Expression of AgTRPA1 in larval tissues.

<p>cDNA libraries from larval antennae, heads and bodies were generated by extracting mRNA followed by <i>in intro</i> reverse transcription. <i>rps7</i> and <i>agtrpa1</i> were amplified using gene-specific primers and run on a 2% agarose gel. “+” or “-“ indicates the presence or absence of reverse transcriptase, respectively.</p

Larval antenna is a peripheral thermosensory organ.

<p><b>a</b>) Arithmetic means ± S.E.M of total distance travelled in 300s for individual larva recorded from larvae lacking either antennae or maxillary palp were plotted (n≥12). Asterisks suggest <i>p</i><0.05 using Mann–Whitney <i>U</i> test to compare antennal ablation to sham ablation treatment. Kruskal-Wallis one-way analysis of variance was also utilized to compare larval mobility at all 5 temperatures following antennal ablation with <i>p</i>>0.05, indicating larvae without antennae were not capable of eliciting differential mobility at varying ambient temperatures comparing to sham treatment. <b>b</b>–<b>k</b>) Localization of AgTRPA1 mRNA was detected by fluorescent in situ hybridization (FISH). White arrow indicates localization of AgTRPA1 mRNA while green labels neuronal axons and dendrites. <b>l</b>–<b>o</b>) Red fluorescence indicates AgTRPA1 mRNA while green indicates the localization of AgOrco protein that is expressed in all ORNs. White arrow indicates AgTRPA1-expressing neuronal cell bodies while hollow arrow shows cluster of ORNs (Scale bar, 25µm).</p

Knockdown of AgTRPA1 mRNA via RNAi.

<p>Means of cycle threshold (CT) values for amplification of <i>agtrpa1</i> and <i>rps7</i> were shown (n=2). Quantitative RT-PCR was performed on cDNA isolated from whole larvae receiving AgTRPA1, non-specific siRNA and buffer injection. Relative mRNA abundance + S.E.M was plotted with data normalized to non-specific siRNA treatment using PFAFFL method.</p

Thermal preferences of WT 4^th instar An.

<div><p><b><i>gambiae</i> larvae following the shift of cultivation</b>. </p> <p><b>a</b>) A stack of larval trajectories (n≥10) recorded in 7 different thermal gradients were shown for larvae reared at 30° C. <b>b</b>) Larval thermotactic indices ± S.E.M were plotted for larvae reared at 30° C. Mann-Whitney <i>U</i> test was used to compare thermotactic indices at 30 and 36°C with a <i>p</i> value > 0.05.</p></div

AgTRPA1 mediates larval behavior within the shifted hot range.

<p><b>a</b>) Arithmetic means ± S.E.M of total distance travelled in 300s for injected larvae reared at 30° C were plotted. Asterisks indicate <i>p</i><0.05 comparing AgTRAP1 and Non-specific siRNA-injected larvae (Mann–Whitney <i>U</i> test). <b>b</b>) Stack of larval trajectories (n≥10) recorded in 31-41°C gradient for buffer and AgTRPA1, Non-specific siRNA treatments were shown. Thermotactic indices ± S.E.M were shown for injected larvae reared at 30° C. Asterisks indicate <i>p</i><0.05 comparing AgTRPA1 and Non-specific siRNA-injected larvae (Mann–Whitney <i>U</i> test).</p

Thermal-induced larval mobility following the shift of cultivation.

<p>Arithmetic means ± S.E.M recorded from larvae reared at both 27 and 30°C of total distance travelled in 300s were plotted (n≥12). White arrow shows the shift of cultivation temperature from 27 to 30°C. Black circle shows the temperature at which larval mortality was evident for both 27 and 30°C-reared colony. This figure indicates the change of larval mobility pattern matches the shift of rearing temperature.</p

Thermal-induced mobility in WT 4^th instar <i>An</i>.

<div><p><b><i>gambiae</i> larvae</b>. </p> <p>Arithmetic means ± standard error of the mean (S.E.M) of total distance travelled by individual larva in 300s were plotted (n≥15). Red circle indicates the two individual temperatures that generated lowest larval mobility in the neighboring temperature ranges (27 and 33°C) while black circle shows the temperature at which larvae experienced morbidity/death after 2-3 mins of assaying (41° C), thus the total distance was calculated based on the time frame before larval mortality.</p></div

CercaTest Red^TM, a novel urine-based point-of-care test for the detection of preeclampsia

Preeclampsia (PE) poses a serious threat to the health of the pregnant woman and her developing fetus due to the difficulty in diagnosing the condition. The disease can develop and worsen suddenly without noticeable signs and symptoms. Thus, there is an urgent need for a simple Point of Care Test (POCT) that improves accessibility to testing and can be used as an aid in the diagnosis of PE. CercaTest RedTM is a noninvasive detection kit for impending preeclampsia using urine from pregnant women. This is especially pertinent for women who have limited access to secondary/tertiary healthcare as those in remote settings, low-income countries or simply lack of out of hours laboratory services. The kit employs an absorptive column that separates Congo red dye bound to urinary misfolded protein from pregnant women and unbound dye. When a solution of Congo red dye pre-mixed with urine is loaded onto the absorptive matrix in a detection cuvette, the presence (positive) or absence (negative) of misfolded proteins can be determined based on the color of eluate collected in the lower section of the cuvette. 190 and 937 pregnant women who were >18 years old at the gestational age of ≥20 weeks were enrolled for the feasibility and validation cohort, respectively. The consistency between CercaTest RedTM and clinical diagnosis of PE according to the American College of Obstetricians and Gynecologist (ACOG) Guidelines was analyzed using the kappa statistic. The POCT has a limit of detection (LoD) of human urinary misfolded proteins equivalent to 0.45 μM of denatured human serum albumin, with high reproducibility and stability. An accuracy of 96.84% for diagnosis of preeclampsia with a Kappa statistic of 0.746 (p This test is easy to use, cost-effective and portable with short turnaround time and no laboratory instrument requirement. In the future, the test may have the potential to become quantitative using spectroscopy. (Chinese Clinical Trial Registry No. ChiCTR1800017692).</p

Supplemental Material - Influence of learning-space enclosure on attention in undergraduate students measured by eye-tracking technology

Supplemental Material for Influence of learning-space enclosure on attention in undergraduate students measured by eye-tracking technology by Chao Liu, Xiaotong Jing, Yalin Zhang and Weijun Gao in Indoor and Built Environment</p

Additional file 1 of Novel KMT2B gene mutation in MUC4 positive low-grade fibromyxoid sarcoma

Supplementary Material