Rieske business: structure-function of Rieske non-heme oxygenases

Rieske non-heme iron oxygenases (RO) catalyze stereo- and regiospecific reactions. Recently, an explosion of structural information on this class of enzymes has occurred in the literature. ROs are two/three component systems: a reductase component that obtains electrons from NAD(P)H, often a Rieske ferredoxin component that shuttles the electrons and an oxygenase component that performs catalysis. The oxygenase component structures have all shown to be of the alpha3 or alpha3beta3 types. The transfer of electrons happens from the Rieske center to the mononuclear iron of the neighboring subunit via a conserved aspartate, which is shown to be involved in gating electron transport. Molecular oxygen has been shown to bind side-on in naphthalene dioxygenase and a concerted mechanism of oxygen activation and hydroxylation of the ring has been proposed. The orientation of binding of the substrate to the enzyme is hypothesized to control the substrate selectivity and regio-specificity of product formation.     

Neurotensin increases endogenous glutamate release in rat cortical slices

In the present study, the effects of the tridecapeptide neurotensin [NT(1-13)] and its fragments, NT(1-7) and NT(8-13), on endogenous glutamate release from rat cortical slices, were evaluated. NT(1-13) (100-1000 nM) slightly increased spontaneous glutamate release, while it was ineffective at 1 and 10 nM concentrations. Neither the biologically active NT fragment NT(8-13) nor the inactive one NT(1-7) affected basal glutamate release. NT(1-13) (1-1000 nM) enhanced potassium (35 mM)-evoked glutamate release displaying a bell-shaped concentration response curve. In addition NT(8-13) (10 nM) increased K+-evoked-glutamate release similarly to the parent peptide (10 nM), while the biologically inactive fragment NT(1-7) (10-100 nM) was ineffective. The effects of NT(1-13) and NT(8-13) were fully counteracted by the selective neurotensin receptor antagonist SR48692 (100 nM). These findings suggest that NT plays a role in regulating cortical glutamate transmission.     

Evidence for a nucleus accumbens CCK2 receptor regulation of rat ventral pallidal GABA levels: a dual probe microdialysis study

We employed dual probe microdialysis in the nucleus accumbens and ipsilateral ventral pallidum of the halothane anaesthetized rat to investigate the effect of intra-accumbens perfusion with the sulphated octapeptide cholecystokinin (CCK-8S, 10-1000 nM, 60 min) alone and in the presence of the selective CCK1 and CCK2 receptor antagonists L-364,718 (10 and 100 nM) and PD134308 (10 nM), tetrodotoxin (TTX, 1000 nM) and the GABA(A) receptor antagonist bicuculline (1000 nM), on dialysate GABA levels in the ventral pallidum. Intra-accumbens perfusion with the 100 and 1000 nM concentration of CCK-8S was associated with a significant decrease (-16+/-3% and -23+/-3% vs basal, respectively) in ventral pallidum GABA levels. The CCK-8S (1000 nM) induced decrease in ventral pallidal dialysate GABA levels was abolished when PD134308, TTX and bicuculline, but not L-364,718, were included into the perfusion medium of the accumbens probe. The data indicate that nucleus accumbens CCK-8S exerts a CCK2 receptor mediated inhibition of ventral pallidal GABA levels. Furthermore, the TTX and bicuculline sensitivity of this effect suggests that this is possibly mediated via CCK2 receptors probably located on local GABA interneurons.     

Aging, Obesity, and Mortality: Misplaced Concern About Obese Older People?

Although there is widespread agreement that obesity (body mass index [BMI] ≥ 30 kg/m(2)) raises health risks, debate has ensued on whether obese older adults are also at greater risk. This study examines the effect of obesity on mortality for younger and older adults to determine whether the risk of obesity is lessened in later life. Data from a 20-year follow-up of a national sample of adults were used to examine the risk of obesity on mortality (N = 6,767). Cox models reveal that obesity raises mortality risk for adults of all ages, but this relationship is nearly twice as strong for persons younger than 50 years of age. Being slightly overweight in later life is associated with lower mortality risk, but obesity raises mortality risk, especially for ischemic heart disease. Obesity in middle age is a grave public health concern, but obesity in later life also merits attention.     

Retrieval and reuse of pituitary secretory granule proteins

The pituitary contains professional secretory cells, devoting a large fraction of their energy to the synthesis of hormones that are stored for secretion in response to a complex mixture of inputs. Ba2+, a substitute for Ca2+, and phorbol ester, a mimic for diacylglycerol, have a synergistic effect on exocytosis. By using these secretagogues, we developed a paradigm in which phorbol ester potentiation of Ba2+-evoked exocytosis produces a robust secretory response in multiple pituitary cell types. Because cells subjected to this stimulatory paradigm remain healthy despite their greatly reduced hormone content, we used this paradigm to study the fate of granule membrane proteins. We examined the turnover of peptidylglycine alpha-amidating monooxygenase (PAM), a membrane enzyme involved in the final maturation of many peptides, and VAMP2, a vesicle soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE). The stability of recently synthesized PAM was increased by sustained exocytosis. Biotinylation studies established that the appearance of integral membrane PAM at the plasma membrane was stimulated along with hormone secretion. PAM biotinylated on the cell surface undergoes cleavage to yield soluble peptidylglycine-alpha-hydroxylating monooxygenase that can then be secreted in a regulated fashion. Consistent with a kiss-and-run or cavicapture mode of secretion (Taraska, J. W., Perrais, D., Ohara-Imaizumi, M., Nagamatsu, S., and Almers, W. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 2070-2075), biotinylated prolactin was also retained by the cells and later released in response to secretagogues. Thus, pituitary cells can retrieve and reuse components of the machinery involved in the final stages of exocytosis (the SNAREs) as well as soluble and membrane granule proteins.     

Identification of ERβ1 and ERβ2 in human seminoma, in embryonal carcinoma and in their adjacent intratubular germ cell neoplasia

Background  

Estrogens exert a role on germ cell physiology of normal human testis through the mediation of the estrogen receptor (ER) beta subtypes. Epidemiological studies evidenced an increased incidence of testicular germ cell cancer after elevated pre-natal estrogen exposure but the expression of estrogen receptors in these testicular neoplasms has not been well elucidated.     

A comparative ultrastructural analysis of spermatozoa in Pleurodema (Anura,Leptodactylidae, Leiuperinae)

This study describes the spermatozoa of 10 of the 15 species of the Neotropical frog genus Pleurodema through transmission electron microscopy. The diversity of oviposition modes coupled with a recent phylogenetic hypothesis of Pleurodema makes it an interesting group for the study of ultrastructural sperm evolution in relation to fertilization environment and egg‐clutch structure. We found that Pleurodema has an unusual variability in sperm morphology. The more variable structures were the acrosomal complex, the midpiece, and the tail. The acrosomal complex has all the structures commonly reported in hyloid frogs but with different degree of development of the subacrosomal cone. Regarding the midpiece, the variability is given by the presence or absence of the mitochondrial collar. Finally, the tail is the most variable structure, ranging from single (only axoneme) to more complex (presence of paraxonemal rod, cytoplasmic sheath, and undulating membrane), with the absence of the typical axial fiber present in hyloid frogs, also shared with some other genera of Leiuperinae. J. Morphol. 277:957–977, 2016. © 2016 Wiley Periodicals, Inc.     

Plasticity of repetitive DNA in response to metal stress in Bryophytes

Abstract

The relationship between environmental stresses and the genome was investigated by examining the behaviour of repetitive DNA in response to lead or cadmium in two Bryophytes, differing from a physiological and an ecological point of view, namely the aquatic moss L. riparium and the terricolous moss F. hygrometrica. Using different experimental approaches, a direct relationship was shown to exist in these two mosses between the metal-induced stress and repetitive DNA. In fact, in both organisms, metal treatment was accompanied by a selective amplification of some GC-rich repetitive DNA sequences forming peculiar agglomerates inside the nucleus; this amplification is quantitatively proportional to the time of exposure of the plants to the metals and stops upon removal of the metal from the culture medium. Results show that ribosomal DNA sequences are involved in this metal-induced repetitive DNA agglomerate formation, although they are not the only repetitive sequences present within the heterochromatic DNA agglomerates. The plasticity of the genome of the Bryophytes in response to external stimuli, and the fact that repetitive DNA is involved in this plasticity are discussed.