Foveal avascular zone segmentation using deep learning-driven image-level optimization and fundus photographs.

The foveal avascular zone (FAZ) is a retinal area devoid of capillaries and associated with multiple retinal pathologies and visual acuity. Optical Coherence Tomography Angiography (OCT-A) is a very effective means of visualizing retinal vascular and avascular areas, but its use remains limited to research settings due to its complex optics limiting availability. On the other hand, fundus photography is widely available and often adopted in population studies. In this work, we test the feasibility of estimating the FAZ from fundus photos using three different approaches. The first two approaches rely on pixel-level and image-level FAZ information to segment FAZ pixels and regress FAZ area, respectively. The third is a training mask-free pipeline combining saliency maps with an active contours approach to segment FAZ pixels while being trained on image-level measures of the FAZ areas. This enables training FAZ segmentation methods without manual alignment of fundus and OCT-A images, a time-consuming process, which limits the dataset that can be used for training. Segmentation methods trained on pixel-level labels and image-level labels had good agreement with masks from a human grader (respectively DICE of 0.45 and 0.4). Results indicate the feasibility of using fundus images as a proxy to estimate the FAZ when angiography data is not available.

Megapixel multi-elemental imaging by Laser-Induced Breakdown Spectroscopy, a technology with considerable potential for paleoclimate studies.

Paleoclimate studies play a crucial role in understanding past and future climates and their environmental impacts. Current methodologies for performing highly sensitive elemental analysis at micrometre spatial resolutions are restricted to the use of complex and/or not easily applied techniques, such as synchrotron radiation X-ray fluorescence micro-analysis (µ-SRXRF), nano secondary ion mass spectrometry (nano-SIMS) or laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Moreover, the analysis of large samples (>few cm²) with any of these methods remains very challenging due to their relatively low acquisition speed (~1-10 Hz), and because they must be operated in vacuum or controlled atmosphere. In this work, we proposed an imaging methodology based on laser-induced breakdown spectroscopy, to perform fast multi-elemental scanning of large geological samples with high performance in terms of sensitivity (ppm-level), lateral resolution (up to 10 µm) and operating speed (100 Hz). This method was successfully applied to obtain the first megapixel images of large geological samples and yielded new information, not accessible using other techniques. These results open a new perspective into the use of laser spectroscopy in a variety of geochemical applications.

Reconstructing a Carboniferous inferred coral-alcyonarian association using a biomineralogical approach.

The taxonomic assignation and ecological implications of the genus Syringoalcyon Termier & Termier, 1945 have been a palaeontological problem for a long time. Carboniferous material from Morocco and Spain has been studied using a biomineralogical approach by means of petrographic microscopy, SEM, AFM, EMPA and CIP microscopy analysis. Detailed morphological, structural, chemical composition and crystallographic data enable a deeper understanding of the nature of Syringoalcyon. The coral walls and the so-called epithecal scales exhibit conspicuous differences in microstructure (lamellae and holacanthine fibres in the coral vs. single crystal in scales), nanostructure (pill-shaped vs. granule-shaped nanocrystals), composition (LMC vs. HMC) and crystallographic orientation. The results of these analyses imply that Syringoalcyon is an association between the tabulate coral Syringopora and an epibiont. They also suggest that the epibiont was an alcyonarian (a rare occurrence in the fossil record) that was attached to the syringoporoid. This work highlights the utility of the biomineralizational approaches for solving palaeontological problems, such as systematic affinities, and for advancing knowledge of the evolution of biocrystallization processes.

Development of isolation-induced vocal behavior in normal-hearing and deafened guinea pig infants.

Infants in many different animal species require auditory information from conspecifics to learn appropriate responses to important environmental and social cues. Isolation calls are emitted by infant guinea pigs in contexts of social separation from their mothers. The aim of the present study was to examine the ontogeny of the isolation calls in normal-hearing and deafened infant guinea pigs, from 2 to 40 days postpartum and to determine the role of hearing maternal vocalization in infant guinea pig vocal responses in contexts of social proximity and isolation. Female newborn pigmented guinea pigs (Cavia porcellus) were housed with their birth mother and siblings. Water and dry food were available ad libitum. One day postpartum, the cochlea of infants in the experimental group was destroyed. The control group consisted of normal-hearing female siblings. Vocalizations from infants in the experimental and control groups were recorded for 6 minutes when with the mother before isolation, 6 minutes when alone, and then 6 minutes when with the mother after reunion. Recordings were made 5 days per week from 2 to 40 days after birth. The duration of calling was calculated for each 6-minute period of recording. Results demonstrated that deaf infants vocalize more than normal-hearing infants during social isolation from their mothers. Vocal activity of isolated deaf and normal-hearing infants decreased substantially over development, almost disappearing by the end of the study period. These results indicate that maternal vocal behavior modulates the vocal responses of guinea pigs early in infant development and supports other evidence that the guinea pig offers a viable model for investigating audition in deaf and normal-hearing human infants.

Gastric distension modulates hypothalamic neurons via a sympathetic afferent path through the mesencephalic periaqueductal gray.

The effects of gastric distension on extracellularly recorded single neuronal activity in the lateral hypothalamuslateral preoptic area-medial forebrain bundle (LPA-LH-MFB), other areas of the hypothalamus, mesencephalic periaqueductal gray (PAG), and other areas associated with the mesencephalic reticular formation were determined in the anesthetized rat. Gastric distension was produced by filling a gastric balloon with water using a calibrated infusion pump. Experimental conditions were based on previous studies that simulated gastric distension during fluid consumption in the rat. The effects of stomach distension using water at body temperature and room temperature were compared. Neurons in both the hypothalamus and mesencephalon were modulated by gastric distension. Hypothalamic neurons exhibited responses associated with gastric distension and exhibited interactions between distension and temperature stimulation of the stomach. Neurons in the mesencephalic periaqueductal gray (PAG) and associated reticular formation also were modulated by these gastric stimuli. When the PAG was electrically stimulated, similar responses to gastric distension and PAG stimulation were observed for hypothalamic neurons. The effects of gastric distension on hypothalamic neurons were reduced or eliminated when the PAG stimulating electrode site was destroyed by electrocoagulation. In addition, the microiontophoretic application of horseradish peroxidase at hypothalamic neuronal recording sites where gastric distension effects were observed resulted in the retrograde labeling of neurons in the PAG. These gastric stimulation-induced effects on hypothalamic and mesencephalic neuronal activity were attenuated but were not permanently eliminated by bilateral cervical vagotomy. However, these effects were significantly reduced or eliminated by bilateral transection of the cervical sympathetic chain or spinal transection at the first cervical level. Because the filling of balloons placed into the abdominal cavity close to the stomach had no similar effects on neural activity, these results can be attributed primarily to the activation of gastric mechano-and temperature-sensitive receptors. These results indicate that the effects of gastric temperature/distension stimulation under these conditions are mediated to a large degree by sympathetic afferents. The PAG is clearly involved as one of the mesencephalic relays for gastric afferent input to the hypothalamus.

Effects of neurotransmitters and vagus nerve stimulation on diencephalic and mesencephalic neuronal activity.

The effects of microiontophoretically applied neurotransmitters and cervical vagus nerve stimulation on neuronal discharge frequency was determined for cells located in the lateral hypothalamus-lateral preoptic area (LH-LPA), medial hypothalamus, thalamus-zona incerta area, and midbrain of anesthetized rats. Seven barrel electrodes were utilized to record simultaneously from and apply gamma aminobutyric acid (GABA), dopamine, glutamate, acetylcholine and norepinephrine to single neurons. Microiontophoretically applied GABA resulted in similar decreases in neuronal discharge frequency in all central areas tested. Thalamus-zona incerta cells were the most sensitive to GABA and required the lowest ejection current for threshold responses. Dopamine application resulted in increases and decreases in hypothalamic and mesencephalic neuronal discharge frequency. Thalamus-zona incerta neurons were the most sensitive to dopamine and exhibited only decreased activity during its application. Glutamate application resulted in a non-specific excitation of neurons. Acetylcholine induced increases and decreases in neuronal discharge frequency. Thalamus-zona incerta cells were the most sensitive to acetylcholine application. Hypothalamic neurons were the most sensitive to norepinephrine induced decreases in neuronal activity and effects in this area persisted for a prolonged period after norepinephrine was applied. Neuronal discharge frequency was significantly increased during vagus nerve stimulation in all central areas studied. Only hypothalamic neurons also exhibited decreased activity during vagus nerve stimulation. Results are discussed in terms of previous neurochemical and neurophysiological data and in terms of the importance of vagal afferents in the control of central neuronal activity.

Caudate unit activity and somal diameters in intact and nigral lesioned cats.

In a search for morphofunctional relationships in the head of the caudate nucleus (CN), we recorded extracellular unit activity in intact cats and in cats that had received bilateral injections of 6-OHDA into the substantia nigra (SN) 30 days previously. Only units firing spontaneously and continuously for 2 min were studied. In dorsal regions, potentials were small and iterative at almost constant intervals; the somal diameters were relatively small. In the ventrolateral region, potentials were bigger and appeared in bursts; somal diameters were significantly larger (p less than 0.05). For the centromedial region a histogram of numbers of neurons as a function of diameters revealed a Gaussian distribution extending from small to large neurons. Most dorsal neurons increased their firing rate to radial nerve, visual, SN, and/or nucleus centralis medialis (NCM) stimulation. Ventral neurons usually responded with excitation followed by long lasting inhibition, particularly to SN and NCM stimulation. A few neurons responded to all four inputs and some showed long-lasting potentiation in response to low frequency stimulation, suggesting a more general function. Greatest convergence (65%) was found for NCM and SN inputs. In lesioned cats, there was no SN driving, NCM's inhibitory actions almost disappeared, and the excitatory action of the other stimuli was reduced.

Mesencephalic reticular formation stimulation effects on hypothalamic neuronal activity.

The effects of mesencephalic reticular formation (RF) single pulse, 0.5 msec and 0-500 microA, stimulation on lateral preoptic-lateral hypothalamic (LPA-LH) neuronal activity were determined in anesthetized rats. In addition, the effects of LH stimulation on neural activity in the RF and periaqueductal gray (PAG) were evaluated. Recordings from 117 neurons indicate reciprocal connections between the LPA-LH and the mesencephalon. Stimulation of the RF affected 70% of the LPA-LH neurons tested. Short latency decreases in activity predominated indicating an inhibitory synaptic input from the RF to the LPA-LH. Short latency increases in discharge frequency were observed infrequently. Stimulation of the LH affected only 32% of the mesencephalic neurons tested. Short latency decreases in activity were usually observed indicating reciprocal inhibitory synaptic connections between the LPA-LH and the RF and periaqueductal gray. Antidromic responses verified these interconnections and revealed relatively slow conduction velocities of approximately 1.0 m/sec. Results are discussed in terms of the involvement of the LPA-LH and RF in sensorimotor functions, spinal motor excitability, and ingestive behavior.

Effects of ventral tegmental area stimulation and microiontophoretic application of dopamine and norepinephrine on hypothalamic neurons.

The effects of ventral tegmental area of Tsai (VTA) stimulation on lateral hypothalamic (LH), lateral preoptic area (LPA). and medial hypothalamic neuronal activity were determined in anesthetized rats. Recordings from 81 hypothalamic neurons indicate that stimulation produces predominantly decreases in hypothalamic neuron activity. Increase in activity due to VTA stimulation occurred less frequently. Following single rectangular pulse stimulation, 0.5 msec. 0-500 microA, short latency decreases in activity occurred. Longer latency increases in discharge frequency were also observed. Dose response relations were established for 56% of the LH neurons, 78% of the LPA neurons, and for 82% of the medial hypothalamic neurons following VTA stimulation. Decreases and in a few cases increases in activity seemed to involve only one or two synapses. Antidromic responses verified interconnections between the VTA and the hypothalamus and revealed relatively slow conduction velocities of 0.45 and 0.81 m/sec. The changes in discharge frequency which occurred following VTA stimulation were similar in direction to the effects of the direct microiontophoretic application of dopamine (DA) or norepinephrine (NE). Since DA increased or decreased while NE decreased discharge frequency, these microiontophoretic tests indicated that the shorter latency VTA stimulation induced increases in decreases in neural activity were associated with VTA dopaminergic neuron stimulation and that in some cases short and long latency decreases in neuronal activity were due to activation of VTA ventral bundle NE fibers of passage or to indirect polysynaptic mechanisms. Results demonstrate the interconnections between various regions of the hypothalamus and the VTA along the extent of the medial forebrain bundle (MFB). The cross-validation of neuroanatomical and various electrophysiological methods in establishing the nature of hypothalamic connections was discussed.

Effects of periaqueductal gray stimulation of diencephalic neural activity.

The effects of ipsilateral mesencephalic periaqueductal gray (PAG) stimulation on lateral hypothalamic (LH), lateral preoptic area (LPA), and ventral and subthalamic activity were determined in anesthetized rats. Recordings from 119 diencephalic neurons indicate that the PAG provides a predominantly inhibitory input to diencephalic neurons. Excitatory input occurred infrequently in the hypothalamus and was not observed in the thalamus. Following single rectangular pulse stimulation, 0.5 msec, 0-500 micro A, short latency decreases in activity occurred. Longer latency increases in discharge frequency were also observed. Dose response relations were established for 74% of the LH neurons, 68% of the LPA neurons, and for 72% of the ventral and subthalamic neurons following VTA stimulation. Decreases and, in a few hypothalamic neurons, increases in activity seemed to involve only one or two synapses. The effects of contralateral PAG stimulation on LPA-LH neuronal activity were alos determined. Dose response relations were established for 66% of the LPA-LH neurons following contralateral stimulation. However, results were different in that many more cells were increased with a shorter latency and at a lower threshold following contralateral stimulation. Antidromic responses verified PAG and diencephalic interconnections and revealed relatively slow conduction velocities, less than 1.0 m/sec. Results were discussed in terms of the anatomy of known PAG pathways, PAG neuronal activation vs. PAG fibers of passage, and the functions of midbrain-hypothalamic interconnections in the integration of somatic, visceral and nociceptive sensory inputs.

Facilitation of conditioned motor suppression by microinjections of dopamine in the caudate nucleus of cats.

The effects of Caudate Nucleus (CN) injections of catecholamines on the suppression of a motor conditioned response (MCR), lever pressing, was investigated. Cats were trained to press a lever to obtain 0.5 ml of milk when a conditioned discriminative stimulus was on (CS-on-reward on, MCR) and to suppress the response when the light was off (CS-off-reward off, suppression of motor conditioned response, SMCR). The bilateral application of 5 or 10 micrograms of Dopamine (DA) through chronically implanted cannulae in the CN significantly decreased the lever pressing in the non rewarding situation without changing the MCR. Injection of 5 micrograms of L-DOPA caused very small effects assessed during the following 10 min. However, 10 micrograms of L-DOPA produced a significant decrement of lever pressing (CS-off) in two out of four injections. These findings further support the postulation that catecholamines in the CN have a behavioral inhibitory action upon a motor conditioned response.