A simple multidimensional system for the recording and interpretation of sperm morphology in bulls

This study reports on the development of a data capture sheet for recording the sperm morphology of bulls. The arrangement of the data capture sheet permits the simultaneous recording of the viability and morphology of spermatozoa, and the recording of multiple defects in spermatozoa. Apart from deriving the prevalence of each type of defect, the sheet permits the derivation of the percentages spermatozoa with nuclear defects, defects of the acrosome or tail, respectively, as well as the percentages morphologically normal and normal viable spermatozoa. The arrangement of the data capture sheet further permits the rapid and easy interpretation of the sperm morphology regarding the stage of development, site and extent of malfunction of the reproductive organs, the prognosis for change in the sperm morphology over time and the extent to which defects may be compensated for by normal spermatozoa and the likely fertility of the particular semen sample. The data capture sheet permits a multidimensional, holistic interpretation of the sperm morphology of a bovine semen sample.     

Designing a nanotube using naturally occurring protein building blocks

Here our goal is to carry out nanotube design using naturally occurring protein building blocks. Inspection of the protein structural database reveals the richness of the conformations of proteins, their parts, and their chemistry. Given target functional protein nanotube geometry, our strategy involves scanning a library of candidate building blocks, combinatorially assembling them into the shape and testing its stability. Since self-assembly takes place on time scales not affordable for computations, here we propose a strategy for the very first step in protein nanotube design: we map the candidate building blocks onto a planar sheet and wrap the sheet around a cylinder with the target dimensions. We provide examples of three nanotubes, two peptide and one protein, in atomistic model detail for which there are experimental data. The nanotube models can be used to verify a nanostructure observed by low-resolution experiments, and to study the mechanism of tube formation.     

组织工程细胞片的构建

作为一种无支架的组织工程技术,组织工程细胞片不仅可以避免支架材料带来的不利影响,而且可通过组装进一步形成更为复杂的三维功能化组织,因而在生物医学领域备受关注.细胞片的构建主要是基于敏感性材料所构建的培养基底,通过改变温度、酶、光、离子、氧化还原pH、糖等刺激因素,调节基底对细胞的粘附行为使细胞发生自然脱附,从而获取细胞...     

Development of a bio-MEMS device for electrical and mechanical conditioning and characterization of cell sheets for myocardial repair

Here we propose a bio-MEMS device designed to evaluate contractile force and conduction velocity of cell sheets in response to mechanical and electrical stimulation of the cell source as it grows to form a cellular sheet. Moreover, the design allows for the incorporation of patient-specific data and cell sources. An optimized device would allow cell sheets to be cultured, characterized, and conditioned to be compatible with a specific patient's cardiac environment in vitro, before implantation. This design draws upon existing methods in the literature but makes an important advance by combining the mechanical and electrical stimulation into a single system for optimized cell sheet growth. The device has been designed to achieve cellular alignment, electrical stimulation, mechanical stimulation, conduction velocity readout, contraction force readout, and eventually cell sheet release. The platform is a set of comb electrical contacts consisting of three-dimensional walls made of polydimethylsiloxane and coated with electrically conductive metals on the tops of the walls. Not only do the walls serve as a method for stimulating cells that are attached to the top, but their geometry is tailored such that they are flexible enough to be bent by the cells and used to measure force. The platform can be stretched via a linear actuator setup, allowing for simultaneous electrical and mechanical stimulation that can be derived from patient-specific clinical data.     

Evidence of clonal budding in a radial cluster of Paraconularia crustula (White) (Pennsylvanian: ?Cnidaria)

Monospecific conulariid clusters have been interpreted as products of gregarious behavior or as clonal colonies, with members of a colony originally inclined at a high angle to the substrate and connected to each other at their apices. However. individuals in all previously documented clusters are preserved parallel, or nearly parallel, to bedding, and their apices are not preserved. We describe a cluster of three Paraconularia crustula (White) (Pennsylvanian of Missouri, USA) in which the individual specimens exhibit an original steeply inclined orientation. The individuals in this cluster converge adapically on an extremely thin. crumpled sheet of apatitic material that is similar to conulariid test material. The conulariids and associated sheet bear a striking resemblance to regenerated, clonal colonies of polypoid coronatid scyphozoans. Such colonies arise from a thin, periderm-covered sheet of non-differentiated soft tissue, produced following severance of a single parent polyp. We suggest that the conulariid cluster originated through a similar process. Should this interpretation be corroborated by the discovery of better preserved conulariid clusters, it would provide strong additional evidence for the hypothesis of a scyphoroan affinity for conulariids     

Beta-sheet coil transitions in a simple polypeptide model.

A simplified model of a polypeptide chain is used to study the dynamics of the beta-sheet-coil transition. Each amino acid residue is treated as a single quasiparticle in an effective potential that approximates the potential of mean force in solution. The model is used to study the equilibrium and dynamic aspects of the sheet-coil transition. Systems studied include ones with both strands free to move (two-strand sheet), and ones with either strand fixed in position (multistrand sheet). The equilibrium properties examined include sheet-coil equilibrium constants and their dependence on chain position. Dynamic properties are investigated by a stochastic simulation of the Brownian motion of the chain in its solvent surroundings. Time histories of the dihedral angles and residue-residue cross-strand distances are used to study the behavior of the sheet structure. Auto- and cross-correlation functions are calculated from the time histories with relaxation times of tens to hundreds of picoseconds. Sheet-coil rate constants of tens of ns-1 were found for the fixed strand cases.     

Bring your own camera to the trap: An inexpensive,versatile, and portable triggering system tested on wild hummingbirds

The study of animals in the wild offers opportunities to collect relevant information on their natural behavior and abilities to perform ecologically relevant tasks. However, it also poses challenges such as accounting for observer effects, human sensory limitations, and the time intensiveness of this type of research. To meet these challenges, field biologists have deployed camera traps to remotely record animal behavior in the wild. Despite their ubiquity in research, many commercial camera traps have limitations, and the species and behavior of interest may present unique challenges. For example, no camera traps support high‐speed video recording. We present a new and inexpensive camera trap system that increases versatility by separating the camera from the triggering mechanism. Our system design can pair with virtually any camera and allows for independent positioning of a variety of sensors, all while being low‐cost, lightweight, weatherproof, and energy efficient. By using our specialized trigger and customized sensor configurations, many limitations of commercial camera traps can be overcome. We use this system to study hummingbird feeding behavior using high‐speed video cameras to capture fast movements and multiple sensors placed away from the camera to detect small body sizes. While designed for hummingbirds, our application can be extended to any system where specialized camera or sensor features are required, or commercial camera traps are cost‐prohibitive, allowing camera trap use in more research avenues and by more researchers.     

Microfabricated nerve–electrode interfaces in neural prosthetics and neural engineering

Neural interfaces and implants are finding more clinical applications and there are rapid technological advances for more efficient and safe design, fabrication and materials to establish high-fidelity neural interfaces. In this review paper, we highlight new developments of the microfabricated electrodes and substrates with regard to the design, materials, fabrication and their clinical applications. There is a noticeable trend towards integration of microfluidic modules on a single neural platform. In addition to the microelectrodes for neural recording and stimulation, microfluidic channels are integrated into a nerve–electrode interface to explore the rich neurochemistry present at the neural interface and exploit it for enhanced electrochemical stimulation and recording of the central and peripheral nervous system.     

Large-scale recording of neurons by movable silicon probes in behaving rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design. Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (http://www.neuronexustech.com/; http://www.sbmicrosystems.com/; http://www.acreo.se/) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.     

Comparison of audio recording system performance for detecting and monitoring songbirds

ABSTRACT Acoustic recording systems are being used more frequently to estimate habitat occupancy or relative abundance, and to monitor population trends over time. A potential concern with digital recording systems is that changes in technology could affect detectability of birds and cause bias in trend estimates based on counts of birds detected. We evaluated several currently available commercial recording systems ranging from low‐cost multipurpose digital recorders to custom‐designed wildlife recorders (US$250–$7000 price range) to examine possible differences among systems in species detection. We made recordings during Breeding Bird Surveys (BBS) counts using several units concurrently, and asked several expert birders to listen to the recordings in a factorial design. We found that birders detected, on average, 10% fewer species on some units compared to others, though there was high variance. Analysis of a subset of recordings, using spectrograms and repeated listening, suggested that ～90% of species on each BBS stop could be clearly detected on all units. The remaining species could be identified on at least one unit, but were hard or impossible to detect on others. We found that the recording unit with the lowest empirical signal‐to‐noise‐ratio (SNR) had the lowest number of birds detected on the BBS recordings, and that frequency‐specific SNR differed among units. Missed detections were likely related to variation in internal noise and frequency‐dependent sensitivity of the units, and were an issue for all systems regardless of price. We caution that researchers using recorders need to consider variation among recording systems in their study design, particularly for long‐term monitoring programs.     

VBA开发动物行为实时记录系统及实例

对动物行为进行实时记录是行为学研究的一项重要工作。传统的行为观察方法常常无法满足实时精确记录行为的要求 ,而一些现代的行为记录系统虽功能强大但却难以推广。作者应用VBA开发了一个简捷方便的行为实时记录系统 ,可以减少记录动物行为观察的工作量 ,降低记录动物行为的观察误差 ,保证了实验数据的可靠性。同时 ,该系统具有开放性和灵活性的特点 ,可以在不同动物行为研究中应用。     

New technique for studying reaction forces during primate behaviors on vertical substrates

Recording reaction forces from primates during behaviors on vertical substrates, such as leaping, climbing, or biting trees, typically requires the design and construction of customized recording devices or mounting commercially available force platforms in a vertical position. The technical difficulties imposed by either option have hindered in vivo research on the kinetics of primate behaviors on vertical substrates. We describe a simple, inexpensive apparatus for recording forces from primate behaviors on vertical substrates. The apparatus includes an instrumented beam fastened directly to a horizontal force platform and a surrounding vertical substrate that does not contact the instrumented beam or platform. The contact piece at the end of the instrumented beam is positioned flush with the noninstrumented vertical substrate, and reaction forces elicited on this instrumented section are directed to the force platform. Because most of the vertical substrate is not instrumented, we can isolate and record forces from a single limb or jaw during a behavior. Biewener and Full ([1992] Biomechanics Structures and Positions: A Practical Approach; New York: Oxford University press, p. 45-73) gave seven criteria to consider when designing a customized force-recording device. Where appropriate, we tested if our apparatus met their criteria. The apparatus accurately records forces in three orthogonal directions, has low cross-talk, maintains a high frequency response, exhibits a linear response up to at least 200 Newtons, and displays a uniform response to a given force across the instrumented contact piece. Our design does not easily facilitate the identification of the point of force application. Therefore, joint moments cannot be easily calculated. This limitation, however, does not affect the apparatus's ability to accurately record the magnitude and direction of a force (as shown by other tests). We developed this apparatus to measure jaw forces during tree gouging in common marmosets (Callithrix jacchus), but the general design can be readily modified to study a variety of primate behaviors on vertical substrates.     

The use of on‐animal acoustical recording devices for studying animal behavior

Audio recordings made from free‐ranging animals can be used to investigate aspects of physiology, behavior, and ecology through acoustic signal processing. On‐animal acoustical monitoring applications allow continuous remote data collection, and can serve to address questions across temporal and spatial scales. We report on the design of an inexpensive collar‐mounted recording device and present data on the activity budget of wild mule deer (Odocoileus hemionus) derived from these devices applied for a 2‐week period. Over 3300 h of acoustical recordings were collected from 10 deer on their winter range in a natural gas extraction field in northwestern Colorado. Analysis of a subset of the data indicated deer spent approximately 33.5% of their time browsing, 20.8% of their time processing food through mastication, and nearly 38.3% of their time digesting through rumination, with marked differences in diel patterning of these activities. Systematic auditory vigilance was a salient activity when masticating, and these data offer options for quantifying wildlife responses to varying listening conditions and predation risk. These results (validated using direct observation) demonstrate that acoustical monitoring is a viable and accurate method for characterizing individual time budgets and behaviors of ungulates, and may provide new insight into the ways external forces affect wildlife behavior.     

Single-channel recording of ligand-gated ion channels

Electrophysiological recording of single-channel currents is the most direct method available for obtaining detailed and precise information about the kinetic behavior of ion channels. A wide variety of cell types can be used for single-channel recording, but to obtain the highest resolution of the briefest channel opening and closing events, low-noise recordings, coupled with a minimal filtering frequency, are required. Here, we present a protocol designed to help those with some electrophysiological expertise who wish to explore the properties of native and recombinant single ligand-gated ion channels. We have focused on the practical aspects of recording single GABA channels from cell-attached and outside-out patches and also introduced some of the preliminary considerations that are necessary for the analysis of single-channel data, including an introduction to single-channel analysis software.     

The functional role of octopaminergic neurons in insect motor behavior

The role of efferent, octopaminergic dorsal unpaired median (DUM) neurons in insects is examined by recording from them during motor behaviour. This population of neuromodulatory neurons is divided into sub-populations which are specifically activated or inhibited during ongoing motor behavior. These neurons are always activated in parallel to the respective motor circuits, and in addition to their modulatory effects on synaptic transmission may also cause metabolic changes in their target tissues.     

The distribution of solid liposomes of different size on the surface of epithelial cells

Interaction of large multilamellar and small sonicated liposomes with epithelial sheets in culture has been studied. Liposomes were comprised of dipalmitoyl- or distearoylphosphatidylcholine and were solid at 37 degrees. Sodium fluorescein was entrapped in liposomes and their localization on the cell surface was studied by fluorescence microscopy. The distribution of liposomes depends on their size. Large liposomes adhere uniformly all over the surface of the sheet, which proved to be nonadhesive either for other cells or for inert particles. Small liposomes adhere mostly along the margins of the sheet. When the cells are partly detached from each other by mild EDTA (ethylene diamine tetraacetic acid) treatment, the newly accessible cell margins become capable of binding small liposomes. The binding of both types of liposomes is trypsin-sensitive; however large liposomes, bound along the edges of the cellular sheet, are removed slower than those adhering to the inner parts of the sheet. Competition experiments show that large and small liposomes bind to the same sites on the surface of the sheet. Liposome-acceptor proteins are suggested to exist on the surface of epithelial cells. We suppose that these proteins are concentrated along the cell margins and are involved in cell-cell contacts.     

Digital registration and analysis of visual information in behavioral experiment

A new software for recording, registration and computer analysis of behavior of laboratory animals, EthoStudio, has been developed. The software allows the latency, number and duration of up to 10 behavioral patterns to be simultaneously registered with parallel recording of video on a hard disk. The video records can be played back to track animal locomotion using a computer. EthoStudio was applied to study mouse behavior in the open field test. The numbers of rearings and groomings as well as accumulated time of grooming were interactively assayed by pressing and holding buttons on a computer keyboard. The number of squares crossed, the number of entries into the center and the time spend therein were assayed by computer analysis. EthoStudio can be applied to objectively study various kinds of animal behavior with reliability and accuracy necessary for modern ethological, physiological and pharmacological experiment.     

Multidimensional recording (MDR) and data sharing: an ecological open research and educational platform for neuroscience

Primate neurophysiology has revealed various neural mechanisms at the single-cell level and population level. However, because recording techniques have not been updated for several decades, the types of experimental design that can be applied in the emerging field of social neuroscience are limited, in particular those involving interactions within a realistic social environment. To address these limitations and allow more freedom in experimental design to understand dynamic adaptive neural functions, multidimensional recording (MDR) was developed. MDR obtains behavioral, neural, eye position, and other biological data simultaneously by using integrated multiple recording systems. MDR gives a wide degree of freedom in experimental design because the level of behavioral restraint is adjustable depending on the experimental requirements while still maintaining the signal quality. The biggest advantage of MDR is that it can provide a stable neural signal at higher temporal resolution at the network level from multiple subjects for months, which no other method can provide. Conventional event-related analysis of MDR data shows results consistent with previous findings, whereas new methods of analysis can reveal network mechanisms that could not have been investigated previously. MDR data are now shared in the public server Neurotycho.org. These recording and sharing methods support an ecological system that is open to everyone and will be a valuable and powerful research/educational platform for understanding the dynamic mechanisms of neural networks.     

Telemetered heart rate responses of the rat during free and learned behavior

To assess the effect of recording on the heart rate and on the behavior of the free moving rat, the electrocardiogram was recorded with the aid of radiotelemetry or through wire leads attached to the animal in different behavioral situations. Adaptation of heart rate both within and across sessions in the home cage and in a novel environment (open-field) was observed in rats bearing a transmitter. No adaptation occurred in rats attached to wire leads. The pattern of heart rate changes appeared to be independent of the mode of recording during passive avoidance behavior and when the rats displayed various motor activities like walking, sniffing, rearing or grooming in the open-field. The heart rate of rats attached to wire leads, however, was always higher. These data indicate that radiotelemetry is preferable to the use of wire connections for recording cardiac activity during behavioral studies in the rat.     

Effects of androgens and estrogens on crowings and distress callings in male Japanese quail, Coturnix japonica

In male Japanese quail, crowing behavior is considered to be strictly androgen-dependent. It was previously shown that in chicks, treatment with either testosterone or 5alpha-dihydrotestosterone (5alpha-DHT; a non-aromatizable androgen) induced crowing with motivation for distress calling in acutely isolated conditions. Many studies, however, have shown that the potencies of testosterone and 5alpha-DHT in activating crowing in castrated males are different. To clarify the effects of androgenic and estrogenic actions on the production of crows and distress calls, we injected quail daily from 11 to 42 days after hatching (Day 11 to 42) with testosterone propionate (TP), 5alpha-DHT, estradiol benzoate (EB) or vehicle and examined their calling behaviors both in a recording chamber (acutely isolated conditions) and in their home-cages (well-acclimated conditions). Both TP- and 5alpha-DHT-treated birds began to crow by Day 13 when isolated in the recording chamber. The TP-treated birds, however, crowed less frequently than 5alpha-DHT-treated ones. This, combined with the observations that distress calling was strongly inhibited in EB-treated birds, suggests that estrogen converted from testosterone may inhibit the motivation for distress calling. On the other hand, after chronic treatment of TP, but not of 5alpha-DHT, birds began to crow intensely in their home-cages earlier than vehicle treated controls, suggesting that estrogen is needed to initiate crowing behavior in sexually active males. Taken together, it is suggested that estrogenic actions affect the motivation underlying vocal behaviors, while the androgenic action is indispensable in generating crowing.