Characterization of a CNS penetrant, selective M1 muscarinic receptor agonist, 77-LH-28-1.

BACKGROUND AND PURPOSE: M1 muscarinic ACh receptors (mAChRs) represent an attractive drug target for the treatment of cognitive deficits associated with diseases such as Alzheimer's disease and schizophrenia. However, the discovery of subtype-selective mAChR agonists has been hampered by the high degree of conservation of the orthosteric ACh-binding site among mAChR subtypes. The advent of functional screening assays has enabled the identification of agonists such as AC-42 (4-n-butyl-1-[4-(2-methylphenyl)-4-oxo-1-butyl]-piperidine), which bind to an allosteric site and selectively activate the M(1) mAChR subtype. However, studies with this compound have been limited to recombinantly expressed mAChRs. EXPERIMENTAL APPROACH: In this study, we have compared the pharmacological profile of AC-42 and a close structural analogue, 77-LH-28-1 (1-[3-(4-butyl-1-piperidinyl)propyl]-3,4-dihydro-2(1H)-quinolinone) at human recombinant, and rat native, mAChRs by calcium mobilization, inositol phosphate accumulation and both in vitro and in vivo electrophysiology. KEY RESULTS: Calcium mobilization and inositol phosphate accumulation assays revealed that both AC-42 and 77-LH-28-1 display high selectivity to activate the M1 mAChR over other mAChR subtypes. Furthermore, 77-LH-28-1, but not AC-42, acted as an agonist at rat hippocampal M1 receptors, as demonstrated by its ability to increase cell firing and initiate gamma frequency network oscillations. Finally, 77-LH-28-1 stimulated cell firing in the rat hippocampus in vivo following subcutaneous administration. CONCLUSIONS AND IMPLICATIONS: These data suggest that 77-LH-28-1 is a potent, selective, bioavailable and brain-penetrant agonist at the M1 mAChR and therefore that it represents a better tool than AC-42, with which to study the pharmacology of the M1 mAChR.

Segmental nature of the porcine liver and its potential as a model for experimental partial hepatectomy.

BACKGROUND: In-depth knowledge of pig liver anatomy allows potential research into segmental liver resections and hepatic regeneration, as well as liver transplantation techniques. The segmental anatomy, however, remains largely unknown. This study aimed to delineate the segmental anatomy of the porcine liver in comparison with that of the human. METHODS: The segmental anatomy of the porcine liver was determined using acrylic injection casting of ex vivo pig livers, allowing the arterial, venous and biliary supply to be visualized directly. This was correlated using multi-slice computed tomography (CT) and three-dimensional reconstructions. RESULTS: Although the external morphology of the porcine liver differs from that of the human, the segmental anatomy is remarkably similar in term of its vascularity and biliary tree. CONCLUSION: Acrylic casting of the porcine liver accurately delineates the vascular and biliary anatomy, and is a useful tool for performing experimental liver surgery. The similarities between porcine and human segmental anatomy allow domestic swine to be used as a comparable model. Three-dimensional CT reconstructions can also accurately visualize the anatomy and may be used to perform virtual surgery, or to assess segmental volumes.

Pharmacological characterization of Ro 63-1908 (1-[2-(4-hydroxy-phenoxy)-ethyl]-4-(4-methyl-benzyl)-piperidin-4-ol), a novel subtype-selective N-methyl-D-aspartate antagonist.

Ro 63-1908, 1-[2-(4-hydroxy-phenoxy)-ethyl]-4-(4-methyl-benzyl)-piperidin-4-ol, is a novel subtype-selective N-methyl-D-aspartate (NMDA) antagonist that has been characterized in vitro and in vivo. Ro 63-1908 inhibited [(3)H]dizocilpine ((3)H-MK-801) binding in a biphasic manner with IC(50) values of 0.002 and 97 microM for the high- and low-affinity sites, respectively. Ro 63-1908 selectively blocked recombinant receptors expressed in Xenopus oocytes containing NR1C + NR2B subunits with an IC(50) of 0.003 microM and those containing NR1C + NR2A subunits with an IC(50) of >100 microM, thus demonstrating greater than 20,000-fold selectivity for the recombinant receptors expressing NR1C + NR2B. Ro 63-1908 blocked these NMDA NR2B-subtype receptors in an activity-dependent manner. Ro 63-1908 was neuroprotective against glutamate-induced toxicity and against oxygen/glucose deprivation-induced toxicity in vitro with IC(50) values of 0.68 and 0.06 microM, respectively. Thus, the in vitro pharmacological characterization demonstrated that Ro 63-1908 was a potent and highly selective antagonist of the NR2B subtype of NMDA receptors. Ro 63-1908 was active against sound-induced seizures (ED(50) = 4.5 mg/kg i.p. when administered 30 min beforehand) in DBA/2 mice. The dose required to give a full anticonvulsant effect did not produce a deficit in the Rotarod test. NMDA-induced seizures were also inhibited by Ro 63-1908 with an ED(50) of 2.31 mg/kg i.v. when administered 15 min before testing. Ro 63-1908 gave a dose-related neuroprotective effect against cortical damage in a model of permanent focal ischemia. Maximum protection of 39% was seen at a plasma concentration of 450 ng/ml. There were, however, no adverse cardiovascular or CNS side-effects seen at this dosing level.

Use of tonsil size in the evaluation of obstructive sleep apnoea.

AIMS: (1) To determine the extent to which tonsil size contributes to the severity of obstructive sleep apnoea (OSA) in children; and (2) to assess the use of tonsillar-pharyngeal (TP) ratio in differentiating patients with different severity of OSA. METHODS: Lateral neck radiograph was performed on 35 children referred consecutively to a university paediatric chest clinic for suspected OSA secondary to tonsillar hypertrophy. The tonsil size was determined by measuring the TP ratio on the radiographs. The severity of OSA was assessed by overnight polysomnography. RESULTS: A total of 24 boys and 11 girls (median age 6.2 years) were studied. All presented with symptoms of OSA, and tonsillar hypertrophy was detected on clinical examination. The median apnoea-hypopnoea index (AHI) was 16.93 (interquartile range: 8.41 to 28.29). The median TP ratio was 0.76 (interquartile range: 0.65 to 0.80). AHI was positively correlated with the TP ratio. The clinical tonsil size did not correlate with the AHI or the TP ratio. Using a TP ratio of 0.479, the sensitivity and specificity in predicting cases with moderate/severe OSA (AHI >10) were 95.8% and 81.8% respectively, while the positive and negative predictive values were 92.0% and 90.0% respectively. CONCLUSIONS: Results show that in a population of children with OSA, tonsillar hypertrophy as assessed by lateral neck radiograph correlates positively with the severity of obstructive sleep apnoea. The TP ratio has high sensitivity and specificity in predicting those with moderate/severe disease and this feature may be used as a clinical screening method in prioritising patients with OSA for further assessment.

A combined pharmacological and genetic approach to investigate the role of orphanin FQ in learning and memory.

Using a combination of the selective opioid receptor-like1 (ORL1) receptor agonist, Ro 64-6198, and orphanin FQ/nociceptin (OFQ/N) peptide knockout (KO) mice, the influence of OFQ/N on cognition has been studied in the rodent. In wild type, C57BL/6J mice, Ro 64-6198 (0.3-1 mg/kg i.p.) impaired the acquisition of spatial learning in the Morris water maze, although a mild neurological impairment was evident which complicated precise interpretation. In Lister hooded rats, Ro 64-6198 (6 mg/kg i.p.) produced delay dependent impairments in rats performing either a delayed matching or a delayed nonmatching to position task with only a modest (< 20%) effect on omissions - an effect consistent with a short-term memory impairment. Electrophysiological studies demonstrated an inhibitory effect of OFQ/N on LTP recorded from the CA1 region of wild type mice, but not in ORL1 receptor knockout mice. In contrast to the ORL1 agonist, mice deficient in the OFQ/N peptide showed some evidence of improved spatial learning, fear conditioning and passive avoidance retention. However, CA1 LTP was similar between OFQ/N peptide KO mice and wild type controls. Subsequent receptor radioautography studies demonstrated the presence of ORL1 receptors within various regions of the medial temporal lobe system: i.e. CA1, dentate gyrus molecular layer, subiculum, perirhinal cortex. Taken together, these results suggest a bi-directional effect of OFQ/N containing systems on aspects of cognitive behaviour, particularly those elements associated with hippocampal function. This is consistent with a likely modulatory role of OFQ/N on hippocampal and associated cortical circuitry.

Positive allosteric modulators of metabotropic glutamate 1 receptor: characterization, mechanism of action, and binding site.

We have identified two chemical series of compounds acting as selective positive allosteric modulators (enhancers) of native and recombinant metabotropic glutamate 1 (mGlu1) receptors. These compounds did not directly activate mGlu1 receptors but markedly potentiated agonist-stimulated responses, increasing potency and maximum efficacy. Binding of these compounds increased the affinity of a radiolabeled glutamate-site agonist at its extracellular N-terminal binding site. Chimeric and mutated receptors were used to localize amino acids in the receptor transmembrane region critical for these enhancing properties. Finally, the compounds potentiated synaptically evoked mGlu1 receptor responses in rat brain slices. The discovery of selective positive allosteric modulators of mGlu1 receptors opens up the possibility to develop a similar class of compounds for other family 3 G protein-coupled receptors.

Lateralized carotid artery: an unusual cause of pulsatile tinnitus.

We describe the case of a patient who had a pulsatile tinnitus that was caused by a laterally displaced internal carotid artery. Her condition was treated with the use of a hearing did, which suppressed the tinnitus. We also review the literature on laterally displaced internal carotid arteries, and discuss their differentiation from a congenitally aberrant artery.

X-linked sideroblastic anaemia with ataxia: another mitochondrial disease?

OBJECTIVES: The syndrome of X-linked sideroblastic anaemia with ataxia is rare, described only twice in the literature. The aim was to obtain clinical neurological and haematological data about this rare syndrome throughout adult life. METHODS: A family is described with two affected brothers and two affected maternal uncles. The family was evaluated clinically. Haematological investigations included full blood count, blood film, iron studies, free erythrocyte protoporphyrin (FEP) concentrations and a bone marrow examination where possible. RESULTS: Core neurological features included motor delay, ataxia evident from early childhood, and dysarthria. Neurological features were non-progressive until the fifth decade when slow progression became evident. Some family members showed mild spasticity. Patients usually have a mild asymptomatic anaemia or a borderline decreased mean corpuscular volume. Blood film examination showed Pappenheimer bodies. Bone marrow examination showed ring sideroblasts, indicating raised erythrocyte iron. Free erythrocyte protoporphyrin (FEP) concentrations were raised. CONCLUSIONS: Haematological features are subtle and can be easily overlooked, and individual patients may not display all the abnormal features. X-linked ataxias are rare and incorrect genetic advice may be given if the diagnostic haematological features of X-linked sideroblastic anaemia are overlooked. Males with early onset ataxia should have a haematological evaluation including a blood film, with a bone marrow examination if abnormal blood count indices and measurement of FEP concentrations raise suspicion. The condition has parallels with Pearson's syndrome and Friedreich's ataxia. All three conditions are associated with mitochondrial iron handling defects and ataxia. The human ATP binding cassette gene (hABC7) is a candidate gene and requires further investigation.

Activity-dependent presynaptic autoinhibition by group II metabotropic glutamate receptors at the perforant path inputs to the dentate gyrus and CA1.

Pharmacological activation of metabotropic glutamate receptors (mGluRs) can inhibit synaptic transmission; however, relatively little evidence exists regarding the physiological conditions under which such autoreceptors are activated by synaptically released glutamate. Bath application of selective group II mGluR agonists profoundly inhibited field excitatory postsynaptic potentials (fEPSPs) evoked by stimulation of the perforant path inputs to both the mid-molecular layer of the dentate gyrus and the stratum lacunosum moleculare of the CA1. Application of the group II selective mGluR antagonist LY341495 resulted in an increase in the relative amplitude of a test fEPSP evoked 200 ms after a conditioning burst, but not after a single conditioning stimulus, in both pathways. Antagonist application also resulted in a marked increase in the relative amplitude of test population spikes evoked in the dentate gyrus following a conditioning burst. These observations are consistent with a presynaptic autoinhibitory action of group II metabotropic receptors that is revealed following burst stimulation of the pathway, consistent with their localisation in the preterminal zone. Activation of group II mGluRs during theta-gamma pattern discharge of projection neurones in the entorhinal cortex is likely to play an important role in the regulation of synaptic transmission and plasticity in the perforant pathway.

Community-acquired pneumonia--implementation of a prediction rule to guide selection of patients for outpatient treatment.

The aim of this study was to determine the outcomes of outpatient treatment of community-acquired pneumonia (CAP) when a prediction rule was followed by emergency physicians to guide the selection of patients. This was a prospective observational study conducted at the emergency department of a university-affiliated hospital in Hong Kong, China. A clinical prediction rule was implemented to guide the selection of patients with CAP for outpatient treatment. All subsequent hospitalizations gial presentation were recorded, and the reasons were assessed. The utilization of the observation unit was incorporated into the treatment algorithm. Of 72 patients with CAP followed up as outpatients, 60 (83.3%) were treated successfully, nine (12.5%) required subsequent hospitalization within 30 days, and three (4.2%) were lost to follow-up. None of the patients died, and none required admission to the intensive care unit. Factors associated with subsequent hospitalization include: tuberculosis, underlying malignancy, persistent fever, comorbidity (rheumatoid arthritis and severe osteoporosis), intravenous drug addiction and alcoholism. The observation ward was utilized in 10 (16.7%) patients successfully treated as outpatients. It is concluded that the prediction rule can be safely implemented as a guide for emergency physicians. The short-stay observation unit may be usefully employed for treating low-risk CAP.

Middle ear effusions after radiotherapy: correlation with pre-radiotherapy nasopharyngeal tumor patterns.

OBJECTIVE: The purpose of this study was to assess whether pretreatment tumor patterns of nasopharyngeal carcinoma (NPC) can predict the status of the middle ear after radiation treatment. MATERIALS AND METHODS: Pretreatment and follow-up magnetic resonance imaging (MRI) was performed in 32 patients (64 ears) who had radiation therapy for NPC. For the purpose of analysis, the ears were placed into their pre-radiation therapy tumor pattern groups and the presence of middle ear effusion (MEE) with regard to eustachian tube (ET) invasion or displacement was identified. RESULTS: MEEs were present in 31 (48.4%) ears after radiation therapy. All of the MEEs that resolved were in the preradiation therapy groups where tumor invasion of the eustachian tube was present irrespective of the amount of ET displacement. There was, however, no significant difference for resolved MEEs between ears with ET invasion or displacement (p = 0.32 and p = 0.71, respectively, Fisher's exact test). The MEEs occurred with significantly greater frequency in ears with minor ET displacement than in those with major ET displacement (p = 0.013, Fisher's exact test) as well as in previously normal ears compared with other groups (p = 0.008, Mann-Whitney U test). CONCLUSION: A pre-radiation therapy NPC tumor pattern was not found that clearly predicted the outcome of MEE after radiation treatment. The findings, however, suggest that approximately one third of MEEs in patients with invasion of ET or paratubal structures, irrespective of the amount of ET displacement, resolve after therapy. The MEEs that were present in ears with ET displacement and no invasion did not resolve despite reversal of the displacement after treatment, which suggests that ET position plays a less important role than invasion in the resolution of MEE. It is, however, difficult to be certain, because ears with invasion were invariably associated with ET displacement, and the contribution of radiation therapy to the development of MEE further complicates the issue.

Radiation induced sarcomas of the head and neck following radiotherapy for nasopharyngeal carcinoma.

AIM: To report the radiological findings of radiation induced sarcomas (RIS) in the head and neck following radiotherapy for nasopharyngeal carcinoma. MATERIALS AND METHODS: The clinical notes and radiological studies (MR n = 3, CT n = 4) of four patients were reviewed retrospectively. RESULTS: RIS developed 5 to 10 years following radiotherapy. Two patients had tumours arising from the alveolar process of the maxilla, one from the nasal cavity, and one patient had a tumour at two sites, involving the external auditory canal and the uvula. Three of the four patients had large tumours at diagnosis with a 3.5-6 cm predominately homogeneous soft tissue mass, complete destruction of bone and extensive local invasion. One was small and localized to the nasal turbinate. Radiation osteitis was identified in two of the four (50%) patients. CONCLUSION: The site of RIS following radiotherapy for NPC is variable but is invariably within the high dose zone of the radiotherapy. These sarcomas tend to present late with a large soft tissue mass. Radiation osteitis is not a constant feature. As surgery provides the only chance of cure, imaging has an important role in the pre-operative mapping of the extent of tumour. et al.

Intranasal anatomy of the nasolacrimal sac in endoscopic dacryocystorhinostomy.

Intranasal surface anatomy is fundamental to the technique of endoscopic dacryocystorhinostomy. In the current literature the lacrimal sac is described as being situated anterior to the anterior end of the middle turbinate with between 0% and 20% of the sac above the insertion of the middle turbinate on the lateral nasal wall (the axilla of the middle turbinate). The aim of this study was to use CT dacryocystograms (DCGs) and CT scans to establish the relationship of the lacrimal sac to the lateral nasal wall. Forty-seven individual lacrimal sacs were measured in relation to the common canaliculus, and 76 were measured in relation to the insertion of the middle turbinate. Measurements taken from the long axis of the sac showed the mean height of the sac above the middle turbinate insertion was 8.8 mm (SD = 0.2, 95% CI = 1.3) and below it was 4.1 mm (SD = 2.3, 95% CI = 1.1). The average measurement of the sac above the com-mon canaliculus on CT DCGs was 5.3 mm (SD = 1.7, 95% CI = 0.56), whereas the average measurement below the common canaliculus was 7.7 mm (SD = 2, 95% CI = 1.3) (n = 47 CT DCGs). The findings in this study show that a major portion of the sac is locat-ed above the insertion of the anterior end of the middle turbinate and, in addition, that a significant part of the sac lies above the entry point of the common canaliculus. Knowledge of these findings can ensure that the sac is adequately exposed during dacryocystorhinostomy by removal of sufficient bone and mucosa above the anterior insertion of the middle turbinate.

Differentiation of lymphadenopathy in different forms of carcinoma with Doppler sonography.

AIM: The aim of the study was to show whether it may be possible to characterize malignant nodes of different aetiologies with Doppler imaging. Non-Hodgkin's lymphoma (NHL) and nasopharyneal carcinoma (NPC) were used as the models. MATERIALS AND METHODS: A prospective study was undertaken using colour Doppler sonography in 60 enlarged peripheral lymph nodes of 34 patients with either lymphoma or NPC. The vascular distribution, peak systolic velocity (PSV), end diastolic velocity (EDV), resistivity index (RI) and the pulsatility index (PI) of the largest node on each side of the patient were investigated. RESULTS: The vascular distribution was predominantly 'capsular and central' or 'capsular' in 57 nodes, consistent with previous descriptions, but there was no significant difference in the mean PSV (lymphoma: 20.0 cm/s; NPC: 21.86 cm/s;P > 0.05) and mean EDV (lymphoma: 5.95 cm/s; NPC: 5.07 cm/s;P > 0.05). However, significant differences were shown in the mean RI (lymphoma: 0.71; NPC: 0.81;P < 0.05) and mean PI (lymphoma: 1.43; NPC: 1.88;P < 0.05) of these two groups of lymph nodes. CONCLUSIONS: There is significant difference in the Doppler waveform between malignant nodes affected by lymphoma or NPC, suggesting that further differentiation of malignant nodes may be possible.