigmatic taxon Colombitherium (?Pyrotheria,Mammalia) from Colombia

Abstract: The controversial taxon Colombitherium tolimense (Mammalia) (probably Late Eocene in age) from Colombia, although known for nearly 40 years, still bears much mystery. Aside from the problematic ordinal attribution of the holotype and only specimen, its determination as an upper or lower jaw remains a highly debated issue. New observations include the presence of a contact facet on the distal face of the most posterior tooth, which indicates that the fragmentary jaw preserves three premolars and two molars; the M3, unpreserved but present, being most probably reduced. This new interpretation completely fits the morphology of the teeth. Furthermore, the shape of these latter and the deeper wear encompassed by their lingual part relative to the labial one is typical of upper dentition. This is in agreement with the internal curving of the roots of the anterior premolars and with several other arguments that lead interpreting the holotype of C. tolimense as a maxillary bearing P2‐M2. This new interpretation deepens the morphological gap between Colombitherium and other pyrotherians (except Proticia) and challenges further its referral to Pyrotheria. The peculiar morphology of Colombitherium relative to other pyrotherians is indeed striking. In fact, Colombitherium has nothing in common with pyrotherians but bilophodont cheek teeth, a feature largely widespread in placental mammals. It is here referred to ?Pyrotheria until additional evidence of its relationships is known. Associated with the putative removal of Proticia from Pyrotheria as argued by some authors, the hypothetical removal of Colombitherium from the order would adjust the widely accepted assumption that the pyrotherian bilophodont cheek teeth originated from bunodont cheek teeth. It would also make an origin from lophodont forms plausible. This in turn would have critical relevance, especially to the hypothesis that pyrotherians are notoungulates.