Search

Background Glioma is a highly malignant brain tumor, characterized by the poor prognosis and high recurrence rates. Previous studies have confirmed that miRNA-30c-5p is closely associated with tumor cell biological properties. The present study explored the biological role of miR-30c-5p in human glioma malignant behavior and underlying mechanisms. Methods Levels of miR-30c-5p were detected in glioma tissues and adjacent normal tissues. Two glioma cell lines including U87 and U251 were transfected with miR-30c-5p mimic or inhibitors. Cell proliferation was evaluated by MTT assay and colony formation assay. Cell apoptosis and invasive potential of glioma cells were assessed by flow cytometry and transwell assays, respectively. Luciferase reporter assay was performed to validate the target gene of miR-30c-5p. Results Levels of miR-30c-5p were dramatically decreased in glioma tissues as compared to the adjacent normal tissues. Upregulation of miR-30c-5p significantly suppressed cell growth and colony formation, and induced apoptosis in glioma cells. In contrast, inhibition of miR-30c-5p promoted the proliferation and inhibited apoptosis in tumor cells. Furthermore, miR-30c-5p strongly suppresses the invasion of glioma cells. Western blot showed that Bcl-2 was significantly decreased following treatment with miR-30c-5p mimics and increased after miR-30c-5p inhibitor treatment. Moreover, luciferase reporter assays indicated that transfection of miR-30c-5p led to a marked reduction of luciferase activity, but had no effect on Bcl-2 3'-UTR mutated fragment. Mechanically, miR-30c-5p promoted the activation of caspase 3 and caspase 9 in glioma cells. Furthermore, miR-30c-5p promoted apoptosis and inhibited colony formation and migration, and knockdown of Bcl2 further increased the number of apoptotic cells and suppressed colony formation and migration of glioma cells. By contrast, miR-30c-5p inhibitors decreased apoptosis and increased colony formation and migration, and restored Bcl2 expression further suppressed glioma cell apoptosis and enhanced colony formation and migration. Conclusions These results demonstrated that miR-30c-5p regulated growth, apoptosis and migration in glioma cells by targeting Bcl2, suggesting that miR-30c-5p might serve as a novel target for glioma therapy.

Non-small cell lung cancer (NSCLC) is associated with high mortality rates worldwide. The costimulatory molecule, B7-H4, a member of the B7 family, plays an important role in immune regulation, mainly by inhibiting the proliferation of T cells to achieve a negative regulatory T cell immune response. The mechanism of action of B7-H4 in non-small cell lung cancer is unknown at present. Tumor tissues from 71 patients subjected to radical pneumonectomy were examined, along with NSCLC cells and BALB/c mice. Among the 71 NSCLC cases, overall and recurrence-free survival rates were significantly lower in those displaying high B7-H4 expression. Mechanistic analyses showed that B7-H4 promoted the growth and metastasis of non-small cell lung cancer tumor tissues in mice through effects on CD8+ T cell apoptosis. Data from western blot experiments further suggested that B7-H4 induced CD8+ T cell death, both in vitro and in vivo, and affecting the PD-1/Stat3 pathway and promoting immune escape of tumor cells. Our collective findings support the potential utility of B7-H4 gene expression as a marker of NSCLC prognosis and provide a novel strategy for targeted therapy.

To study the reinforcement effect and the mechanical characteristic of micropiles in supporting soil landslide under step-loadings, two soil landslide models were built: the reinforced landslide with three rows of micropiles and the unreinforced landslide. The mechanical characteristics and failure mode of micropiles under step-loadings were analyzed by monitoring the displacement of landslides and pile-tops, and the strain of micropiles. The results indicate that the bearing capacity of the reinforced landslide with micropiles is increased by three times compared to the unreinforced landslide. Micropile plays a major role in anti-bending and shear-resistance. The failure pattern of micropiles shows reverse “S”-type. The destruction area of the back pile and the middle pile mainly distributes in the area that 1.7–3.1 times pile diameter above the sliding surface and the area that 1.7–2.9 times pile diameter below the sliding surface. The destruction area of the front pile mainly distributes in the area that 2.6–4.3 times pile diameter above the sliding surface and 1.7–2.9 times pile diameter below the sliding surface, and destructiveness of the front pile is less than that of the other two rows of piles. Before the landslide enters the unstable status, the bending moments of different rows of piles have not much difference. However, when the landslide is at the completely failure state, the back pile and the middle pile play more important role than the front pile.

The inhibition of alpha-ketoglutarate (α-KG)-dependent dioxygenases is thought to contribute to isocitrate dehydrogenase (IDH) mutation-derived malignancy. Herein, we aim to thoroughly investigate the expression pattern and prognostic significance of genes encoding α-KG-dependent enzymes for lower-grade glioma (LGG) patients. In this retrospective study, a total of 775 LGG patients were enrolled. The generalized linear model, least absolute shrinkage and selection operator Cox regression, and nomogram were applied to identify the enzyme-based signature. With the use of gene set enrichment analysis and Gene Ontology, the probable molecular abnormalities underlying high-risk patients were investigated. By comprehensively analyzing mRNA data, we observed that 41 genes were differentially expressed between IDHMUT and IDHWT LGG patients. A risk signature comprising 10 genes, which could divide samples into high- and low-risk groups of distinct prognoses, was developed and independently validated. This enzyme-based signature was indicative of a more malignant phenotype. The nomogram model incorporating the risk signature, molecular biomarkers, and clinicopathological parameters proved the incremental utility of the α-KG-dependent signature by achieving a more accurate prediction impact. Our study demonstrates that the α-KG-dependent enzyme-encoding genes were differentially expressed in relation to the IDH phenotype and may serve as a promising indicator for clinical outcomes of LGG patients.

Background: Brain metastases (BMs) are the most common central nervous system (CNS) malignant tumors, with rapid disease progression and extremely poor prognosis. The heterogeneity between primary tumors and BMs leads to the divergent efficacy of the adjuvant therapy response to primary tumors and BMs. However, the extent of heterogeneity between primary tumors and BMs, and the evolutionary process remains little known. Methods: To deeply insight the extent of inter-tumor heterogeneity at single-patient level and the process of these evolutions, we retrospectively analyzed a total of 26 tumor samples from 11 patients with matched primary tumors and BMs. One patient underwent four times brain metastatic lesion surgery with diverse locations and one operation for the primary lesion. The genomic and immune heterogeneity between primary tumors and BMs was evaluated by utilizing the whole-exome sequencing (WESeq) and immunohistochemical analysis.Results: In addition to inheriting genomic phenotype and molecular phenotype from the primary tumors, massive unique genomic phenotype and molecular phenotype were also observed in BMs, which revealed unimaginable complexity of tumor evolution and extensive heterogeneity among lesions at single-patient level. Our study also verified that the expression level of immune checkpoints-related molecule Programmed Death-Ligand 1 (PD-L1) (P = 0.0013) and the density of tumor-infiltrating lymphocytes (TILs) (P = 0.0248) in BMs were significantly lower than that in paired primary tumors. Additionally, tumor microvascular density (MVD) and tumor invasiveness were also differed between primary tumors and paired BMs, indicating that temporal and spatial diversity profoundly contributes to the evolution of BMs heterogeneity.Conclusion: We verified the significance of temporal and spatial factors to the evolution of tumor heterogeneity by multi-dimensional analysis of matched primary tumors and BMs, which also provided novel insight for formulating individualized treatment strategies of BMs.

Hepatitis C virus (HCV) is a major cause of chronic liver disease, which affects 2-3% of the world population. Until now, the early detection of HCV has been a great challenge, especially for those who live in developing countries. In this study, we developed a novel and ultrasensitive assay for the detection of HCV RNA based on the reduced graphene oxide nanosheets (rGONS) and hybridization chain reaction (HCR) amplification technique. This detection system contains a pair of single fluorophore-labeled hairpin probes that can freely exist in the solution in the absence of target RNA. The introduction of target RNA can robustly trigger a HCR with the two probes and produce long nanowires containing a double-stranded structure. The weak adsorption to rGONS makes the long nanowires emit a strong fluorescence. Using this enzyme-free amplification strategy, we developed a new method for the HCV RNA assay with a detection limit of 10 fM, which is far more sensitive than the common GO-based fluorescence method. Furthermore, the new method exhibits high selectivity for the discrimination of perfectly complementary and mismatched sequences. Finally, the new method was successfully used as a HCV RNA assay in biological samples with a strong anti-interference capability in complicated environments. In summary, these remarkable characteristics of the new method highlight its potential use in a clinical sample primary screening.

A series of shaking table tests were conducted on unreinforced and micropiles-reinforced soil landslides to investigate the seismic response of micropiles-reinforced landslide and the dynamic mechanical behaviour of micropiles in landslide under earthquake. The test results indicate that: (1) The micropiles can raise the anti-seismic performance of landslide, and it can efficiently delay the development of earthquake landslide hazards. (2) After earthquake, the flexural failure of micropile mainly occurs in the area that 1.4–4 times pile diameter above the sliding surface and the area that 1.4–3.4 times pile diameter below the sliding surface. (3) After reinforced by micropiles, the acceleration response on the slope surface of landslide can be decreased, especially the slope toe. (4) Both thrust of sliding mass and resistance of sliding bed are mainly concentrated near the sliding surface. The dynamic bending moment distributions of the three rows of micropiles present as reverse ‘S’. The maximum bending moment of the pile above sliding surface is located at the point that 3.7 times pile diameter above sliding surface, and the maximum bending moment of the pile below sliding surface is located to the point that 1.43 times pile diameter below sliding surface.

Compression anchor is a good anchorage structure in the slope and landslide project. Compared with tension anchor, it is a relatively new retaining structure, and the studies on compression anchor under seismic loadings are very limited. To investigate the seismic behavior of compression anchor under earthquake and get it to work much better in an earthquake-prone area, based on the shaking table test, a soil model slope reinforced with compression anchor and frame beam was carried out. In the test, the smooth steel bar and Q235 steel plate ring were used to simulate the anchor tendon and bearing plate of the compression anchor. The testing results indicate that the anchoring slope shows an obvious amplification effect on input seismic waves in both time domain and frequency domain, and the amplification effect of the acceleration response increases along the slope height. The axial force of the compression anchor is relatively uniform along the length of the anchor. As for different anchors in the same column, at 0.05~0.2 g, the axial forces of all anchors show tiny values, and the axial force of the bottom anchor is slightly greater than that of other anchors. The axial force of the bottom anchor is about 2.1~2.8 times that of the other anchors. At 0.3~0.6 g, the axial forces of the upper two anchors and the bottom anchor increase rapidly, while the axial forces of the other two middle anchors increase a little. The axial force ratio of the five rows of anchors from the bottom to the top at 0.3 g, 0.4 g, and 0.6 g is 4.2:1:1:2.1:2.9, 4.2:1:0.9:3.1:3.4, and 3.6:1:0.8:3.2:3.8, respectively. As for different beams, at 0.05~0.2 g, the bending moments of all beams show tiny values, and the bending moment of the bottom beam is slightly greater than that of other four beams. At 0.3~0.6 g, the bending moment of the top beam plays a more and more vital role, while the bending moment of the other four beams shows little variation.

It has been certified that long noncoding RNAs (lncRNAs) are crucial regulators in the progression of various human cancers. snoRNA host gene 6 (SNHG6) has been uncovered to affect the initial stage and tumorigenesis of hepatocellular carcinoma. Nevertheless, the expression pattern and biological role of SNHG6 in glioma still need to be investigated. The study aims to investigate the expression pattern, biological role and the potential mechanism of SNHG6 in glioma. In this study, the high expression of SNHG6 was tested in both glioma tissues and glioma cells. The correlation between expression levels of SNHG6 and the overall survival of glioma patients was demonstrated by using Kaplan Meier method analysis. Next, gain of function assays revealed that overexpression of SNHG6 can promote the formation of malignant phenotype of 1800 cell. However, results of loss-of-function assays revealed that silenced SNHG6 exerted the inhibitory function on glioma cell growth. Flow cytometric analysis was performed in glioma cells to detect the anti-oncogenic effects of silenced SNHG6 on cell cycle and apoptosis. Finally, we identified that p21 was involved in glioma cell proliferation after SNHG6 was downregulated. Taken together, we concluded that SNHG6 is a regulator and a potential therapeutic target in glioma.

A new fluorometric method is delineated for the detection of RNase H activity by combining DNAzyme with reduced graphene oxide (rGO). In the absence of RNase H, the fluorescence of FAM-labeled probe is quenched due to the strong adsorption on the rGO. The presence of RNase H can release the active DNAzyme from the DNA-RNA chimeric strand. This triggers the cleavage of the signal probe at the rA site with the help of the cofactor Mg2+. The recycle cleavage can directly result in the amplified signal emitted by the FAM-labeled short fragment. The method allows the activity of RNase H to be detected in a linear range of 0.01 to 5 U·mL−1. The detection limit of 0.018 U·mL−1 is calculated by the principle of three-time standard deviation over the blank signal. Then, RNase H-targeting natural compounds were screened for their inhibitory action. Among the investigated compounds, five were screened as RNase H inhibitors in a concentration-dependent manner, and 4 compounds were identified as activators. Finally, the method was reliably used for discriminating the difference of RNase H activity in human serum. It is found that RNase H activity was upregulated in patients with hepatitis C virus infection.

The neurogenin (Ngn) transcription factors control early neurogenesis and neurite outgrowth in mammalian cortex. In contrast to their proneural activity, their function in neurite growth is poorly understood. Drosophila has a single predicted Ngn homolog, Tap, of unknown function. Here we show that Tap is not a proneural protein in Drosophila but is required for proper axonal growth and guidance of neurons of the mushroom body, a neuropile required for associative learning and memory. Genetic and expression analyses suggest that Tap inhibits excessive axonal growth by fine regulation of the levels of the Wnt signaling adaptor protein Dishevelled., info:eu-repo/semantics/published

In this paper, the mechanical properties of Engineered Cementitious Composites (ECC) with two different fly ash contents after sub-elevated temperature exposure were investigated systematically at multiple scales. At composite level, uniaxial compressive and tensile tests were performed on specimens underwent 20 °C, 50 °C, 100 °C, and 200 °C exposure. The results indicate that, as temperature rise from 20 °C to 200 °C, the compressive strength of both ECC mixtures increased due to the refinement of the matrix pore structure. In term of tensile properties, after exposed to 100 °C and 200 °C, ECC M1 with relatively lower fly ash content shows a substantial reduction in strain capacity, but still retaining strain-hardening behavior, whereas ECC M2 with higher fly ash content exhibits a comparable or even enhanced tensile strength and strain capacity. At micro-scale, ECC matrix toughness was increased and PVA fiber strength was degraded under 200 °C exposure, which deteriorates the tensile properties of ECCs. However, the above negative effects are diminished due to the reduced fiber/matrix interfacial bond. Furthermore, the rather low matrix toughness and fiber/matrix interfacial bond associated with high fly ash content are the main reasons for the highly remained ductility in ECC M2 after sub-elevated temperature exposure.

Objective The aim of this research was to compare the functional outcome and safety between supraorbital keyhole approach (SKA) and pterional keyhole approach (PKA) for clipping intracranial aneurysms. Methods This is a retrospective study involving 318 patients with a total of 365 aneurysms who underwent keyhole surgery, comprising 195 cases in SKA group and 123 cases in PKA group. The outcome measures include Glasgow Outcome Scale, complete clipping rate, adverse events incidence, operation view angle, working distance, and surgical incision condition. Results Of a total of 356 aneurysms that were clipped and 9 trapped, no significant difference was observed in Glasgow Outcome Scale score, adverse events incidence, or complete clipping rate between the SKA and PKA groups. The distance from skin incision to anterior clinoid process was 5.87 ± 0.24 cm in SKA and 5.12 ± 0.27 cm in PKA. The operation view angle (from midline to the operating channel in sagittal plane) was 30°–40° in the SKA group and 60°–68° in the PKA group. Conclusions Our research demonstrates that both SKA and PKA are safe and effective for most anterior circulation aneurysms and parts of posterior circulation aneurysms. The SKA exposures aneurysm better on deep and sagittal directions and is more suitable for clipping aneurysms by the contralateral approach due to the short distance. The PKA has a good exposure on the neck of aneurysm with dorsal direction of parent artery and can be used to evacuate hematoma in the temporal lobe when clipping the aneurysm. Integrating multimodal 3-dimensional images could help neurosurgeon in selecting an appropriate and effective approach.

For the influence of corrosive environment, the reliability index of reinforced concrete structure will decrease in design reference period. A method of time-variant reliability is proposed based on method of moments. The resistance and load effect are described as random process. The random process is transformed into random variable by the method of equivalent resistance method. The time-variant reliability is carried on in design reference period accurately with consideration of characteristic of random variables which can provide supports for the management departments to take appropriate measures to reduce economic loss in the engineering circles.

The Neurogenin (Ngn) transcription factors control early neurogenesis and neurite outgrowth in mammalian cortex. In contrast to their proneural activity, their function in neurite growth is poorly understood.Drosophilahas a single predicted Ngn homologue called Tap, whose function is completely unknown. Here we show that Tap is not a proneural protein inDrosophilabut is required for proper axonal growth and guidance of neurons of the mushroom body (MB), a neuropile required for associative learning and memory. Genetic and expression analyses suggest that Tap inhibits excessive axonal growth by fine regulation of the levels of the Wnt signaling adaptor protein, Dishevelled.SummaryTheDrosophilaNeurogenin homologue, Target of Pox neuro (Tap), prevents axonal overgrowth by regulating the Wnt Planar Cell Polarity pathway adaptor protein Dishevelled.

Neurogenesis is initiated by the transient expression of the highly conserved proneural proteins, bHLH transcriptional regulators. Here, we discover a conserved post-translational switch governing the duration of proneural protein activity that is required for proper neuronal development. Phosphorylation of a single Serine at the same position in Scute and Atonal proneural proteins governs the transition from active to inactive forms by regulating DNA binding. The equivalent Neurogenin2 Threonine also regulates DNA binding and proneural activity in the developing mammalian neocortex. Using genome editing in Drosophila, we show that Atonal outlives its mRNA but is inactivated by phosphorylation. Inhibiting the phosphorylation of the conserved proneural Serine causes quantitative changes in expression dynamics and target gene expression resulting in neuronal number and fate defects. Strikingly, even a subtle change from Serine to Threonine appears to shift the duration of Atonal activity in vivo, resulting in neuronal fate defects. publisher: Elsevier articletitle: Post-translational Control of the Temporal Dynamics of Transcription Factor Activity Regulates Neurogenesis journaltitle: Cell articlelink: http://dx.doi.org/10.1016/j.cell.2015.12.048 content_type: article copyright: Copyright © 2016 Elsevier Inc. All rights reserved. ispartof: Cell vol:164 issue:3 pages:460-475 ispartof: location:United States status: published

The production of neurons, astrocytes and oligodendrocytes is regulated by a group of transcription factors, which determine cell fates and specify subtype identities in the nervous system. Here we focus on profiling the distinct roles of Neurogenin (Ngn or Neurog) family members during the neuronal development. Ngn proteins are tightly regulated to be expressed at defined times and positions of different progenitor cell pools. In addition to their well-elucidated proneural function, Ngn proteins play various critical roles to specify or maintain cell fate and regulate neurite outgrowth and targeting in the central nervous system. Finally, Ngns have been associated with neuronal disorders. Therefore understanding the function and regulation of Ngns will not only improve the understanding of the molecular mechanism underlying the development of nervous system, but may also provide insight into neuronal disease.

1. Bronchoconstriction can be evoked by electrical stimulation of the vagus nerves in the presence of atropine. We have used novel, highly selective tachykinin receptor antagonists, together with a procedure for on-line, breath-by-breath analysis of total lung resistance (R(L), subtractor method) and dynamic lung compliance (C(dyn)), to investigate the role of tachykinins in this response in anaesthetized, paralysed guinea-pigs. 2. In the presence of 1 mg/kg phosphoramidon (a neutral endopeptidase inhibitor), CP 96345 (the non-peptide NK1 selective antagonist) at 200 nmol/kg had no effect on the increase in R(L) caused by vagal stimulation, but significantly inhibited the associated decrease in C(dyn). 3. The NK2 selective antagonist, MDL 29913 (1 mu mol/kg), significantly antagonized the changes in both R(L) and C(dyn). In the absence of phosphoramidon, MDL 29913 again significantly inhibited the changes in R(L) and C(dyn) although CP 96345 no longer had any effect. 4. The non-peptide NK2 selective antagonist, SR 48968 (100 nmol/kg), also effectively inhibited the responses to vagal stimulation and was more potent than MDL 29913. 5. These results emphasize the importance of the NK2 receptor system in mediating non-cholinergic bronchoconstriction evoked by vagal stimulation in the guinea-pig.

The present study aimed to compare the reflex bronchoconstrictor effects of upper airway applications of dry airstreams with equivalent cooling produced by cold isotonic and hypertonic saline solutions in anaesthetized rabbits. A method of on-line, breath by breath analysis of lung mechanics was used. The changes in total lung resistance (RL) and dynamic lung compliance (Cdyn) produced by dry airstreams and by cold hypertonic saline were greater than those induced by cold isotonic saline. Warm isotonic saline and supersaturated airstreams were ineffective. These results implicate osmotic disturbances in the response to dry airstreams. The response to dry airflow was abolished by section of the superior laryngeal nerves, by vagotomy and by atropine, suggesting a central cholinergic reflex mechanism is involved. Phosphoramidon had no effect, indicating that tachykinins are unlikely to play a role in this particular reflex.

Wilms tumor (WT) is a genetically heterogeneous childhood kidney tumor. Several genetic mutations have been identified in WT patients, including inactivation of WTX, somatic stabilizing CTNNB1, and p53 mutations. However, the molecular mechanisms in tumorigenesis remain largely unexplored. Stat3 is a transcription factor that can promote oncogenesis. Stat3 activation is commonly viewed as crucial for multiple tumor proliferation and metastasis. We show that Stat3 is highly activated in Wilms tumor tissues compared to those in adjacent tissues. IL-6 treatment or transfection of a constitutively activated Stat3 in G401 cells promotes cell proliferation. At the molecular level, we further elucidate that Stat3 inhibits WTX expression through up-regulation of microRNA-370. Our results suggest that Stat3/miR-370/WTX regulatory axis might be a critical mechanism in Wilms tumor cells.

Summary: Tachykinins are potent bronchoconstrictors. Here we have used novel highly selective agonists and antagonists to characterize tachykinin receptors mediating bronchoconstriction and hypotension in anaesthetized, paralysed guinea-pigs. Total lung resistance ( R L ) and dynamic lung compliance ( C dyn ) were measured using an online, breath-by-breath method to analyse lung mechanics. The effects of iv injections of neuropeptide γ (NPγ), [Sar 9 ,Met(O 2 ) 11 ]-SP (Sar-SP) (NK-1 agonist), [Lys 5 ,MeLeu 9 ,Nle 10 ]-NKA(4-10) (Lys-NKA) (NK-2 agonist) and senktide (NK-3 agonist) were compared, before and after iv injection of the NK-1 antagonist CP 96345 or the NK-2 antagonists MDL 29913 and SR 48968. NPγ, Sar-SP and Lys-NKA, but not senktide, increased R L and decreased C dyn , accompanied by a fall in blood pressure (BP). The potency for increase of R L was Lys-NKA>NPγ≃Sar-SP, for reduction of C dyn was Lys-NKA=Sar-SP>NPγ, and for fall in BP was Sar-SP>NPγ>>Lys-NKA. CP 96345 (50 nmol/kg & 200 nmol/kg) significantly antagonized the changes in R L , C dyn and BP induced by Sar-SP, and the fall in BP (but not changes in lung mechanics) in response to NPγ, but did not alter any responses to Lys-NKA. In contrast, MDL 29913 (200 nmol/kg & 10 μmol/kg) weakly and SR 48968 (100 nmol/kg) potently inhibited the effects on R L and C dyn induced by Lys-NKA and NPγ. Our results show that both NK-1 and NK-2 receptors are represented in guinea-pig airways. MDL 29913 is a weak NK-2 antagonist in guinea-pig airways compared to SR 48968. In addition, NK-1 but not NK-2 receptors appear to be important in mediating the hypotensive response to these tachykinins. In spite of its ability to cause a fall in BP via NK-1 receptors, NPγ exerted its effects on R L and C dyn solely via NK-2 receptors.

The effects of iv injections of two endogenous tachykinins, substance P (SP) and neuropeptide γ and the highly selective tachykinin agonists [Sar 9 ,Met(O 2 ) 11 ]-SP, [Lys 5 ,MeLeu 9 ,Nle 10 ]-NKA(4–10) and senktide, on total lung resistance (R L ), dynamic lung compliance (C dyn ) and systemic blood pressure, were compared in the anaesthetized rabbit. Senktide, the NK-3 receptor selective agonist, had no effect on R L , C dyn or blood pressure. The other four agonists caused dose-dependent increases in R L and C dyn , with [Sar 9 ,Met(O 2 ) 11 ]-SP being the most potent agonist in producing changes in the absence of phosphoramidon. This suggested that NK-1 receptors play an important role in these responses. [Sar 9 ,Met(O 2 ) 11 ]-SP, SP and neuropeptide γ also decreased blood pressure. Phosphoramidon (1 mg/kg) potentiated the changes in R L and C dyn evoked by [Sar 9 ,Met(O 2 ) 11 ]-SP and SP, with very marked enhancement of responses to neuropeptide γ. Responses to [Lys 5 ,MeLeu 9 ,Nle 10 ]-NKA(4–10) were unaffected, suggesting that this NK-2 selective agonist may not be catabolized by neutral endopeptidase (NEP). In the presence of phosphoramidon, the non-peptide tachykinin NK-1 receptor selective antagonist CP 96345 (80 nmol/kg) reduced all responses to [Sar 9 ,Met(O 2 ) 11 ]-SP and SP, whereas the NK-2 selective antagonist SR 48968 (40 nmol/kg) inhibited the bronchomotor but not the vasodepressor responses to neuropeptide γ and [Lys 5 ,MeLeu 9 ,Nle 10 ]-NKA(4–10). The fall in blood pressure induced by neuropeptide γ was diminished by CP 96345, whereas bronchoconstriction was unaffected, indicating possible differences in NK-1 receptors in the vasculature and airways. Electrical stimulation of the distal ends of vagus nerves caused increases in R L which were abolished by atropine (1 mg/kg).

We investigated the corticocortical connections between area 21a and ipsilateral areas 17 and 18 in the cat. The anterograde/retrograde fluorescent tracer tetramethyl-rhodamine conjugated to dextran (Fluoro-Ruby) was injected into area 21a of the anaesthetized cat. Cell bodies labelled retrogradely from area 21a were consistently observed in both areas 17 and 18, primarily located in the supragranular layers of cortex where they formed discrete patches of cells. Similar numbers of cell bodies were labelled retrogradely in areas 17 and 18 of each animal. Our data are also consistent with previous reports of a reciprocal projection from area 21a back to areas 17 and 18 terminating principally in infragranular cortical layers.