Increase of thyrotropin-releasing hormone immunoreactivity in the nucleus of the solitary tract following bilateral lesions of the hypothalamic paraventricular nuclei

The effects of bilateral electrolytic lesions of hypothalamic paraventricular nucleus on thyrotropin-releasing hormone (TRH)-immunoreactive fibers of the nucleus of the solitary tract were studied by both immunocytochemistry and radioimmunoassay. Contrasting with a near disappearance of TRH immunoreactivity in the median eminence, both morphological and biochemical approaches demonstrate that such hypothalamic lesions induced significant increase of TRH immunoreactivity in the nucleus of solitary tract. These results confirm that TRH fibers of the nucleus of the solitary tract do not originate in the hypothalamic paraventricular nucleus (PVN). They further indicate that these TRH neurons projecting to the nucleus of the solitary tract are strongly influenced by neurons located within the PVN area.     

Hexachlorobenzene, a dioxin-like compound, disrupts auditory function in rat

Hexachlorobenzene (HCB) is a dioxin-like compound widely distributed in the environment. In this study, we investigated the effects of HCB on the cochlea. Conscious free-moving rats were given HCB per os daily for 4 weeks at doses of 0.16, 4 or 16 mg/kg in olive oil, whereas the control group received olive oil only. The effects of HCB were evaluated at various time intervals, by measuring auditory nerve acoustic thresholds and plasma thyroid hormone concentration by radioimmunoassay. Histological evaluation involved surface preparation and scanning electron microscopy observations of cochlear hair cells. At a dose of 0.16 mg/kg, HCB induced no loss of acoustic sensitivity, whereas at 4 mg/kg, it induced cochlear sensitivity deficits at the mid-frequencies (2-16 kHz) with complete recovery once treatment was stopped. At a dose of 16 mg/kg, permanent threshold shifts were observed at all frequencies tested (from 1 to 32 kHz). Morphological studies showed no cochlear hair cell loss or alteration of stereocilia. HCB treatment reduced circulating thyroxine concentrations. Thyroidectomy had no effect on cochlear sensitivity in control animals. Thus, HCB is a potent oto-toxicant, and its ototoxicity may be independent of its thyroidal effects.     

Immunocytochemical ultrastructural study of hypothalamic neurons containing corticotropin-releasing factor in normal and adrenalectomized rats

The neurons of the rat hypothalamus which secrete corticotropin-releasing factor were studied by using a pre-embedding immunocytochemical staining technique that improves both the penetration of immunoreagents within the tissue and the preservation of the ultrastructural morphology of labeled structures. Comparison was made between the subcellular location of corticotropin-releasing factor-41 in perikarya of the paraventricular nucleus and axons of the median eminence, both in intact and adrenalectomized animals either untreated or 24 h after the intracerebral injection of colchicine. Morphometric analysis of the numerical density and of the diameter of corticotropin-releasing factor immunoreactive neurosecretory granules in axons of the median eminence of rats not treated with colchicine, indicated that the main modifications induced by adrenalectomy concerned (1) the differential repartition of labeled granules within the preterminal and terminal axonal portions of the median eminence, and (2) the enlargement of the diameter of labeled granules contained in these axons (from 98 nm to 165 nm). In the hypothalamus of intact and adrenalectomized rats, colchicine treatment increased the number of corticotropin-releasing factor-immunoreactive granules in the neuronal perikarya and reduced their number in the axons, but both these variations were much more marked in adrenalectomized rats. Although the corticotropin-releasing factor immunoreactive granules that accumulated in the perikarya after colchicine treatment were slightly smaller than those in the corresponding axons, the diameter of perikaryal-labeled granules was larger in adrenalectomized than in intact animals (129 nm vs 93 nm). These findings fit the idea that adrenalectomy markedly stimulates both the synthesis and axonal excretion of secretory granules in the hypothalamic neurons secreting corticotropin-releasing factor. They also indicate that suppression of circulating corticosteroids induces qualitative modifications in these neurons leading to the visualization of larger neurosecretory granules, which may reflect differential synthesis and granular packing of synergistic peptides other than corticotropin-releasing factor and/or changes in the process of intragranular maturation of hormonal material.     

Comparative immunocytochemical study of the catecholaminergic and peptidergic afferent innervation to the dorsal vagal complex in rat and guinea pig

Light and electron microscopic immunocytochemistry was used to study the fine structural organization of the catecholaminergic and hypothalamic peptidergic innervation of the dorsal vagal complex of the medulla oblongata in the rat and guinea pig, the latter of which is known to lack central adrenergic neurons. In the rat, adrenergic fibers immunoreactive to phenylethanolamine-N-methyltransferase were concentrated in the dorsal motor nucleus of the vagus, where they established frequent symmetric synapses with dendrites and perikarya. On the other hand, the density of both oxytocin- and corticotropin-immunoreactive fibers appeared far lower in this nucleus than in the dorsal regions of the nucleus of the tractus solitarius, where they formed asymmetric synapses with small dendrites. In tissue treated for the dual labeling of two neuronal antigens, oxytocin- or corticotropin-reactive fibers were in close contact with adrenergic neurons in this dorsal medullary region. In the guinea pig, unlike the rat, the dorsal motor nucleus of the vagus contained large amounts of oxytocin- and corticotropin-reactive fibers, which formed many symmetric synapses with perikarya and dendrites. Taken together, these data suggest that the control of vagal preganglionic neurons by hypothalamic peptidergic neurons involves a bisynaptic neuronal pathway including adrenergic medullary neurons in the rat, whereas it is direct in the guinea pig, which lacks this adrenergic relay.     

Basal insulin dose in 40 type 1 diabetic patients remains stable 1 year after educational training in flexible insulin therapy

AimBasal insulin dose (BID) determination is the key to successful flexible insulin therapy (FIT). As our hypothesis was that BID changes over time, the primary objective of the present study was to determine the changes in BID 1 year after a therapeutic educational programme on FIT.MethodsThis single-centre retrospective study recruited the first 40 type 1 adult diabetic patients undergoing an educational FIT programme, which was conducted over a 4-day hospital stay and included a carbohydrate-fasting test.ResultsPatients’ BIDs decreased between Day 0 and Day 4 after the programme (0.31 ± 0.11 IU/kg/day vs 0.27 ± 0.09 IU/kg/day; P < 0.0001), and was increased at 1 year (0.29 ± 0.09 IU/kg/day; P = 0.004). There was no significant variation in prandial insulin requirements. A tendency toward a reduction in HbA_1c was observed at 1 year (8.3 ± 1.4% vs 8.1 ± 1.6%; P = 0.075), with a decrease by more than 0.5% in 37.5% of patients. Body weight increased at 1 year (66.9 ± 10.4 kg vs 68.1 ± 10.7 kg; P = 0.003), and the gain was greater than 5% in 7.5% of patients. Frequency of mild hypoglycaemia either remained stable (40%) or decreased (30%). Only nine patients (baseline HbA_1c 8.03 ± 1.7%, baseline BID 0.27 ± 0.09 IU/kg/day) had BID increases more than 20%, with no changes in prandial insulin requirements and no distinctive phenotype. Baseline HbA_1c, and BID have an impact on the BID at 1 year of approximately 0.3 IU/kg/day in most patients.ConclusionThe stability of BID over 1 year, with values close to 0.3 IU/kg/day associated with a trend towards improvement in HbA_1c, reduction in the frequency of mild hypoglycaemic episodes and absence of major weight gain, supports the relevance of FIT educational training.     

Block of neural Kv1.1 potassium channels for neuroinflammatory disease therapy

OBJECTIVE: We asked whether blockade of voltage-gated K+ channel Kv1.1, whose altered axonal localization during myelin insult and remyelination may disturb nerve conduction, treats experimental autoimmune encephalomyelitis (EAE). METHODS: Electrophysiological, cell proliferation, cytokine secretion, immunohistochemical, clinical, brain magnetic resonance imaging, and spectroscopy studies assessed the effects of a selective blocker of Kv1.1, BgK-F6A, on neurons and immune cells in vitro and on EAE-induced neurological deficits and brain lesions in Lewis rats. RESULTS: BgK-F6A increased the frequency of miniature excitatory postsynaptic currents in neurons and did not affect T-cell activation. EAE was characterized by ventriculomegaly, decreased apparent diffusion coefficient, and decreased (phosphocreatine + beta-adenosine triphosphate)/inorganic phosphate ratio. Reduced apparent diffusion coefficient and impaired energy metabolism indicate astrocytic edema. Intracerebroventricularly BgK-F6A-treated rats showed attenuated clinical EAE with unexpectedly reduced ventriculomegaly and preserved apparent diffusion coefficient values and (phosphocreatine + beta-adenosine triphosphate)/inorganic phosphate ratio. Thus, under BgK-F6A treatment, brain damage was dramatically reduced and energy metabolism maintained. INTERPRETATION: Kv1.1 blockade may target neurons and astrocytes, and modulate neuronal activity and neural cell volume, which may partly account for the attenuation of the neurological deficits. We propose that Kv1.1 blockade has a broad therapeutic potential in neuroinflammatory diseases (multiple sclerosis, stroke, and trauma).     

Effects of pinealectomy on glucagon responsiveness to hypoglycaemia induced by insulin injections in fed rats

The glucagon response to insulin-induced hypoglycaemia was tested in rats that had been pinealectomised (Px), pinealectomised and fitted with melatonin implants (Px + MLT), or sham-operated (SO). The glucagon response to hypoglycaemia observed in SO rats (216 +/- 27 pg x ml(-1) at baseline versus 397 +/- 35 pg x ml(-1) at the hypoglycaemic peak, mean +/- S.D.) was stronger than that in Px rats (180 +/- 37 pg x ml(-1) and 229 +/- 21 pg x ml(-1), respectively) and weaker than that in Px + MLT rats (256 +/- 19 pg x ml(-1) and 516 +/- 11 pg x ml(-1), respectively). Our data indicate that the capacity to release glucagon during insulin-induced hypoglycaemia is altered in pinealectomised rats.     

Suppression of the DNA repair defects of BRCA2-deficient cells with heterologous protein fusions

The BRCA2 tumor suppressor plays an important role in the repair of DNA damage by homologous recombination, also termed homology-directed repair (HDR). Human BRCA2 is 3,418 aa and is composed of several domains. The central part of the protein contains multiple copies of a motif that binds the Rad51 recombinase (the BRC repeat), and the C terminus contains domains that have structural similarity to domains in the ssDNA-binding protein replication protein A (RPA). To gain insight into the role of BRCA2 in the repair of DNA damage, we fused a single (BRC3, BRC4) or multiple BRC motifs to the large RPA subunit. Expression of any of these protein fusions in Brca2 mutant cells substantially improved HDR while suppressing mutagenic repair. A fusion containing a Rad52 ssDNA-binding domain also was active in HDR. Mutations that reduced ssDNA or Rad51 binding impaired the ability of the fusion proteins to function in HDR. The high level of spontaneous chromosomal aberrations in Brca2 mutant cells was largely suppressed by the BRC-RPA fusion proteins, supporting the notion that the primary role of BRCA2 in maintaining genomic integrity is in HDR, specifically to deliver Rad51 to ssDNA. The fusion proteins also restored Rad51 focus formation and cellular survival in response to DNA damaging agents. Because as little as 2% of BRCA2 fused to RPA is sufficient to suppress cellular defects found in Brca2-mutant mammalian cells, these results provide insight into the recently discovered diversity of BRCA2 domain structures in different organisms.     

An investigation of serotonergic involvement in the regulation of ACTH and corticosterone in the olfactory bulbectomized rat.

The bilateral olfactory bulbectomy resulted in significantly higher plasma concentration of corticosterone, but not of ACTH in basal conditions and much higher plasma ACTH and corticosterone concentrations after 15 min of immobilization stress than were observed in sham-operated animals. Daily treatment with fluoxetine-a specific serotonin reuptake inhibitor-(15 mg/kg/day) had no effect on basal ACTH and corticosterone concentrations in OB rats. Fluoxetine treatment caused lower levels of ACTH, but not of corticosterone secretion, in response to immobilization stress. Bulbectomy significantly reducing 5-HT concentration in the amygdala. Stress increased serotonergic activity in the hypothalamus but not in the amygdala of OB rats. Chronic fluoxetine treatment of both unstressed and stressed OB rats resulted in a lower turnover rate in the two structures. Our results suggest that the hypercorticosteronemia observed after bulbectomy in unstressed OB rats is independent of the serotonergic system in both hypothalamus and amygdala. In contrast, they also demonstrate hypothalamic 5-HT changes in the HPA hyperactivity of OB rats in response to stress. Chronic fluoxetine treatment may normalize pituitary ACTH secretion in response to stress, possibly desensitization of the 5-HT receptors in the hypothalamus due to 5-HT being move available at the synapses.