Pharmacological effects of Lu AA21004: a novel multimodal compound for the treatment of major depressive disorder.

1-[2-(2,4-Dimethylphenyl-sulfanyl)-phenyl]-piperazine (Lu AA21004) is a human (h) serotonin (5-HT)(3A) receptor antagonist (K(i) = 3.7 nM), h5-HT(7) receptor antagonist (K(i) = 19 nM), h5-HT(1B) receptor partial agonist (K(i) = 33 nM), h5-HT(1A) receptor agonist (K(i) = 15 nM), and a human 5-HT transporter (SERT) inhibitor (K(i) = 1.6 nM) (J Med Chem 54:3206-3221, 2011). Here, we confirm that Lu AA21004 is a partial h5-HT(1B) receptor agonist [EC(50) = 460 nM, intrinsic activity = 22%] using a whole-cell cAMP-based assay and demonstrate that Lu AA21004 is a rat (r) 5-HT(7) receptor antagonist (K(i) = 200 nM and IC(50) = 2080 nM). In vivo, Lu AA21004 occupies the r5-HT(1B) receptor and rSERT (ED(50) = 3.2 and 0.4 mg/kg, respectively) after subcutaneous administration and is a 5-HT(3) receptor antagonist in the Bezold-Jarisch reflex assay (ED(50) = 0.11 mg/kg s.c.). In rat microdialysis experiments, Lu AA21004 (2.5-10.0 mg/kg s.c.) increased extracellular 5-HT, dopamine, and noradrenaline in the medial prefrontal cortex and ventral hippocampus. Lu AA21004 (5 mg/kg per day for 3 days; minipump subcutaneously), corresponding to 41% rSERT occupancy, significantly increased extracellular 5-HT in the ventral hippocampus. Furthermore, the 5-HT(3) receptor antagonist, ondansetron, potentiated the increase in extracellular levels of 5-HT induced by citalopram. Lu AA21004 has antidepressant- and anxiolytic-like effects in the rat forced swim (Flinders Sensitive Line) and social interaction and conditioned fear tests (minimal effective doses: 7.8, 2.0, and 3.9 mg/kg). In conclusion, Lu AA21004 mediates its pharmacological effects via two pharmacological modalities: SERT inhibition and 5-HT receptor modulation. In vivo, this results in enhanced release of several neurotransmitters and antidepressant- and anxiolytic-like profiles at doses for which targets in addition to the SERT are occupied. The multimodal activity profile of Lu AA21004 is distinct from that of current antidepressants.

Hemisphere-dependent endocannabinoid system activity in prefrontal cortex and hippocampus of the Flinders Sensitive Line rodent model of depression.

Altered endocannabinoid (eCB) signalling is suggested as an important contributor to the pathophysiology of depression. To further elucidate this, we conducted a study using a genetic rat model of depression, the Flinders Sensitive Line (FSL), and their controls, the Flinders Resistant Line (FRL) rats. Plasma, right and left prefrontal cortex, and hippocampus were isolated from FSL and FRL rats. We analyzed each region for the eCB anandamide (AEA) and 2-arachidonoylglycerol (2-AG) levels by liquid chromatography/multiple reaction monitoring (LC/MRM), mRNA and protein levels of the cannabinoid type 1 receptor (CB1R), fatty acid amide hydrolase (FAAH) and monoacyl glycerol lipase (MAGL) by real time qPCR and Western blotting. Content of 2-AG was lower in the left side of the hippocampus and prefrontal cortex in FSL rats compared to FRL rats. Inversely, levels of AEA were higher in right hippocampus than in left hippocampus. In plasma, AEA levels were increased and 2-AG decreased. Cannabinoid receptor 1 (Cnr1), Faah and Magl mRNA levels were prominently decreased in right prefrontal cortex of FSL rats as compared to FRL rats. Protein expression of CB1R and FAAH were decreased in left hippocampus. In summary, our data suggest a decreased eCB signalling in the FSL rats, which could contribute to the depressive-like behaviour. Interestingly, the altered eCB system activity appear to be hemisphere-specific in the limbic regions. Our study support the existing literature and showed altered eCB system activity in this particular animal model of depression.

Biosynthesis, secretion and in vivo isotopic labelling of venom of the Egyptian cobra, Naja haje annulifera.

The venom glands of Elapidae differ from those of the Viperidae by lacking an expanded central lumen; the venom is stored in the tubular lumina as well as inside the cells in densely packed secretion granules. Using isotope tracer techniques, it was found that in the Egyptian cobra (Naja haje annulifera) venom is secreted both from pre-existing and from newly-formed granules. The rate of protein biosynthesis peaks at 4-9 days after venom was extracted (milked) from the glands. Highly labelled toxins (1-10 mCi/mmole protein) were isolated in good yield from the venom of snakes chronically intubated and infused i.p. with (3H)-amino acids. Repeated Fluothane (Halothane) anaesthesias and venom collections had no ill effect on venom yield. The radioactive venom and its component toxins retained full biological potency.

Penetrating abdominal wounds--a prospective trial of conservative treatment based on physical signs.

Between 1980-1987 605 patients with penetrating abdominal wounds were admitted to Victoria Hospital; 105 had gunshot wounds of whom 103 had laparotomies with positive findings. It is concluded that virtually all patients with gunshot wounds of the abdomen require laparotomy. The remaining 500 patients presented with stab wounds of the abdomen and were admitted to the surgical wards; of these 357 had physical signs necessitating immediate laparotomy while the remainder were observed. A further 60 observed patients required a late laparotomy and 8 of these patients developed complications; there was 1 death. The overall unnecessary laparotomy rate was 21% (105 patients) with a morbidity of 5.7%. Ninety-five patients had omentum protruding through the wound; 17 were treated conservatively without sequelae and a further 22 had negative findings at laparotomy. There were no significant differences in the morbidity and mortality rates between the conservatively treated group and those requiring immediate laparotomy. It is apparent that there is a definite place for conservative treatment of penetrating stab wounds of the abdomen even when there is protruding omentum. Cases which need particular care are those with stab wounds in and below the 5th left intercostal space and stab wounds of the back and flank. Penetrating stab wounds of the colon, irrespective of site, can be managed by simple closure in the majority of patients.

The ultrastructure of the human epidermis following ultracentrifugation.

Ultracentrifugation of the human epidermis was undertaken to uncover the forces responsible for the establishment and maintenance of the supranuclear melanin cap in basal cells. The wide spacing and elasticity of the tonofibrils caused a selective trapping of melanosomes in this region. At the same time the experiments produced pictures resembling dedifferentiation of the plasma membrane as in the fetal state, and differentiation of the tonofibrillar mass resembling the granular and horny layers. Once the tonofibrils had retracted and massed together, the order of intracellular sedimentation was: melanosome, chromatin, mitochondrion, tonofibril and ribosome.

Spore ultrastructure in Sporothrix schenckii.

Pathogenic strains of Sporothrix schenkii may show triangular spores, whose angular shape is maintained by a tie-beam effect in the inner cell wall structure. This difference in wall structure lies adjacent to a folded and possibly more active part of the spore cytoplasm. The supposed generation of asci in old cultures was simulated by the death of hyphae which are reinvaded by intrahyphal growth with intrahyphal spore production, while true asci were not seen.

High dietary ascorbic acid levels and oxalate crystallization in soft tissues of baboons.

The baboon (Papio ursinus) was used as a model to investigate the possibility of oxalate crystallization in the soft tissues due to long-term feeding of ascorbic acid in high doses. Microscopical inspection in polarized light did not reveal oxalate crystals in any of the soft tissues obtained from 16 baboons fed very high doses of ascorbic acid for a period of 20 months.

Ploidy, ascus formation and recombination in Torulaspora (Debaryomyces) hansenii.

X-ray inactivation studies on the type strain of Torulaspora hansenii carried out to determine ploidy, provided proof that the species has a haplontic life cycle, a fact which hitherto has only been presumed. Observations on the genesis of the ascus by light microscopy and transmission electron microscopy provide no evidence for, what some earlier workers in this field have presumed to be, heterogamous conjugation between a mother cell and its bud. They do, however, show that asci, bearing obliquely-attached, vestigal, bud-like appendages, arise from some cells to form single, non-abstricting, and frequently, recurving protuberances which enlarge. These could, conceivably, be responsible for the impression that abstricted buds are connected to the mother-cells by bent copulatory tubes. The formation during sporulation of elongated protuberances and the presence of a medial, electron-dense line within the electron-translucent layer of the walls of ascospores fixed with OsO4 preclude the possibility of using these features to differentiate between the genera Torulaspora and Debaryomyces. Furthermore, recombinant studies, which involved the use of auxotrophic mutants, indicated that during sporulation the fusion of independent cells accounted for only 0.03-0.6% of the asci formed. The conclusion was reached that somatogamous autogamy must be the main agency of diploidization and that the species is largely inbreeding.