Petrolatum

Extracellular recordings of the discharges of a prominent interneuron, the AN1-neuron, were used to examine both the tuning of the hearing system and the peripheral directionality. Action potential activity was recorded with extracellular tungsten hook-electrodes placed at the connectives between prothoracic and suboesophageal ganglia. Insects were shortly anesthesized with chlorethylene and fixed ventral side up on a thin (1 mm) platform with dental wax. The forelegs were fixed in the natural walking position onto thin wires (diameter 0.6 mm). The preparation was sealed with petroleum jelly to prevent desiccation of the connectives. AN1-discharges were amplified by a custom-made amplifier, visualized on an oscilloscope (Agilent 54616B) and monitored through headphones. The threshold for each stimulus was defined as the SPL which elicited at least one AP in each syllable in at least three out of five stimulations.
Neurophysiological experiments were performed in an acoustically isolated Faraday-cage at room temperature between 21–23°C. A speaker (Raveland MHX 138) was placed at a distance of 40 cm, at an angular separation of 30° left or 30° right of the longitudinal body axis. Model songs identical to those used in behaviour were generated, amplified and attenuated in steps of 1 dB using a Tucker-Davis system (Alachua, Florida). The tuning of AN1 was determined with ipsilateral stimulation at frequencies ranging from 3.5 to 6 kHz, in increments of 100 or 500 Hz. In order to measure only the directionality provided by the anatomical arrangement of the acoustic tracheae in the periphery, inhibitory central nervous interactions were eliminated by cutting the contralateral leg nerve with the afferent auditory fibres of the opposite ear. IIDs were calculated by measuring the thresholds of the AN1-neuron for frequencies between 3.5 to 6 kHz with ipsilateral and contralateral stimulation at an angular frontal deviation of 30°. The threshold differences between ipsi- and contralateral stimulation represent the IID for a given frequency at a stimulation angle of ±30°, similar to biophysical measurements using laser-vibrometry [27] ). Pronotum width was measured as an index of structural body size to the nearest 0.1 mm, using a digital calliper.

Semi-field experiments were started at 20.00 when it was completely dark. This helped to exclude the possibility of experimental mosquitoes released from the centre of the screen-house from responding to a unidirectional source of light occasioned by sunset. The start of experiments in darkness was not necessary under field conditions (i.e.18:30–06:30) because the wild mosquitoes lured to trapping devices originated randomly from sources located in different directions.
All MM-X traps were operated on 12 V. Vaseline pure petroleum jelly was also applied on suspension wire bars and electrical cables to prevent ants from preying on mosquitoes caught in the MM-X traps. Baited traps and an unbaited MM-X trap were randomly assigned and alternated daily between trap positions to eliminate confounding effects associated with site. A data logger (Tinytag® Ultra, model TGU-1500, INTAB Benelux, the Netherlands) was used to record ambient temperature and RH at an interval of 30 min. To terminate individual experiments, a plug was inserted into the outer tube of the MM-X trap, the CO₂ supply cut off, and power switched off (semi-field) or traps disconnected from batteries (field study). Latex gloves were worn during preparation of refined sugar/molasses-yeast mixtures, nylon strips, application of attractants on nylon strips and baiting of traps to avoid contamination with human volatiles or other odorant compounds. Traps containing mosquitoes were placed in a refrigerator at -4°C for 30 min. Immobilized mosquitoes were collected from each trap, counted and recorded. Thereafter, traps were cleaned using 70% methanol (to remove residual odours) between experiments. A manual, hand held aspirator was used to collect untrapped, free-flying mosquitoes from the screen-walled greenhouse and killed. The sand-filled floor of the greenhouse was moistened daily to enhance survival of mosquitoes.

All experiments were performed in accordance with relevant guidelines and regulations. Eight weeks old female BALB/c mice were used for this study (Harlan Laboratories, Indianapolis, IN). All animal procedures were approved by Purdue University Animal Care and Use Committee (PACUC). The murine model of MRSA skin infection has been described before52 . Mice were injected intradermally with 40 μl of MRSA USA300 (1.65 × 10⁸) CFU per mouse. Forty-eight hours after infection and formation of open wound, the mice were divided into eight groups (n = 5). Four groups were treated topically with either 0.5%, 1%, or 2% ebselen in petroleum jelly (ointment- skin protectant) or 1% ebselen in lipoderm (dermal and transdermal delivery cream base). Two groups received the vehicles alone (petroleum jelly or lipoderm). One group was treated topically with 2% mupirocin in petroleum jelly and the last group was treated orally with linezolid (25 mg/kg). All groups were treated twice a day for 5 days. Twenty-four hours after the last treatment, the area around the wound was lightly swabbed with 70% ethanol and the wound was excised for bacterial counting on MSA after homogenization.

A tongue-shaped flap was created on the radial wall of the 5th digit, with the longitudinal edges not exceeding the radial surface of the digit, and the distal edge made slightly beyond the PIP joint line. To ensure full coverage of the volar skin defect, the flap was made 2 mm larger in diameter than the recipient site (Fig. 2).Figure 2

Representative illustrations of camptodactyly of the 5th digit. (a) Frontal and lateral views of the 5th digit before surgery. (b) Design of the tongue-shaped flap, with the longitudinal edges limited within the radial surface, and the distal edge made slightly exceeding the proximal interphalangeal joint line. (c) The volar incision. (d) The lateral view of the digit flap transfer, with direct suturing performed for closure of the donor site. (e) The volar view of the digit after flap transfer, with complete coverage of the volar skin defect.

While creating the edges of the flap, care was taken to preserve the perforating blood vessels of the proper palmar digital arteries, as well as the proper palmar digital nerves.
Sequential release of affected soft tissues was performed in the following order—skin, subcutaneous fibrous fascia, flexor digitorum superficialis tendon, lumbrical muscle insertions if present, and volar plate. The degree of passive extension of the PIP joint was repeatedly tested, and surgical release was considered complete upon achieving full passive extension of the joint. Kirschner (K)-wire fixation was performed following volar plate release.
The radial flap was rotated 90° to cover the volar skin defect, and direct suturing was performed to close the donor site. Free skin grafting was indicated in the presence of high suture tension.
Mupirocin ointment and petroleum jelly (Vaseline) were subsequently applied, and the wound was wrapped with clean dressing. All digits were immobilized in the extended position with a cast for three weeks.

Static compression tests (strain rate at 10⁻³ s⁻¹ and 10⁻² s⁻¹) were conducted on an electronic material testing machine (Instron^® Model 5,969; Instron, United States) with a load cell capacity of 10 kN. Loading rates were calculated according to the longitudinal length of each specimen and strain rate. All tests were performed at room temperature with occasional sprinkling of saline to keep the samples hydrated. The contact surface between the sample and the platen was lubricated with petroleum jelly, and it was confirmed that the two loading surfaces of the sample were parallel and no horizontal displacement occurred during the compression process.
Dynamic compression tests of bone sample were performed by a modified Split Hopkinson Bar (SHPB). The striker bar, incident bar, and transmitter bar have the same diameter of 14.5 mm and different lengths of 200 mm, 1,500 mm, and 1,500 mm (Aluminum 7A04 bars). The bone sample was sandwiched between the incident bar and the transmitter bar. Both semiconductor strain gauges (120 Ω, GmbH, Germany) across the bar diameter at a bar location were connected in series to the same leg in the Wheatstone bridge to offset the influence of bending stress. The strain gauges were connected to a signal conditioner amplifier and data was recorded with a Dynamic Signal Acquisition and Analysis System (Donghua, China) at a sampling rate of 1 MHz. In order to achieve stress equilibrium and constant strain rate conditions, a pulse shaper processed from paper jam into a uniform cylinder was placed between the striker bar and the incident bar. According to theory of the one-dimensional wave transmission, the strain rate increases with the diameter of the pulse shaper. Similarly, the slope of the loading wave increases with the striker speed and loading time. Thus, a uniformly machined copper cylinder pulse shaper was used at different sizes for filtering high frequency waves. A schematic diagram of the experimental procedure is shown in Figure 1. The representative curves obtained are shown in Figure 2A. Over the entire pulse duration, the signal is consistent with Eq. (1). Therefore, it can be considered that the stress equilibrium is maintained in the experiment. ${\epsilon }_I+{\epsilon }_R={\epsilon }_T$ where ${\epsilon }_I$ is the incident wave, ${\epsilon }_R$ is the reflected wave and ${\epsilon }_T$ is the transmitted wave. The stress-strain curve calculated from the pulse signal is shown in Figure 2B. The strain rate rises rapidly to the expected constant strain rate, and then remains constant until specimen failure. The strain occurring in the specimen at a constant strain rate condition exceeds 70% of the total strain. Therefore, the method designed in this paper is effective in reflecting the compressive mechanical properties of cortical bone at the preset strain rate.

The ABL is designed following Bartlett et al. [42 (link)], and consists of a soft silicone matrix that is internally reinforced with a polyester mesh. To produce the ABL, we clamped rectangular midge mesh (30 fibres in tail direction and 18 fibres in width direction per square inch) between two 0.5 mm thick polystyrene plates with 5 cm × 5 cm large rectangular cut-outs, which were placed on a smooth glass plate. Gaps between glass, mesh and polystyrene were sealed with petroleum jelly. We prepared silicone rubber (Resion rubber SR1, shore hardness 8A, polyestershoppen.nl) at a part A to B weight ratio of 1 : 5, coloured the silicone with white pigment (1 wt% white silicone pigment, siliconesandmore.nl) for later contact area imaging, and filled the cut-outs with silicone. Air pockets were removed manually if present. After curing, the mesh-reinforced silicone was removed from the polystyrene plates and glass, cut into 5 cm × 5 cm large adhesives with free mesh tails, and cleaned with isopropanol and water. The silicone surface that cured onto the glass served as contact surface in the later experiments.
To assess variability of the manufactured ABL specimen, we measured their friction performance on a glass cylinder, whose long axis runs parallel to the ABL’s tail (see §2.2 for setup details). When reusing a single specimen for 30 consecutive repetitions, static and dynamic friction were $46.0\pm 1.3\hspace{0.17em}\mathrm{N}$ and $33.3\pm 1.6\hspace{0.17em}\mathrm{N}$ , respectively (mean ± s.d.), corresponding to an intra-specimen variability below 4.3%. Additionally, we measured all 26 ABL specimens used in the later experiments to test for inter-specimen variability. With a static and dynamic friction of $53.3\pm 5.5\hspace{0.17em}\mathrm{N}$ and $33.4\pm 1.1\hspace{0.17em}\mathrm{N}$ , variability between specimens was below 10.4%.