It is well established that parasites in the phylum Nematomorpha induce suicide behavior of their insect hosts to bring adult worms to the appropriate habitat for emergence. It is not well established, however, whether other nematomorph-induced behavioral alterations occur before worm emergence. The purpose of our study was to evaluate the effect of the nematomorph Paragordius varius on the calling behavior of the male house cricket Acheta domesticus. We hypothesized that cricket calling, an energetically expensive and risky behavior, would be a potential target for nematomorph-induced behavioral alterations. We assessed if and how infection with P. varius affects A. domesticus calling behavior and whether the presence of wings at time of exposure to P. varius influenced changes in calling behavior. We recorded the calling behavior of male A. domesticus over the course of their infection after exposure to P. various before or after wing development. Additionally, we assessed whether winged crickets were "callers'' or "noncallers'' before exposure. We found that regardless of cricket developmental stage (or age) at time of infection, infected crickets spent significantly less time calling than their uninfected counterparts but only during the later stages of infection. Developmental stage at infection did affect whether crickets became callers: when infected before wing development significantly more uninfected crickets initiated calling; there was no difference between infected and uninfected crickets when infected as winged adults. Infection was a factor in whether callers stopped calling, with more infected crickets ceasing to call than uninfected crickets. This is the first study to show that infection with nematomorphs affects calling behavior of their insect host. Cricket calling behavior is immensely complex and although it was difficult to elucidate the adaptive nature of these parasite-induced behavioral changes, this study lays the groundwork for future studies to begin teasing out the factors that will help make the determination between side effect of infection or parasite/host adaptation.

BACKGROUND:Access to timely and accurate diagnostic tests has a significant impact in the management of diseases of global concern such as malaria. While molecular diagnostics satisfy this need effectively in developed countries, barriers in technology, reagent storage, cost and expertise have hampered the introduction of these methods in developing countries. In this study a simple, lab-on-chip PCR diagnostic was created for malaria that overcomes these challenges.METHODS:The platform consists of a disposable plastic chip and a low-cost, portable, real-time PCR machine. The chip contains a desiccated hydrogel with reagents needed for Plasmodium specific PCR. Chips can be stored at room temperature and used on demand by rehydrating the gel with unprocessed blood, avoiding the need for sample preparation. These chips were run on a custom-built instrument containing a Peltier element for thermal cycling and a laser/camera setup for amplicon detection.RESULTS:This diagnostic was capable of detecting all Plasmodium species with a limit of detection for Plasmodium falciparum of 2 parasites/μL of blood. This exceeds the sensitivity of microscopy, the current standard for diagnosis in the field, by ten to fifty-fold. In a blind panel of 188 patient samples from a hyper-endemic region of malaria transmission in Uganda, the diagnostic had high sensitivity (97.4%) and specificity (93.8%) versus conventional real-time PCR. The test also distinguished the two most prevalent malaria species in mixed infections, P. falciparum and Plasmodium vivax. A second blind panel of 38 patient samples was tested on a streamlined instrument with LED-based excitation, achieving a sensitivity of 96.7% and a specificity of 100%.CONCLUSIONS:These results describe the development of a lab-on-chip PCR diagnostic from initial concept to ready-for-manufacture design. This platform will be useful in front-line malaria diagnosis, elimination programmes, and clinical trials. Furthermore, test chips can be adapted to detect other pathogens for a differential diagnosis in the field. The flexibility, reliability, and robustness of this technology hold much promise for its use as a novel molecular diagnostic platform in developing countries.

This work describes a self-contained, simple, disposable, and inexpensive gel capillary cassette for DNA amplification in near point of care settings. The cassette avoids the need for pumps or valves during raw sample delivery or polymerase chain reaction (PCR) amplification steps. The cassette contains capillary reaction units that can be stored at room temperature for up to 3 months. The current cassette configuration format simultaneously tests up to 16 patients for two or more targets, accommodates different sample types on the same cassette, has integrated positive and negative controls and allows flexibility for multiple geometries. PCR reagents in the cassette are desiccated to allow storage at room temperature with rehydration by raw sample at the time of testing. The sample is introduced to the cassette via a transfer pipette simply by capillary force. DNA amplification was carried out in a portable prototype instrument for PCR thermal cycling with fluorescence detection of amplified products by melt curve analysis (MCA). To demonstrate performance, raw genital swabs and urine were introduced to the same cassette to simultaneously detect four sexually transmitted infections. Herpes Simplex Viruses (HSV-1 and HSV-2) were detected from raw genital swabs. Ureaplasma urealyticum (UU) and Mycoplasma homonis (MH) were detected from raw urine. Results for multiple patients were obtained in as little as 50 min. This platform allows multiparameter clinical testing with a pre-assembled cassette that requires only the introduction of raw sample. Modification of the prototype device to accommodate larger cassettes will ultimately provide high throughput simultaneous testing of even larger numbers of samples for many different targets, as is required for some clinical applications. Combinations of wax and/or polymer cassettes holding capillary reaction units are feasible. The components of the cassette are suited to mass production and robotic assembly to produce a readily manufactured disposable reaction cassette that can be configured for disease-specific testing panels. Rapid testing with a disposable reaction cassette on an inexpensive instrument will enable on the spot evaluation of patients in the clinic for faster medical decision-making and more informed therapeutic choices.

The early history of microwave spectroscopy is reviewed. New directions in the field are indicated. These include: further extensions of coherent submillimeter wave spectroscopy, microwave spectroscopy of molecules in interstellar space, microwave-infrared laser double resonance, spectroscopy of ionized molecules and transient molecular radicals, studies of hydrogen-bonded molecular complexes and atom-molecule complexes, observations of “forbidden” rotational transitions in symmetric-top and spherical-top molecules, and new developments in high-temperature spectroscopy.

Hyperfine structure due to (17)O has been measured in irradiated zein and edestin after exposure to gaseous oxygen with (17)O concentrated to 24%. The observations prove that the free radicals produced by ionizing irradiation under vacuum at 300 K are converted by the oxygen to protein-peroxide free radicals X-O((1))-O((2)), with the unpaired electron density in a pi-type orbital predominantly on the peroxide group. From the observed couplings, the 2p(pi) spin densities on zein-peroxide are found to be 0.29 and 0.45 for O((1)) and O((2)), respectively; those on edestin-peroxide are 0.26 and 0.48 for O((1)) and O((2)), respectively.

A millimeter-wave spectrometer having a sensitivity of 4 × 10−10 cm−1 in the 2-mm region has been used for observation of the “forbidden” transitions J → J, K = ±4 → ±1 and J → J, K = ±5 → ±2 in AsH3. A comprehensive computer analysis was made of the frequencies measured in this work together with available microwave frequencies of other transitions. This analysis provides accurate values of the rotational constants, nuclear quadrupole couplings, and effective structural parameters of the molecule. The spectral constants B0 and C0 (in MHz) are 112 470.597 and 104 884.665, respectively.

A millimeter-wave spectrometer having a sensitivity of 4 × 10−10 cm−1 in the 2-mm region has been constructed for observation of extremely weak millimeter-wave spectra of gases. It has been used to measure J → J, K = 0 ← 3 transitions in PH3 and J → J, K = 0 ← 3 as well as K = ±1 ← ±4 transitions in PD3. The B0 and C0 spectral constants (in MHz) are: for PH3, B0 = 133 480.15 ± 0.12 and C0 = 117 488.85 ± 0.16; for PD3, B0 = 69 471.10 ± 0.03 and C0 = 58 974.37 ± 0.05. The effective ground-state values obtained for the bond angle and bond length are: for PH3, r0 (Å) = 1.4200 and α0(o) = 93.345; for PD3, r0 (Å) = 1.4176 and α0(o) = 93.359. The corresponding zero-point-average values were calculated to be: for PH3, rz (Å) = 1.42699 ± 0.0002 and αz(o) = 93.2287; for PD3, rz (Å) = 1.42265 ± 0.0001 and αz(o) = 93.2567 ± 0.004. For both species, the equilibrium values are re (Å) = 1.41159 ± 0.0006 and αe(o) = 93.328 ± 0.02.

A new high-temperature millimeter-wave microwave spectrometer has been constructed and used to measure a wide range of vibrational and rotational states of silicon monoxide. This work results in accurate rest frequencies for all of the intersteller SiO maser transitions that have been observed as well as accurate measurements or predictions of all transitions that are likely to be of astrophysical interest. In addition, the Dunham spectral and potential constants are calculated for the three major isotopic species. For $^{28}\mathrm{Si}$$^{16}\mathrm{O}$: ${Y}_{01}=21787.453(11)$ MHz, ${Y}_{11}=\ensuremath{-}151.026(11)$ MHz, ${Y}_{21}=70.5(24)$ kHz, ${Y}_{02}=\ensuremath{-}29.38(13)$ kHz, ${a}_{0}=5.390(22)\ifmmode\times\else\texttimes\fi{}{10}^{5}$ ${\mathrm{cm}}^{\ensuremath{-}1}$, ${a}_{1}=\ensuremath{-}2.9899(41)$, ${a}_{2}=5.7(7)$, ${a}_{3}=\ensuremath{-}9.0(5)$. The calculated equilibrium parameters for $^{28}\mathrm{Si}$$^{16}\mathrm{O}$ are ${B}_{e}=21787.5(10)$ MHz, ${\ensuremath{\omega}}_{e}=1252.(3)$ ${\mathrm{cm}}^{\ensuremath{-}1}$, ${\ensuremath{\omega}}_{e}{x}_{e}=5.96(71)$ ${\mathrm{cm}}^{\ensuremath{-}1}$, and ${r}_{e}=1.50973(4)$ \AA{}.

Sodium atoms have been deposited on various pyrimidine powders under high vacuum at 77 degrees K and the electron spin resonance spectra of the resulting free radicals have been observed. Generally, the spectra show that the electron of the Na goes into a molecular orbital of the pyrimidine ring and the Na+ ions become attached to a carbonyl oxygen of the resulting pyrimidine anion. In 5-fluorouracil and 5-chlorouracil, however, the halogen is evidently abstracted by the Na to form NaF or NaC1 and the neutral uracil radical. Thymine shows evidence for H-addition radicals as well as the Na+-[thymine]-complex. The H source for the addition radicals may be an H-2-0 impurity in the sample, with which the Na atoms combine to release the H atoms. In addition to a resonance with g equals 2.00 from the pyrimidine anion radical, broad resonances with g greater than 2 were observed for 5-bromouracil, 5-chlorouracil and 5-iodouracil, as well as for alloxan and cytosine. These resonances, generally unstable at room temperature, are believed to arise from electrons trapped in interstitial sties or vacancies in the lattice.

Measurements of millimeter-wave fine-structure transitions and the $n(J)=0(1)\ensuremath{\rightarrow}2(1)$ rotational transition of $^{16}\mathrm{O}$ $^{18}\mathrm{O}$ have been made with high precision. From analysis of the results the following molecular constants of $^{16}\mathrm{O}$ $^{18}\mathrm{O}$ were obtained: ${B}_{0}=40707.408(10)$ MHz, ${B}_{1}=\ensuremath{-}0.129$ MHz, ${\ensuremath{\lambda}}_{0}=59499.097(43)$ MHz, ${\ensuremath{\lambda}}_{1}=0.05312(80)$ MHz, ${\ensuremath{\mu}}_{0}=\ensuremath{-}238.488(7)$ MHz, ${\ensuremath{\mu}}_{1}=\ensuremath{-}0.000619(116)$ MHz, ${B}_{e}=40931.7(6.9)$ MHz, ${r}_{0}=1.210751(16)$ \AA{}, and ${r}_{e}=1.207429(103)$ \AA{}. The equilibrium values were obtained with the aid of vibration-rotation interaction constants from optical spectroscopy.