Comparative study of three short-chain neurotoxins from the venom of Naja kaouthia (Yunnan, China)

Three short-chain neurotoxins named NT-I, NT-II, and NT-III were purified from the venom of Naja kaouthia, a snake distributed throughout the south of Yunnan province, China, by a series of chromatographic steps, including an FPLC Resource S column. Their molecular weights, determined by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS, were 6952.19 Da, 6854.92 Da, and 6828.80 Da, respectively. NT-I consisted of 62 amino acid residues, and the other two consisted of 61 amino acid residues, including 8 cysteines. After hydrolysis by endoproteinase Glu-C, their primary sequences were determined. A test of their activities demonstrated that they effectively inhibited muscle contractions induced by electric stimulation. Furthermore, the extent of inhibition caused by NT-II and NT-III was less than that of NT-I. The IC(50)s were 0.04 mug/ml, 0.20 mug/ml, and 0.23 mug/ml for NT-I, NT-II, and NT-III, respectively. Compared with NT-II and NT-III, the higher activity of NT-I may be a result of the amino acid residue substitution Ile36 to Arg36.

A novel short neurotoxin, cobrotoxin c, from monocellate cobra (Naja kaouthia) venom: isolation and purification, primary and secondary structure determination, and tertiary structure modeling

A novel short neurotoxin, cobrotoxin c (CBT C) was isolated from the venom of monocellate cobra (Naja kaouthia) using a combination of ion-exchange chromatography and FPLC. Its primary structure was determined by Edman degradation. CBT C is composed of 61 amino acid residues. It differs from cobrotoxin b (CBT B) by only two amino acid substitutions, Thr/Ala11 and Arg/Thr56, which are not located on the functionally important regions by sequence similarity. However, the LD50 is 0.08 mg/g to mice, i.e. approximately five-fold higher than for CBT B. Strikingly, a structure-function relationship analysis suggests the existence of a functionally important domain on the outside of Loop III of CBT C. The functionally important basic residues on the outside of Loop III might have a pairwise interaction with alpha subunit, instead of gamma or delta subunits of the nicotinic acetylcholine receptor (nAChR). (C) 2002 Elsevier Science Inc. All rights reserved.

Short Homologous Sequences Are Strongly Associated with the Generation of Chimeric RNAs in Eukaryotes

Chimeric RNAs have been reported in varieties of organisms and are conventionally thought to be produced by trans-splicing of two or more distinct transcripts. Here, we conducted a large-scale search for chimeric RNAs in the budding yeast, fruit fly, mous

A STUDY ON THE EFFECT OF LESIONS OF AREA-7 OF THE PARIETAL CORTEX ON THE SHORT-TERM VISUAL SPATIAL MEMORY OF RHESUS-MONKEYS (MACACA-MULATTA)

This research is focused on the contribution of area 7 to the short-term visual spatial memory. Three rhesus monkeys (Macaca mulatta) were trained in the direct delayed response task in which 5 delay intervals were used in each session. When each monkey reached the criterion of 90% correct responses in 5 successive sessions, two monkeys underwent a surgery while the other one received a sham operation as a control. In the first stage of the surgery, bilateral areas 7a, 7b and 7ip of the parietal cortex of two monkeys were precisely lesioned. After 7 days of recuperation, the monkeys were required to do the same task. The average percentage of correct responses in the lesioned animals decreased from 94.7% to 89.3% and 93.3% to 82.0% respectively (no significance, P > 0.05, n = 2). In addition, the monkeys' complex movements were mildly impaired. The lesioned monkeys were found to have difficulty picking up food from the wells. In the second stage, bilateral area 7m was lesioned. In the 5 postoperative sessions, the average percentage of correct responses in one monkey, with a relatively precise 7m lesion, decreased from 94.7% to 92.2% (no significance, P > 0.05), while the other monkey, with widely spread necrosis of lateral parietal cortex, showed an. obvious decline in performance, but still over the chance level. After 240 trials this monkey reattained the normal criterion. The results of this research suggest that the lesions of area 7 of the parietal cortex did not significantly affect the short-term visual spatial memory, which has been shown to be sensitive to lesions of the prefrontal cortex; they also support the notion of dissociation of spatial functions in the prefrontal and parietal cortices.

Glycine receptor activation regulates short-term plasticity in CA1 area of hippocampal slices of rats

Functional glycine receptors (GlyRs) are enriched in the hippocampus, but their roles in synaptic transmission are unclear. In this study, we examined the effect of GlyR activation on paired-pulse stimulation of the whole-cell postsynaptic currents (PSCs)

Short-term effects of drawing water for connectivity of rivers and lakes on zooplankton community structure

During 28-29, September 2005, water was drawn from Hanjiang River and Houguan Lake to the Yangzi River via Sanjiao Lake and Nantaizi Lake in Wuhan in order to provide favorable conditions for ecosystem restoration. To evaluate the feasibility and validity of drawing water as a means of ecosystem restoration, zooplankton populations were studied 3 times (before, immediately after finishing and a month after drawing water) at seven locations from 27 Sept. 2005 to 2 Nov. 2005. Water quality in the lakes was mostly improved and zooplankton species richness decreased as soon as drawing water had finished but increased a month after drawing water. Zooplankton density and biomass was reduced in the lakes by drawing water but was increased at the entrance to Sanjiao Lake because of landform geometry change. Before drawing water, most species in Sanjiao Lake e.g., Brachionus sp. and Keratella sp. were tolerant of contamination. After drawing water oligotrophic-prone species such as Lecane ludwigii and Gastropus stylifer emerged. We conclude that drawing water could be important for improving water quality and favour ecosystem restoration. Dilution of nutrient concentrations may be an important role in the effect.

The cytomegalovirus promoter-driven short hairpin RNA constructs mediate effective RNA interference in zebrafish in vivo

The ability to utilize the RNA interference (RNAi) machinery for silencing target-gene expression has created a lot of excitement in the research community. In the present study, we used a cytomegalovirus (CMV) promoter-driven DNA template approach to induce short hairpin RNA (shRNA) triggered RNAi to block exogenous Enhanced Green Fluorescent Protein (EGFP) and endogenous No Tail (NTL) gene expressions. We constructed three plasmids, pCMV-EGFP-CMV-shGFP-SV40, pCMV-EGFP-CMV-shNTL-SV40, and pCMV-EGFP-CMV-shScrambled-SV40, each containing a CMV promoter driving an EGFP reporter cDNA and DNA coding for one shRNA under the control of another CMV promoter. The three shRNA-generating plasmids and pCMV-EGFP control plasmid were introduced into zebrafish embryos by microinjection. Samples were collected at 48 h after injection. Results were evaluated by phenotype observation and real-time fluorescent quantitative reverse-transcription polymerase chain reaction (Q-PCR). The shGFP-generating plasmid significantly inhibited the EGFP expression viewed under fluorescent microscope and reduced by 70.05 +/- 1.26% of exogenous EGFP gene mRNA levels compared with controls by Q-PCR. The shRNA targeting endogenous NTL gene resulted in obvious NTL phenotype of 30 +/- 4% and decreased the level of their corresponding mRNAs up to 54.52 +/- 2.05% compared with nontargeting control shRNA. These data proved the feasibility of the CMV promoter-driven shRNA expression technique to be used to inhibit exogenous and endogenous gene expressions in zebrafish in vivo.

Knock down of gfp and no tail expression in zebrafish embryo by in vivo-transcribed short hairpin RNA with T7 plasmid system

A short-hairpin RNA (shRNA) expression system, based on T7 RNA polymerase (T7RP) directed transcription machinery, has been developed and used to generate a knock down effect in zebrafish embryos by targeting green fluorescent protein (gfp) and no tail (ntl) mRNA. The vector pCMVT7R harboring T7RP driven by CMV promoter was introduced into zebrafish embryos and the germline transmitted transgenic individuals were screened out for subsequent RNAi application. The shRNA transcription vectors pT7shRNA were constructed and validated by in vivo transcription assay. When pT7shGFP vector was injected into the transgenic embryos stably expressing T7RP, gfp relative expression level showed a decrease of 68% by analysis of fluorescence real time RT-PCR. As a control, injection of chemical synthesized siRNA resulted in expression level of 40% lower than the control when the injection dose was as high as 2 mu g/mu l. More importantly, injection of pT7shNTL vector in zebrafish embryos expressing T7RP led to partial absence of endogenous ntl transcripts in 30% of the injected embryos when detected by whole mount in situ hybridization. Herein, the T7 transcription system could be used to drive the expression of shRNA in zebrafish embryos and result in gene knock down effect, suggesting a potential role for its application in RNAi studies in zebrafish embryos.

Short and long peptidoglycan recognition proteins (PGRPs) in zebrafish, with findings of multiple PGRP homologs in teleost fish

Peptidoglycan recognition protein (PGRP) specifically binds to peptidoglycan and is considered to be one of the pattern recognition proteins in the innate immunity of insect and mammals. Using a database mining approach and RT-PCR, multiple peptidoglycan recognition protein (PGRP) like genes have been discovered in fish including zebrafish Danio rerio, Japanese pufferfish TakiFugu rubripes and spotted green pufferfish Tetraodon nigroviridis. They share the common features of those PGRPs in arthropod and mammals, by containing a conserved PGRP domain. Based on the predicted structures, the identified zebrafish PGRP homologs resemble short and long PGRP members in arthropod and mammals. The identified PGRP genes in T. nigroviridis and TakiFugu rubripes resemble the long PGRPs, and the short PGRP genes have not been found in T. nigroviridis and TakiFugu rubripes databases. Computer modelling of these molecules revealed the presence of three alpha-helices and five or six beta-strands in all fish PGRPs reported in the present study. The long PGRP in teleost fish have multiple alternatively spliced forms, and some of the identified spliced variants, e.g., tnPGRP-L3 and tnPGRP-L4 (in: Tetraodon nigroviridis), exhibited no characters present in the PGRP homologs domain. The coding regions of zfPGRP6 (zf: zebrafish), zfPGRP2-A, zfPGRP2-B and zfPGRP-L contain five exons and four introns; however, the other PGRP-like genes including zfPGRPSC1a, zfPGRPSC2, tnPGRP-L1-, tnPGRP-L2 and frPGRP-L (fr: Takifugu rubripes) contain four exons and three introns. In zebrafish, long and short PGRP genes identified are located in different chromosomes, and an unknown locus containing another long PGRP-like gene has also been found in zebrafish, demonstrating that multiple PGRP loci may be present in fish. In zebrafish, the constitutive expressions of zfPGRP-L, zfPGRP-6 and zfPGRP-SC during ontogeny from unfertilized eggs to larvae, in different organs of adult, and the inductive expression following stimulation by Flavobacterium columnare, were detected by real-time PCR, but the levels and patterns varied for different PGRP genes, implying that different short and long PGRPs may play different roles in innate immune response. (c) 2007 Elsevier Ltd. All rights reserved.

Multiplex detection of mutations in clinical isolates of rifampin-resistant Mycobacterium tuberculosis by short oligonucleotide Ligation assay on DNA chips

A new approach, short-oligonucleotide-ligation assay on DNA chip (SOLAC), is developed to detect mutations in rifampin-resistant Mycobacterium tuberculosis. The method needs only four common probes to detect 15 mutational variants of the rpoB gene within 12 h. Fifty-five rifampin-resistant M. tuberculosis isolates were analyzed, resulting in 87.3% accuracy and 83.6% concordance relative to DNA sequencing.

High repetition rate Q-switched microchip Nd:YVO4 laser with pulse duration as short as 1.1 ns

We have demonstrated a compact and an efficient passively Q-switched microchip Nd:YVO4 laser by using a composite semiconductor absorber as well as an output coupler. The composite semiconductor absorber was composed of an LT (low-temperature grown) In0.25Ga0.75As absorber and a pure GaAs absorber. To our knowledge, it was the first demonstration of the special absorber for Q-switching operation of microchip lasers. Laser pulses with durations of 1.1 ns were generated with a 350 mu m thick laser crystal and the repetition rate of the pulses was as high as 4.6 MHz. The average output power was 120 mW at the pump power of 700 mW. Pulse duration can be varied from 1.1 to 15.7 ns by changing the cavity length from 0.45 to 5 mm. Pulses with duration of 1.67 and 2.41 ns were also obtained with a 0.7 mm, thick laser crystal and a 1 mm thick laser crystal, respectively. (C) 2007 Elsevier GmbH. All rights reserved.

InAs/GaSb superlattices for photodetection in short wavelength infrared range

The first report of a short wavelength infrared detector based on type II InAs/GaSb superlattices is presented. Very short period superlattices containing InAs (2ML)/GaSb (8ML) superlattices (SLs) were grown by molecular-beam epitaxy on GaSb substrates. The photoluminescence showed a cut-off wavelength at 2.1 mu m at 10 K and 2.6 mu m at 300 K. Room-temperature optical transmittance spectra shows obvious absorption in InAs (2ML)/GaSb (8ML) SL in the range of 450-680 meV, i.e. 1.8-2.7 mu m. The cut-off wavelength moved from 2.3 mu m to 2.6 mu m with temperature rising from 77 K to 300 K in photoresponse spectra. The blackbody response R-v exponentially decreased as a function of 1/T in two temperature sections (130-200 K and 230-300 K). The blackbody detectivity D-bb(center dot) was beyond 1 x 10(8) cmHz(1/2)/W at room temperature. (C) 2009 Elsevier B.V. All rights reserved.

Transient Reorientation of a Doped Liquid Crystal System under a Short Laser Pulse

The transient optical nonlinearity of a nematic liquid crystal doped with azo-dye DR19 is examined. The optical reorientation threshold of a 25-mu m-thick planar-aligned sample of 5CB using a 50 ns pulse duration 532 nm YAG laser pulse is observed to decrease from 800 mJ/mm(2) to 0.6 mJ/mm(2) after the addition of 1 vol% azo dopant, a reduction of three orders of magnitude. When using a laser pulse duration of 10 ns, no such effect is observed. Experimental results indicate that the azo dopant molecules undergo photoisomerization from trans-isomer to cis-isomer under exposure to light, and this conformation change reorients the 5CB molecules via intermolecular coupling between guest and host. This guest-host coupling also affects the azo photoisomerization process.

SHORT PERIOD InAs/GaSb SUPERLATTICE INFRARED DETECTOR ON GaAs SUBSTRATES

Two type II superlattices (SLs) InAs(2ML)/GaSb(8ML) and InAs(8ML)/GaSb(8ML) were grown on GaAs substrates by molecular-beam epitaxy. High resolution X-ray diffraction showed the periods of the two SLs were 31.2 angstrom and 57.3 angstrom, respectively. Room-temperature optical transmittance spectra showed that there were clear absorption edges at 2.1 mu m and 5 mu m for the two SLs. The SWIR and MWIR photoconductor devices were fabricated by standard lithography and etched by tartaric acid solution. The spectral response and blackbody tests were carried out at low and room temperatues. The results show that the 50% cutoff wavelengths of the two photoconductors are 2.1 mu m and 5.0 mu m respectively and D-bb* is above 2 x 10(8) cmHz(1/2)/W for two kinds of photoconductors at 77K. D-bb* is above 10(8) cmHz(1/2)/W for SWIR photoconductor at room temperature.

GaAs Based InAs/GaSb Superlattice Short Wavelength Infrared Detectors Grown by Molecular Beam Epitaxy

InAs/GaSb superlattice (SL) short wavelength infrared photoconduction detectors are grown by molecular beam epitaxy on GaAs(001) semi-insulating substrates. An interfacial misfit mode AlSb quantum dot layer and a thick GaSb layer are grown as buffer layers. The detectors containing a 200-period 2ML/8ML InAs/GaSb SL active layer are fabricated with a pixel area of 800 x 800 mu m(2) without using passivation or antireflection coatings. Corresponding to the 50% cutoff wavelengths of 2.05 mu m at 77K and 2.25 mu m at 300 K, the peak detectivities of the detectors are 4 x 10(9) cm.Hz(1/2)/W at 77K and 2 x 10(8) cm.Hz(1/2)/W at 300 K, respectively.