How do gastropods grow synchronized shell sculpture? Effect of experimental varix manipulations on shell growth by Ceratostoma foliatum (Muricidae: Ocenebrinae)

Gastropod shells display striking patterns in both color and sculpture, but rather little is known about the developmental mechanisms that produce those patterns. Here, we tested a physical feedback hypothesis for how snails control spatial patterning of shell sculpture. Varices—a form of synchronized, blade‐like axial sculpture—are produced at regular intervals around the shell and often aligned closely between adjacent whorls. Older varices were believed to provide a spatial cue about where to position a new varix. To test this hypothesis, we manipulated physical cues by cutting off varices or attaching new ones to the body whorl of individuals of Ceratostoma foliatum, and then allowing snails to grow a new varix. We found that previous varices on the shell were neither necessary nor sufficient to induce a new varix at a particular location. However, the position of older varices did appear to affect the fine tuning of subsequent varix placement. The results of our experiments therefore suggest that varix synchrony arises mainly from some internal mechanism that yields a standardized amount of spiral growth per growth spurt. We also found that shell damage can induce varix production in unusual or aberrant locations during subsequent shell growth.     

In ovo electroporations of HH stage 10 chicken embryos

Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology. With the advent of molecular biological techniques, the chick has become a useful system in which to study gene regulation and function. By electroporating DNA or RNA constructs into the developing chicken embryo, genes can be expressed or knocked down in order to analyze in vivo gene function. Similarly, reporter constructs can be used for fate mapping or to examine putative gene regulatory elements. Compared to similar experiments in mouse, chick electroporation has the advantages of being quick, easy and inexpensive. This video demonstrates first how to make a window in the eggshell to manipulate the embryo. Next, the embryo is visualized with a dilute solution of India ink injected below the embryo. A glass needle and pipette are used to inject DNA and Fast Green dye into the developing neural tube, then platinum electrodes are placed parallel to the embryo and short electrical pulses are administered with a pulse generator. Finally, the egg is sealed with tape and placed back into an incubator for further development. Additionally, the video shows proper egg storage and handling and discusses possible causes of embryo loss following electroporation.     

Elastic plastic tube for detecting exfoliative cancer cells in the esophagus.

A simple device for sampling esophageal lesions is described. The instrument, which is easy and inexpensive to make, consists of a single-lumen plastic tube covered with cotton thread. Once swallowed, the end of the tube can be drawn into a loop. The device has been used at locations with a high incidence of esophageal cancer and at our tumor hospital. A series of 746 cases examined had no complications. The tube has a positive diagnostic rate comparable to that of roentgenography, 96%, and seemed acceptable to both physicians and patients. When an obstruction is present, the instrument can easily pass through it and obtain exfoliated cancer cells, while the conventional method, the balloon cytologic smear, is unable to do so.     

Varix of the digit

Varix of the digit is a rarely described lesion. Over 3 years, eight patients were diagnosed as having this abnormality. Surgical confirmation of the identity of the lesion was accomplished in six of these patients. Repetitive trauma, mechanical compression, and aging phlebectasia have all been associated with the pathogenesis of a digital varix. Thrombophlebitic and thrombotic changes occurring within the varix may lead to an erroneous assessment and clinical confusion. In such instances, a tourniquet test has been useful in demonstrating the presence of a varix. Excision of a symptomatic lesion has proven to be curative for all six patients. Two patients have been observed for 3 years with little change and no resolution of the varix. It is probable that varix of the digit is a more common lesion than indicated by a review of the literature.     

Varix of the vortex ampulla: a dynamic phenomenon

A vortex varix consists of a bulging of the vortex ampulla and overlying retina. This dynamic retinal finding can change with the patient's direction of gaze. The clinician can often visualize a red, elevated lesion that appears and disappears thus, making the diagnosis confusing. An overview of vortex varices, methods of diagnosis and differential diagnoses will be discussed.     

A simple and disposable sweat collector.

Apart from in cystic fibrosis, where sweat analysis provides valuable diagnostic information, sweat yields remain an overlooked biological fluid. Technical problems (dilution, condensation, contamination, evaporation, etc.) linked to currently available collection procedures are of concern and thwart their use. To overcome some of these technical difficulties, an original sweat-collection technique is described. A collection capsule is created inside a flexible, adhesive and disposable anchoring membrane pasted onto the skin. A fluid-tight window is positioned in the upper part of the pocket and gives access to its content. Through the collection window, complete emptying of the sweat collector can be achieved repeatedly by suction using a vacutainer tube inserted in a tube holder equipped with a long dull needle. With prior addition of a suitable marker, fractional samplings can also be performed using a precision micropipette. This collecting method allows for kinetic studies on sweat rate and sweat content. The limited bias-inducing manipulations linked to the described technique, coupled with the ease of performing kinetic studies on sweat volume and content, make this original tool a reliable and accurate sweat-collection technique.     

Experimental Study of a Starting Vortex Ring Generated by a Thin Circular Disk

The formation process of vortex ring generated by a thin circular disk was studied experimentally in this paper. A thin circular disk installed a linear motion stage was used to generate the vortex rings. Digital Particle Image Velocimetry (DPIV) was used to measure the velocity and vorticity fields. The finite-time Lyapunov exponent field corresponding to the vortex flow was computed to identify Lagrangian coherent structures of the starting vortex. The results reveal the existence of a flux window between repelling Lagrangian Coherent Structures (rLCS) and attracting Lagrangian Coherent Structures (aLCS), through which the shear flow is entrained into the vortex. The flux window is closed gradually during the starting process. Once the flux window shut down, the formation process of the vortex ring finishes, as the shear flow with vorticity cannot be entrained in the vortex ring.     

Visualizing the Structural Changes of Bacteriophage Epsilon15 and Its Salmonella Host during Infection

The efficient mechanism by which double-stranded DNA bacteriophages deliver their chromosome across the outer membrane, cell wall, and inner membrane of Gram-negative bacteria remains obscure. Advances in single-particle electron cryomicroscopy have recently revealed details of the organization of the DNA injection apparatus within the mature virion for various bacteriophages, including epsilon15 (?15) and P-SSP7. We have used electron cryotomography and three-dimensional subvolume averaging to capture snapshots of ?15 infecting its host Salmonella anatum. These structures suggest the following stages of infection. In the first stage, the tailspikes of ?15 attach to the surface of the host cell. Next, ?15's tail hub attaches to a putative cell receptor and establishes a tunnel through which the injection core proteins behind the portal exit the virion. A tube spanning the periplasmic space is formed for viral DNA passage, presumably from the rearrangement of core proteins or from cellular components. This tube would direct the DNA into the cytoplasm and protect it from periplasmic nucleases. Once the DNA has been injected into the cell, the tube and portal seals, and the empty bacteriophage remains at the cell surface.     

Percutaneous nephrolithotomy.

A new method of removing calculi from the renal collecting system, in which the large and traumatic incision in the loin currently used is unnecessary, was attempted in 31 patients. Small dilators were introduced over a guide wire through a nephrostomy tube into the renal pelvis and a catheter inserted. The track was dilated in stages and two days later the nephrostomy tube was removed and a cystoscope introduced into the interior of the kidney. A stone basket was introduced down the operating channel of the cystoscope and maneuvered to secure the stone; the cystoscope, stone basket, and stone were then removed. The procedure failed in 11 of the 31 patients, in five because the needle could not be placed accurately initially and in six because the stone could not be removed despite the establishment of a nephrostomy track. In the remaining 20 patients the procedure was successful. This procedure is far less traumatic than the conventional operation. With the development of a flexible nephroscope and an ultrasonic stone disintegrator it will be possible also to remove larger stones and stones in the peripheral calices using this method.     

Anticipatory Smooth Eye Movements in Autism Spectrum Disorder

Smooth pursuit eye movements are important for vision because they maintain the line of sight on targets that move smoothly within the visual field. Smooth pursuit is driven by neural representations of motion, including a surprisingly strong influence of high-level signals representing expected motion. We studied anticipatory smooth eye movements (defined as smooth eye movements in the direction of expected future motion) produced by salient visual cues in a group of high-functioning observers with Autism Spectrum Disorder (ASD), a condition that has been associated with difficulties in either generating predictions, or translating predictions into effective motor commands. Eye movements were recorded while participants pursued the motion of a disc that moved within an outline drawing of an inverted Y-shaped tube. The cue to the motion path was a visual barrier that blocked the untraveled branch (right or left) of the tube. ASD participants showed strong anticipatory smooth eye movements whose velocity was the same as that of a group of neurotypical participants. Anticipatory smooth eye movements appeared on the very first cued trial, indicating that trial-by-trial learning was not responsible for the responses. These results are significant because they show that anticipatory capacities are intact in high-functioning ASD in cases where the cue to the motion path is highly salient and unambiguous. Once the ability to generate anticipatory pursuit is demonstrated, the study of the anticipatory responses with a variety of types of cues provides a window into the perceptual or cognitive processes that underlie the interpretation of events in natural environments or social situations.     

Wave dissipation in flexible tubes in the time domain: in vitro model of arterial waves

Earlier work of wave dissipation in flexible tubes and arteries has been carried out predominantly in the frequency domain and most of the studies used the measured pressure waveform for presenting the results. In this work we investigate the pattern of wave dissipation in the time domain using the separated forward and backward travelling waves in flexible tubes. We tested four sizes of latex tubes of 2m in length each, where a single semi-sinusoidal in shape, pressure wave, was produced at the inlet of each tube. Simultaneous measurements of pressure and flow waveforms were recorded every 5cm along the tubes and wave speed was determined using the pressure-velocity loop method (PU-loop). The measured data and wave speed were used to separate the pressure waveform and wave intensity, into their forward and backward directions, using wave intensity analysis (WIA). Also, the energy carried by the wave was calculated by integrating the relevant area under the wave intensity curve. The peak of the measured pressure waveform increased downstream, however, the peak of the separated forward pressure waveform decreased exponentially along the tube. Wave intensity and energy also dissipated exponentially along the travelling distance. The peaks of the separated pressure and wave intensity decreased in the forward in a similar exponential way to that in the backward direction in all four tube sizes. Also, the smaller the size of the tube the greater wave dissipation it caused. We conclude that wave separation is useful in studying wave dissipation in elastic tubes, and WIA provides a convenient method for determining the dissipation of the energy carried by the wave along the travelled distance. The separated pressure waveform, wave intensity and wave energy dissipate exponentially with the travelling distance, and wave dissipation varies conversely with the diameter of elastic tubes.     

BrdU assay for neurogenesis in rodents

Neurogenesis within the adult central nervous system is demonstrated using an exogenous cell tracer, 5'-bromo-2'-deoxyuridine (BrdU), in combination with endogenous neuronal markers. Specific primary antibodies raised against these markers are widely available and their visualization is possible with the use of fluorescently tagged secondary antibodies. BrdU is a thymidine analog that incorporates into dividing cells during DNA synthesis. Once incorporated into the new DNA, BrdU will remain in place and be passed down to daughter cells following division. Typically, BrdU is injected intraperitoneally. Different survival times required by the desired experimental time-line will yield data on specific phases of neurogenesis: proliferation, differentiation and maturation. One of the drawbacks of using BrdU is the dependence on a stressful injection procedure and uncertain penetration of the targeted cells with a uniform concentration of the compound. Thus, for experiments requiring measurements of cell proliferation, Ki67 can be used as an acceptable alternative. The protocol takes 3-5 d, allowing for sectioning and staining.     

An autoradiographic study of the distribution of fibers from the dorsal motor nucleus of the vagus to the digestive tube of the rat

At the present time, complete agreement on the origin and course of parasympathetic preganglionic fibers to the alimentary canal has not been reached. The purpose of this study was to trace vagal fibers to the abdominal cavity and to follow the distribution of these fibers to the digestive tube. The technique used was to label neurons in the dorsal motor nucleus of the vagus (DMX) with 3H-leucine and then to follow the orthograde transport. 16 albino rats were used in this experiment. The right DMX in one group of rats and the left DMX in the other group was injected with 25 microCi of 3H-leucine in three injections. The injection sites and tissue sections from various areas of the digestive tube were processed for autoradiography. A heavy label was observed in the injection site and it could be traced down the vagus nerve through the thorax into the abdomen. Labelled vagal fibers were found in the parasympathetic ganglia of the stomach, small intestine and colon.     

Nuclear magnetic resonance-based quantification of organic diphosphates

Phosphorylated compounds are ubiquitous in life. Given their central role, many such substrates and analogs have been prepared for subsequent evaluation. Prior to biological experiments, it is typically necessary to determine the concentration of the target molecule in solution. Here we describe a method where concentrations of stock solutions of organic diphosphates and bisphosphonates are quantified using ³¹P nuclear magnetic resonance (NMR) spectroscopy with standard instrumentation using a capillary tube with a secondary standard. The method is specific and is applicable down to a concentration of 200 μM. The capillary tube provides the reference peak for quantification and deuterated solvent for locking.     

Multi-scale parametric spectral analysis for exon detection in DNA sequences based on forward-backward linear prediction and singular value decomposition of the double-base curves

This paper presents a new method for exon detection in DNA sequences based on multi-scale parametric spectral analysis. A forward-backward linear prediction (FBLP) with the singular value decomposition (SVD) algorithm FBLP-SVD is applied to the double-base curves (DB-curves) of a DNA sequence using a variable moving window sizes to estimate the signal spectrum at multiple scales. Simulations are done on short human genes in the range of 11bp to 2032bp and the results show that our proposed method out-performs the classical Fourier transform method. The multi-scale approach is shown to be more effective than using a single scale with a fixed window size. In addition, our method is flexible as it requires no training data.     

Modeling of Surface-Tension-Driven Flow of Blood in Capillary Tubes

Surface-tension-driven blood flow into a capillary tube, as in some medical devices, is studied. In a previous article, we considered the early stages of the entry flow from a drop of blood into a capillary, and solved the problem analytically under the assumption that the resistance of the air is negligible. In the present note we consider a capillary tube of finite length, with the far end containing a small window which opens to the atmosphere. The dynamic reverberation of the air in the capillary tube is analyzed in conjunction with the dynamics of the blood. Existing computing programs are used to solve the Navier-Stokes equations. The interface is characterized by the surface tension between the blood and the air, and the contact angle at the triple point where the air-blood interface meets the capillary tube wall. The results tell us how good our earlier simplified analysis is. The new numerical results show that the smaller the window, the larger is the effect of aerodynamic reverberation. However, even for a window as small as 4% of the capillary cross section, and located at the end of the capillary, the difference of the time of arrival of the interface at the window is less than 5%.     

胚泡着床窗口的分子调控

着床窗口是指当胚胎发育到胚泡阶段时,子宫也增殖和分化到可接受状态,二者相互作用使胚泡着床的短暂时间.雌激素和孕酮是该过程的综合调控分子,它们通过多种局部信号分子的介导,使子宫中的各种细胞类型增殖、分化,为着床窗口的开放做出相互协调的反应.子宫与胚胎在着床窗口通过前列腺素、组织胺、降钙素、多种细胞因子和生长因子的旁分泌作用进行分子对话,使胚泡滋养层与子宫内膜上皮发生附着反应.着床窗口一旦开放,即自动向非接受态转化.     

Acute respiratory insufficiency after endoscopy for bleeding oesophageal varices.

Two patients with alcoholic liver disease and gross ascites underwent endoscopic injection and compression by Sengstaken tube of oesophageal varices under general anaesthesia. Postoperatively both patients developed acute respiratory failure, which resolved after air had been aspirated from the stomach via the Sengstaken tube. All air should be aspirated at the end of the procedure in patients with ascites who undergo endoscopy, and respiration should be carefully supervised postoperatively.     

An automatic method for the measurement of the effect of short stresses on the microbial ATP pool

An automatic method for measuring the effects of brief contacts between microorganisms and toxic agents is described.In this method, both a microbial culture and a toxic solution are injected into a mixing tube, until inhibition of bacteria and ATP extraction take place.The range of the contact times, defined as the residence periods of the cultures in the mixing tube, is quite wide (from 0.3 to 16 s), with increased coefficient of variation (from 5 to 14%) for the lower times.     

Determination of monensin, salinomycin and narasin in muscle, liver and eggs from domestic fowl using liquid chromatography-electrospray mass spectrometry

A method is presented for the simultaneous determination of the three ionophores, monensin, salinomycin and narasin in muscle, liver and eggs from domestic fowl. Samples are homogenised in methanol and any ionophores present are extracted into toluene-hexane. The extracts are concentrated and injected into a bench-top electrospray LC-MS system. Chromatography is carried out using an end-capped reversed-phase column, with a mobile phase consisting a mixture of acetonitrile, methanol, tetrahydrofuran, trifluoroacetic acid and water. Using single-ion monitoring, the ionophores can be detected down to the 1 ng/g level.