Peptidergic innervation of the human dental pulp: neuropeptide Y

The localization and distribution of the Neuropeptide Y (NPY) has been studied with immunofluorescent methods in the human dental pulp. Immunofluorescence for the NPY has been observed in nervous fibers running in medium and big nerves associated in vascular structures, and in single fibers scattered in the pulpar connective or organized in the subdontoblastic plexus.     

In vitro binding of radioiodinated rat follicle-stimulating hormone to the testis of the mallard duck, Anas platyrhynchos l

The binding of 125I-labelled rat FSH to homogenates and frozen sections of mallard duck testis was investigated. The equilibrium association constant (Ka) in the homogenates (8.5 x 10(9) M-1) was similar to those reported in other avian and mammalian species. Autoradiography suggested that the binding sites for the labelled hormone were localized in the tubular compartment.     

The B subunit of the DNA polymerase alpha-primase complex in Saccharomyces cerevisiae executes an essential function at the initial stage of DNA replication.

The four-subunit DNA polymerase alpha-primase complex is unique in its ability to synthesize DNA chains de novo, and some in vitro data suggest its involvement in initiation and elongation of chromosomal DNA replication, although direct in vivo evidence for a role in the initiation reaction is still lacking. The function of the B subunit of the complex is unknown, but the Saccharomyces cerevisiae POL12 gene, which encodes this protein, is essential for cell viability. We have produced different pol12 alleles by in vitro mutagenesis of the cloned gene. The in vivo analysis of our 18 pol12 alleles indicates that the conserved carboxy-terminal two-thirds of the protein contains regions that are essential for cell viability, while the more divergent NH2-terminal portion is partially dispensable. The characterization of the temperature-sensitive pol12-T9 mutant allele demonstrates that the B subunit is required for in vivo DNA synthesis and correct progression through S phase. Moreover, reciprocal shift experiments indicate that the POL12 gene product plays an essential role at the early stage of chromosomal DNA replication, before the hydroxyurea-sensitive step. A model for the role of the B subunit in initiation of DNA replication at an origin is presented.     

Inhibition of the mTOR pathway and reprogramming of protein synthesis by MDM4 reduce ovarian cancer metastatic properties

Epithelial ovarian cancer (EOC) is a highly heterogeneous disease with a high death rate mainly due to the metastatic spread. The expression of MDM4, a well-known p53-inhibitor, is positively associated with chemotherapy response and overall survival (OS) in EOC. However, the basis of this association remains elusive. We show that in vivo MDM4 reduces intraperitoneal dissemination of EOC cells, independently of p53 and an immune-competent background. By 2D and 3D assays, MDM4 impairs the early steps of the metastatic process. A 3D-bioprinting system, ad hoc developed by co-culturing EOC spheroids and endothelial cells, showed reduced dissemination and intravasation into vessel-like structures of MDM4-expressing cells. Consistent with these data, high MDM4 levels protect mice from ovarian cancer-related death and, importantly, correlate with increased 15 y OS probability in large data set analysis of 1656 patients. Proteomic analysis of EOC 3D-spheroids revealed decreased protein synthesis and mTOR signaling, upon MDM4 expression. Accordingly, MDM4 does not further inhibit cell migration when its activity towards mTOR is blocked by genetic or pharmacological approaches. Importantly, high levels of MDM4 reduced the efficacy of mTOR inhibitors in constraining cell migration. Overall, these data demonstrate that MDM4 impairs EOC metastatic process by inhibiting mTOR activity and suggest the usefulness of MDM4 assessment for the tailored application of mTOR-targeted therapy.Subject terms: Metastasis, Mechanisms of disease     

Cardiovascular pathology possibly associated with ketamine/xylazine anesthesia in Dutch belted rabbits.

BACKGROUND AND PURPOSE: After myocardial necrosis and fibrosis was observed in five rabbits which had been anesthetized a variable number of times, the potential relationship of these lesions and anesthesia was evaluated in 35 other rabbits. METHODS: Anesthesia was induced by intramuscular administration of ketamine and xylazine followed by infusion of lactated Ringer's solution also containing ketamine and xylazine. Group A rabbits (n = 9) were subjected to multiple anesthesias and were evaluated by echocardiography, thoracic radiography, electrocardiography, determination of serum coronavirus titer, vitamin E concentration, and complete necropsy. Prior to a single acute procedure followed by necropsy, group B rabbits (n = 11) were evaluated by echocardiography only. Group C rabbits (n = 10) had never been anesthetized and were necropsied after euthanasia. Group D rabbits (n = 5) had intermediate anesthesia exposure history and were evaluated by echocardiography only. Myocardial fibrosis was scored semi-quantitatively on a scale of 0 to 4. RESULTS: Canine coronavirus test results were negative; hypovitaminosis E was evident, and fibrosis scores were significantly increased in group A, compared with group B or group C, rabbits. CONCLUSION: Etiologic differentials included alpha2-agonist-mediated coronary vasoconstriction with associated myocardial hypoperfusion, hypovitaminosis E and free radical injury, and other anesthetic-induced physiologic trespass.     

The NOP receptor involvement in both withdrawal- and CCk-8-induced contracture responses of guinea pig isolated ileum after acute activation of κ-opioid receptor

In isolated guinea-pig ileum (GPI), the κ-opioid acute withdrawal response is under the control of several neuronal signaling systems, including the μ-opioid, the A₁-adenosine and the CB₁ receptors, which are involved in the inhibitory control of the κ-withdrawal response. After κ-opioid system stimulation, indirect activation of μ-opioid, A₁-adenosine and CB₁ systems is prevented by the peptide cholecystokinin-8 (CCk-8). In the present study, we have investigated whether the NOP system is also involved in the regulation of the acute κ-withdrawal response. Interestingly, we found that in GPI preparation, the NOP system is not indirectly activated by the κ-opioid receptor stimulation, but instead this system is able by itself to directly regulate the acute κ-withdrawal response. Specifically, our results clearly highlight first the existence of an endogenous tone of the NOP system in GPI, and second that it behaves as a functional anti-opioid system. We also found that, the NOP receptor system is involved in the regulation of the CCk-8-induced contracture intensity, only when in the presence of the κ-opioid receptor stimulation. This effect seems to be regulated by an activation threshold mechanism. In conclusion, the NOP system could act as neuromodulatory system, whose action is strictly related to the modulation of both excitatory and inhibitory neurotransmitters released in GPI enteric nervous system.