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Fine-Scale Landscape Genomics Show Asymmetric Patterns of Gene Flow for the Invasive Mosquito Aedes albopictus.

Mosquito-borne viruses like dengue, Zika, and chikungunya pose increasing health risks in the United States due to the expanding range of Aedes albopictus, a highly invasive mosquito species that now has a global distribution. Aedes albopictus thrive in artificial containers associated with anthropogenic land use, allowing populations to reach high numbers in urban and suburban environments. While the global spread of Ae. albopictus has been well characterized, the effects of heterogeneous urban landscapes on dispersal and gene flow at fine spatial scales remain unclear. This study analyzed the genetic connectivity of Aedes albopictus populations collected in Wake County, North Carolina in 2018. We used single nucleotide polymorphisms (SNP) data from double-digest restriction-enzyme associated DNA sequencing (ddRADseq) and examined genetic connectivity through principal component analysis (PCA) and genetic network analysis. We then evaluated migration and source-sink dynamics using a Bayesian approach for SNP data (BA3-SNP). We found little evidence of genetic clustering or isolated populations of Ae. albopictus in Wake County, suggesting high gene flow between sites. Migration analysis demonstrated asymmetric gene flow from rural to urban regions within Wake County, with greater gene flow occurring between and within urban regions. These findings suggest that the pattern of gene flow of Ae. albopictus populations within local metropolitan areas may involve urban city centers serving as genetic sinks and surrounding suburban and rural regions serving as sources. This study highlights how heterogeneous landscapes shape mosquito population connectivity and migration at fine spatial scales, which is critical for informing vector control and public health intervention strategies.

Aedes albopictus

Quantitative Fluorescence Imaging of Alphavirus Infection for Antiviral Screenings.

Fluorescence microscopy offers a highly sensitive and versatile approach for investigating alphavirus infection at the cellular level. By combining fluorescently labeled viruses with quantitative image analysis, this method enables detailed spatial and temporal characterization of infection dynamics, including the detection of subtle differences in replication kinetics and cell-to-cell spread. A central aim of this protocol is its application in antiviral screening assays. Image-based quantification of fluorescence intensity provides a robust and reproducible means to assess the efficacy of antiviral compounds, allowing early and sensitive detection of inhibitory effects in infected cells. This facilitates the identification of promising antiviral hits and supports the evaluation of dose-dependent responses. The approach is also well-suited for comparative studies of different alphavirus strains or mutants, as variations in replication behavior and dissemination patterns become readily apparent. Its flexibility, compatibility with multiple cell lines, and straightforward integration into automated imaging platforms makes the method scalable and suitable for high-throughput screening campaigns. Overall, this protocol advances the discovery and evaluation of antiviral strategies. Given that several alphaviruses cause significant human and veterinary diseases, lack approved antiviral therapies, and continue to expand geographically with emerging outbreaks, the identification of novel antivirals remains an urgent priority. Therefore, this fluorescence-based workflow represents a valuable and timely contribution to modern alphavirus research.

Antiviral Agents

Genomic signatures of adaptation across a landscape of crickets following the introduction of a parasitoid fly.

Novel species interactions provide an opportunity to assess the earliest stages of genetic adaptation. We combined population genomics and field selection surveys to explore the genomic and geographic landscape of adaptation in small, fragmented Hawaiian cricket populations, which are parasitized by larvae of an introduced fly that targets singing males. Multiple protective male-silencing cricket morphs have recently spread under this novel selection pressure, despite song's important roles in mate attraction. We find evidence of sharp declines in cricket effective population sizes following the fly's introduction and identify regions under spatially varying selection mediated by infestation risk, which are linked to an adaptive morph and other genes implicating in resisting infestation. Despite repeated bottlenecks, genetic variation is dominated by structural variant polymorphisms seemingly maintained under balancing selection. Our study illustrates pervasive consequences of abrupt changes in selection on small populations. Fly-mediated selection remains strong despite the spread of adaptive male-silencing phenotypes, which also reduce male fitness in the context of mate attraction.

Animals

Spatial resolution and count density requirements in brain SPECT imaging.

A set of simulations has been performed to investigate the spatial resolution and count density requirements for brain SPECT imaging. Projections were drawn from a matrix representation of the Hoffman brain phantom. These projections were convolved with realistic point spread functions and Poisson noise was added to simulate a wide range of imaging situations normalized to a fixed imaging time. The projections were optimally smoothed with a Wiener filter and were reconstructed with a ramp filter. The quality of the reconstructed images was determined objectively from the normalized mean square between the simulated data and the true distribution. This ranking was validated against the preferences of a group of trained observers. The results from this study indicate that the optimal choice of spatial resolution (collimation) depends on the available count density. As the count density (normalized to 10 mm resolution) increases by a factor of 2.7, results from the simulations indicate that the optimal spatial resolution improves by 1 mm. For brain studies in which the administered activity is limited (such as 123I IMP), the optimal spatial resolution is approximately 8 to 9 mm. With 99Tcm labelled brain agents the amount of administered radioactivity can be increased six-fold and the optimal spatial resolution is predicted to fall to about 6 to 7 mm. If sensitivity is further increased by the use of a dedicated SPECT unit with multiple detectors, the optimal spatial resolution will be on the order of 4 to 5 mm.

Brain

Calcium transients evoked by climbing fiber and parallel fiber synaptic inputs in guinea pig cerebellar Purkinje neurons.

1. Calcium transients related to climbing fiber (CF) and parallel fiber (PF) synaptic potentials were recorded from Purkinje cells in guinea pig cerebellar slices. Transients were measured using either absorbance changes of arsenazo III or fluorescence changes of fura-2, which were injected into individual cells in the slice. 2. All-or-none somatically recorded CF potentials elicited by white matter stimulation had all-or-none Ca transients. These signals began with a delay of > or = 2 ms from the start of the electrically recorded synaptic potential. The recovery time of CF-induced arsenazo III absorbance transients was < 50 ms in the fine dendrites in conditions that minimized the effects of dye buffering. 3. Ca2+ entry through voltage-gated Ca channels opened by Ca action potentials was the dominant source of the rise in [Ca2+]i after CF activation. There was no significant change in [Ca2+]i corresponding to the plateau potential that followed the large CF response. 4. The appearance and amplitude of distal CF-evoked Ca signals was more variable than proximal signals, suggesting that CF potentials do not reliably spread to the fine distal dendrites. The distal transient could be enhanced by intrasomatic depolarizing pulses, suggesting that it was a property of the postsynaptic membrane and not the presynaptic side of the CF synapse that was responsible for this variability. 5. Parallel fiber responses were evoked by electrical stimulation near the pial surface. Graded synaptic potentials and related Ca transients were reversibly blocked by 2 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Small synaptic potentials induced small, localized Ca transients. With increasing stimulus intensity, the PF electrical response developed a regenerative component. Larger dendritic Ca transients were detected corresponding to this component. Ca transients evoked by the regenerative responses had the same rapid rise times and fall times as those related to somatically stimulated Ca action potentials, suggesting that they also were due to Ca2+ entry through voltage-sensitive channels. 6. During trains of PF responses, we observed an increase in the spatial extent of related Ca transients. This effect could be modulated by changes in the resting potential, suggesting that the same intrinsic mechanism was affecting the spread of both CF and PF signals.

6-Cyano-7-nitroquinoxaline-2,3-dione

Membrane currents in the rabbit sinoatrial node cell as studied by the double microelectrode method.

When a strand of the rabbit sinoatrial node tissue was shortened by ligation, the spatial decay of electrotonic potential decreased and the input impedance increased. In a piece of the tissue 0.2-0.3 mm in diameter apparently uniform current spread was obtained. Action potentials recorded from three different sites in this small piece occurred simultaneously and were superimposable. In voltage clamp experiments using the double microelectrode method, the membrane potential was usually held at -30 to -40 mV, where no net current flowed. When membrane potential was suddenly changed from the holding potential, the sign and the time course of the ionic current varied with membrane potential. Hyperpolarization gave an inward current which increased with time. Depolarization gave a transient inward current followed by sustained outward current, and repolarization gave an outward current tail which exponentially subsided with a time constant of 0.37 s. The membrane time constant was 12.0 ms. When the specific membrane capacitance was assumed to be 1 muF/cm2, the specific membrane resistance at the resting potential was 12 Komega cm2. The peak of the transient inward current on depolarization was 1.3 X 10(-5) A/cm2.

Action Potentials

[Structure of the morphogenetic movements of gastrulation in Anura. II. The elementary morphogenetic processes].

The clinostate rotation causes a wide range of gastrulation abnormalities in the Anura. A special investigation has shown that all gastrulation variants may be represented as various spatial-temporal compositions of the same morphogenetic processes isolated by means of formal experimental procedure. These processes are structurally stable rearrangements of tissue morphology and are spread in the tissue as a wave embracing more and more cells. The change of one local morphology for another may proceed either in the form of "shock wave", a moving distinct border between the cells already switched and not yet switched to the new automorphous movement, or in the form of "deployment" when the transition from one local morphology to another is continuously deployed in space.

Amphibians

Cerebral blood flow in the monkey after focal cryogenic injury.

A focal cryogenic lesion was made in the left superior frontal gyrus of the anesthetized macaque brain. Cerebral blood flow (CBF) was determined by the hydrogen clearance technique before and during the 4 hours following trauma. Local CBF in tissue adjacent to the lesion increased in the first half hour after the lesion was made and then decreased during the ensuing 3 1/2 hours. Local CBF in the contralateral superior frontal gyrus, as well as total CBF and oxygen consumption, were unchanged by cryogenic trauma. The spread of vasogenic edema into uninjured tissue probably accounts for the observed decrease in local CBF. This experimental model may assist in discovering therapy to alter favorably the spatial and temporal profile of pathologic CBF changes in tissue surrounding an acute lesion of the brain.

Animals

Modeling of the quantal release at interneuronal synapses: analysis of permissible values of model moments.

A theoretical study of effects of the different factors on fluctuation of post-synaptic potential (PSP) amplitudes was undertaken, using computation of regions of permissible values (RPV) of the ratio between the variance and the mean number of the quanta released (R1) and the ratio between the third moment and the variance (R2). The RPVs of these indexes for the binomial model were compared with regions determined for a number of models incorporating several factors. It has been shown that the involvement of temporal non-uniformity of transmitter release probability, decremental spreading of potentials along dendrites, and failure of spike propagation give the values of skewness index R2 less, compared to the binomial model. Simultaneously, a number of other factors, especially spatial non-uniformity of release probabilities in single release sites, would give amplitude histograms with high positive values of the index. The values of R1 and R2, calculated for 21 samples of sensorimotor EPSP amplitudes, were biased from RPV of these parameters constructed for the binomial model. The scattergram of R1 and R2 can be explained by the presence of two kinds of contacts which release quantum with different probabilities. The same was true for the beta-model based on the assumption that probabilities of quantal release are a sample of values of random variable that has beta-distribution. From analysis of the distribution of individual release probabilities, obtained from evaluation of beta-model parameters, is concluded that a greater part of boutons in the sensorimotor synapses release transmitter with very low probabilities, there being, however, a few boutons with probabilities close to 1.

Animals

Polarized infrared spectroscopy of oriented purple membrane.

Polarized Fourier transform infrared spectroscopy has been used to study the structure of purple membrane from Halobacterium halobium. Membranes were oriented by drying a suspension of membrane fragments onto Irtran-4 slides. Dichroism measurements of the amide I, II and A peaks were used to find the average spatial orientation of the bacteriorhodopsin alpha-helices. By deriving a function that relates the observed dichroism to the orientational order parameters for the peptide groups, helical axis distribution, and mosaic spread of the membranes, the average orientation of the alpha-helices was found to lie in a range of less than 26 degrees away from the membrane normal, agreeing with electron microscopic measurements. The frequency of the amide I and A peaks is at least 10 cm-1 higher than values found for most alpha-helical polypeptides and proteins. This may indicate that bacteriorhodopsin contains distorted alpha-helical conformations.

Bacteriorhodopsins

The line spread function and modulation transfer function of a computed tomographic scanner.

A simple method to measure the line spread function of computed tomographic scanners has been developed. The line spread function of the EMI-Scanner was measured and the MTF of the scanner was calculated. The reproducibility of the method indicated this technique could be utilized to compare the spatial resolution of various computed tomographic scanners.

Diagnosis, Computer-Assisted

Temperature topography of the brain cortex: thermoencephaloscopy.

Thermoencephaloscopy (TES) - a new method of functional imaging of the cerebral cortex by its infrared radiation was advanced and developed since 1984. Improved thermovision and image processing techniques allow 2D, contact-free, dynamic and non-invasive recording of background and evoked cortical activity through an unopened skull. Activated (heated) and deactivated (cooled) zones of the cerebral cortex are revealed. The temporal resolution of TES is 40 msec (25 maps/sec), the spatial resolution is up to 70 x 70 microns/pixel. The diameter of the smallest recordable active region of the cortex is 200-300 microns. The minimal time needed for a session used for averaging of 4-9 responses varied from 40 sec up to 18 min. TES allows to detect the position, size and sequence of operation of precisely located specific cortical zones, and to measure their dynamics before, during and after sensory and direct cortical stimulation, motor acts and conditioning (associative learning). TES-effects were recorded in rats, rabbits, cats, monkeys and humans. Waves were found spreading over the cortex with a speed of up to 30 mm/sec along trajectories specific for the sensory modality and the site of stimulation. Some pathological processes in the brain are detectable by TES: experimental tumours and epileptic foci. There are many sources for local heating: neural activity, local metabolism of units, local cerebral blood flow and thermoconductivity in the activated zones of the cortex. Thermoencephaloscopy is a dynamic, non-invasive, contact-free method with a relatively high temporal and spatial resolution and sensitivity. It can be a useful tool in basic neuroscience and medicine.

Acoustic Stimulation

Localized desensitization of Limulus photoreceptors produced by light or intracellular calcium ion injection.

Spots of light were used to measure the light sensitivity of spatially separated regions of single Limulus photoreceptors. The desensitization caused by irradiating part of the cell was largest in the irradiated region. The desensitization caused by intracellular calcium ion injection was largest near the infection site. The spread of desensitization away from the injection site suggests that calcium ion can diffuse over neuronal dimensions, but that the effective rate of diffusion is not so high as to abolish calcium gradients. The results are compatible with the previously proposed hypothesis that a rise in the intracellular calcium ion concentration mediates light adaptation.

Animals

The effects of selective glutamate receptor antagonists on synchronized firing bursts in layer III of rat visual cortex.

In the rat visual cortex in vitro, single-shock stimulations applied to the border between layer VI and the white matter evoke synchronized burst-firing by units in layer III. We have examined the effects of glutamate receptor antagonists on this activity, with antagonists applied via the bath to allow correlation of effects with concentrations. All synaptically driven components (recorded extracellularly as field potential 'S2' spikes, dipoles 'W1' and 'W2', and coinciding single-unit spikes) were inhibited by greater than 90% in 1.0 mM kynurenic acid and in 3 or 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, which selectively blocks AMPA/kainate receptors). S2 spike amplitudes were reduced by half in 0.7 microM CNQX. 2-Amino-5-phosphonovalerate (APV), a specific blocker of NMDA receptors, did not prevent S2 spike burst or horizontal spread of bursting within layer III. However, APV reduced the duration of synchronized bursts and the slower potentials which followed. In Mg(2+)-free medium, new components appeared which were APV-sensitive: (1) low amplitude spikes, distributed spatially like S2 spike, but recurring more slowly, and (2) slow potentials, distributed spatially like W1 and W2 potentials, but lasting for hundreds of milliseconds. The amplitudes of these spikes were reduced by half in 3 microM D-APV. Our data imply that: (1) glutamate receptors play a major role in mediating local, excitatory neurotransmission in the supragranular layers of neocortex, with NMDA and AMPA/kainate subtypes each subserving somewhat different functions; (2) AMPA/kainate receptors mediate rapid excitatory transmission between layer III neurons, responsible for driving the first 15 ms of synchronized bursts; (3) currents gated by NMDA receptors determine the duration of coherent firing bursts, and drive asynchronous neuronal firing following bursts; and (4) under conditions which circumvent block by extracellular Mg2+, activation of NMDA receptors greatly enhances and prolongs the response to single-shock stimulations. In vivo, activation of layer III neurons is likely to depend significantly upon currents gated by NMDA receptors whenever repetitively firing excitatory inputs summed over several tens of milliseconds provide enough depolarization to lift block by extracellular Mg2+.

2-Amino-5-phosphonovalerate

Detection of cortical spreading depression in the anaesthetised rat by impedance measurement with scalp electrodes: implications for non-invasive imaging of the brain with electrical impedance tomography.

The possible use of impedance measurement with scalp electrodes to detect intracranial events non-invasively was investigated by measuring the localised impedance changes during cortical spreading depression (CSD) in anaesthetised rats. Impedance was measured over ipsilateral fronto-parietal cortex by a four electrode method operating at 50 kHz with electrodes spaced 0.5 mm apart. Cortical impedance increased by 39% of the resting level during CSD. With scalp electrodes placed on abraded skin, an unexpected impedance decrease of 0.8% occurred, which correlated spatially and temporally with CSD. CSD was accompanied by a small rise in temperature; when scalp temperature was held constant by warming the scalp, no impedance change greater than baseline variability (+/- 0.1% of the resting impedance level) was observed. The non-invasive detection of CSD in migraine in humans may be possible by measuring the characteristic temperature-related impedance changes with Electrical Impedance Tomography and scalp electrodes.

Anesthesia, Inhalation

Multiple excitations in photosynthetic systems.

The yield of fluorescence in Chlorella from a 7 ns pulse of light is found to decrease gradually as a function of the number of hits in the photosynthetic units. The fivefold decrease in yield is spread over some three orders of magnitude of pulse energy and strongly suggests another random process in addition to that of photon absorption. Evidence supports the view that this random process is not in the time but in the spatial domain. The model used to fit the data is that of a unit with multiple traps for the singlet excitation. An excitation is captured by an open trap or destroyed by a filled trap with equal probability. These studies give evidence for the connectivity of the photosynthetic energy transfer apparatus on the short time scale. The short fluorescence lifetimes following picosecond pulse excitation of photosynthetic systems reported by several laboratories may be explained by the effect of multiple excitations.

Chlorella

Signal and noise in modulation transfer function determinations using the slit, wire, and edge techniques.

The modulation transfer function (MTF) of an idealized imaging system can be determined from the Fourier transform of the system's line-spread function (LSF). Three techniques of experimentally determining the LSF require imaging either a slit, wire, or edge. In this paper, these three techniques are modeled theoretically to determine the noise in the calculated MTFs as a function of spatial frequency resulting from both quantum fluctuations and stochastic detector noise. The techniques are compared using the signal-to-noise ratio (SNR) in the MTF, defined as the ratio of the MTF value to the standard deviation in an ensemble of MTF determinations from independent measurements. It is shown that for a specified photon fluence, the edge method MTF has the highest SNR at low spatial frequencies, while that of the slit method is superior at high frequencies. The wire method SNR is always inferior to that of the slit technique. This suggests that the edge method is preferable for measuring parameters such as the low-frequency drop, and the slit method is preferable for determining high-frequency response. The cross-over frequency at which the slit and edge methods are equal (f(e)) for quantum-noise limited systems is a function of the slit width and the length over which the LSF is measured. For detector-noise limited systems, f(e) is dependent on the slit width only. The SNR in all but the quantum-noise limited slit method can therefore be increased by decreasing the length over which the LSF is measured, smoothing the tails of the LSF, or by fitting the tails to an analytic expression.

Fourier Analysis

[Spatial synchronization of rabbit brain biopotentials upon repeated electrical stimulation of the cortex].

A study was made on 23 rabbits of the degree of spatial synchronization in response to repeated electrical stimulations of one or simultaneously two cortical areas: the sensorimotor or visual, or simultaneously the sensorimotor and visual, or sensorimotor and precentral respectively. Enhancement of the synchronization induced by the stimulation, globally spread over the entire cortex and was followed by its decrease. However, in the two directly stimulated areas of the cortex, the similarity of bioelectrical oscillations persisted for a long time, which testifies to the establishment of selective relations between them.

Animals