Maintaining wakefulness is associated with a progressive increase in the need for sleep. This phenomenon has been linked to changes in synaptic function. The synaptic adhesion molecule Neuroligin-1 (NLG1) controls the activity and synaptic localization of N-methyl-Daspartate receptors, which activity is impaired by prolonged wakefulness. We here highlight that this pathway may underlie both the adverse effects of sleep loss on cognition and the subsequent changes in cortical synchrony. We found that the expression of specific Nlg1 transcript variants is changed by sleep deprivation in three mouse strains. These observations were associated with strainspecific changes in synaptic NLG1 protein content. Importantly, we showed that Nlg1 knockout mice are not able to sustain wakefulness and spend more time in nonrapid eye movement sleep than wild-type mice. These changes occurred with modifications in waking quality as exemplified by low theta/alpha activity during wakefulness and poor preference for social novelty, as well as altered delta synchrony during sleep. Finally, we identified a transcriptional pathway that could underlie the sleep/wake-dependent changes in Nlg1 expression and that involves clock transcription factors. We thus suggest that NLG1 is an element that contributes to the coupling of neuronal activity to sleep/wake regulation.ChIP | EEG | gene expression | sleep homeostasis | synaptic plasticity S leep is crucial for learning, memory, and other functions essential for proper functioning of the brain and body (1, 2). These functions have been associated with the sleep recovery process, which defines a level of pressure for sleep that increases with wakefulness and dissipates during sleep and that is reflected by changes in sleep intensity (3, 4). Sleep intensity is indexed by electroencephalographic (EEG) markers of neuronal synchrony in delta frequencies (1-4 Hz) measured during nonrapid eye movement (NREM) sleep (5). During wakefulness, mechanisms favoring desynchrony in the delta range predominate, and the brain can maintain cognition, whereas during sleep, events promoting network synchrony mostly take place with high delta activity thought to be permissive of recovery (3, 6). The sleep recovery process has been hypothesized to originate and contribute to the maintenance of both synaptic and network equilibrium (6-8). This notion is supported by the observation that specific plasticityrelated genes may be directly involved in regulating sleep need (9). Certain clock genes may also directly contribute, in a circadian-independent manner, to the sleep recovery process (5, 9). However, the mechanisms underlying the capacity and requirement of the brain to switch from an alert desynchronized state to an unconscious synchronized state remain elusive.Glutamate, the main excitatory neurotransmitter of the brain, can induce long-term modifications of synaptic transmission and, thus, changes in network connectivity. This is achieved mainly via glutamate's action on two types of receptors: N-methyl-D-aspa...

The bronchus sign on CT represents the presence of a bronchus leading directly to a peripheral pulmonary lesion. We investigated the value of this sign in predicting the results of transbronchial biopsy and brushing in 33 consecutive cases of proved peripheral bronchogenic carcinoma studied with thin-slice CT (2-mm-thick sections). The bronchus sign was seen on CT in 22 patients and was absent in 11. Transbronchial biopsy and brushing showed peripheral carcinoma in 13 (59%) of 22 patients in whom the bronchus sign was seen on CT and in only two (18%) of 11 patients in whom it was not seen. The difference is statistically significant (Fisher's exact test, p = .029). When analyzed by the order of involved bronchus, a 90% success rate of transbronchial biopsy and brushing was found in patients in whom the bronchus sign was seen at a fourthorder bronchus (p = .01). This compared with a success of 33% when the bronchus sign was seen at fifth-, sixth-, or seventh-order branches. Our results suggest that the bronchus sign at a fourth-order bronchus is valuable in predicting the success of transbronchial biopsy and brushing. The presence of the sign on CT may be useful in determining if the workup should include transbronchial biopsy and brushing or transthoracic needle aspiration in patients with peripheral lung lesions.

We have previously demonstrated that clock genes contribute to the homeostatic aspect of sleep regulation. Indeed, mutations in some clock genes modify the markers of sleep homeostasis and an increase in homeostatic sleep drive alters clock gene expression in the forebrain. Here, we investigate a possible mechanism by which sleep deprivation (SD) could alter clock gene expression by quantifying DNA-binding of the core-clock transcription factors CLOCK, NPAS2, and BMAL1 to the cis-regulatory sequences of target clock genes in mice. Using chromatin immunoprecipitation (ChIP), we first showed that, as reported for the liver, DNA-binding of CLOCK and BMAL1 to target clock genes changes in function of time-of-day in the cerebral cortex. Tissue extracts were collected at ZT0 (light onset), −6, −12, and −18, and DNA enrichment of E-box or E'-box containing sequences was measured by qPCR. CLOCK and BMAL1 binding to Cry1, Dbp, Per1, and Per2 depended on time-of-day, with maximum values reached at around ZT6. We then observed that SD, performed between ZT0 and −6, significantly decreased DNA-binding of CLOCK and BMAL1 to Dbp, consistent with the observed decrease in Dbp mRNA levels after SD. The DNA-binding of NPAS2 and BMAL1 to Per2 was also decreased by SD, although SD is known to increase Per2 expression in the cortex. DNA-binding to Per1 and Cry1 was not affected by SD. Our results show that the sleep-wake history can affect the clock molecular machinery directly at the level of chromatin binding thereby altering the cortical expression of Dbp and Per2 and likely other targets. Although the precise dynamics of the relationship between DNA-binding and mRNA expression, especially for Per2, remains elusive, the results also suggest that part of the reported circadian changes in DNA-binding of core clock components in tissues peripheral to the suprachiasmatic nuclei could, in fact, be sleep-wake driven.

Our results add further proof to the notion that clock genes are involved in sleep homeostasis. Because accumulating evidence directly links REV-ERBα to dopamine signaling the altered homeostatic regulation of sleep reported here are discussed in that context.

To investigate the value of computed tomography (CT) for depicting the relationship between carcinomatous solitary pulmonary nodules and the bronchial tree and predicting the results of various bronchoscopic biopsy techniques, the authors retrospectively reviewed CT scans from 27 consecutive patients with solitary pulmonary nodules associated with a positive bronchus sign. All patients underwent bronchoscopy and transbronchial biopsy. Macroscopic demonstration of the tumor-bronchi relationship was obtained in 18 patients. Five basic types of tumor-bronchus relationships were identified with CT: (a) bronchus cut off by the tumor, (b) bronchus contained within the tumor, (c) bronchus compressed by the tumor, (d) thickening and smooth narrowing of the bronchus leading to the tumor, and (e) thickening and irregular narrowing of the bronchus leading to the tumor. The diagnostic yield of transbronchial forceps biopsy and bronchial brushing was significantly higher in nodules characterized by a cut-off or contained bronchus. Transbronchial needle aspiration was performed in six patients, and results were positive in five, all of whom had a compressed or thickened bronchus. These results confirm that yield of transbronchial biopsy is determined by the type of tumor-bronchus relationship and the biopsy technique performed.