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Developing fiber specific promoter-reporter transgenic lines to study the effect of abiotic stresses on fiber development in cotton. - PloS one
Cotton is one of the most important cash crops in US agricultural industry. Environmental stresses, such as drought, high temperature and combination of both, not only reduce the overall growth of cotton plants, but also greatly decrease cotton lint yield and fiber quality. The impact of environmental stresses on fiber development is poorly understood due to technical difficulties associated with the study of developing fiber tissues and lack of genetic materials to study fiber development. To address this important question and provide the need for scientific community, we have generated transgenic cotton lines harboring cotton fiber specific promoter (CFSP)-reporter constructs from six cotton fiber specific genes (Expansin, E6, Rac13, CelA1, LTP, and Fb late), representing genes that are expressed at different stages of fiber development. Individual CFSP::GUS or CFSP::GFP construct was introduced into Coker 312 via Agrobacterium mediated transformation. Transgenic cotton lines were evaluated phenotypically and screened for the presence of selectable marker, reporter gene expression, and insertion numbers. Quantitative analysis showed that the patterns of GUS reporter gene activity during fiber development in transgenic cotton lines were similar to those of the native genes. Greenhouse drought and heat stress study showed a correlation between the decrease in promoter activities and decrease in fiber length, increase in micronaire and changes in other fiber quality traits in transgenic lines grown under stressed condition. These newly developed materials provide new molecular tools for studying the effects of abiotic stresses on fiber development and may be used in study of cotton fiber development genes and eventually in the genetic manipulation of fiber quality.
[Effects of intra-periaqueductal gray injection of zeta pseudosubstrate inhibitory peptide on sensory and affective components of pain]. - Zhonghua yi xue za zhi
To explore the effects of intra-PAG injection of ZIP on sensory and affective components of pain.For determining the role of ZIP on pain-induced aversion, the effects of intra-PAG injection of ZIP on formalin-induced conditioned place avoidance (F-CPA) was investigated. To determine the role of ZIP on pain perception, formalin-induced inflammatory pain model was established and the effects of intra-PAG injection of ZIP on formalin-induced nociceptive behaviors was investigated.In the NS-treated rats, the time in the pain-paired compartment during the test session was significantly shorter than that during the preconditioning session 2 and 24 hours after administration of the drug at both 1 and 7 day post-training (Group NS-1 d-2 h: (465.1 Â± 40.6) vs (133.8 Â± 29.4) s (P < 0.001); Group NS-7 d-2 h: (432.3 Â± 43.7) vs (150.5 Â± 26.6) s (P < 0.01); Group NS-1 d-24 h: (500.5 Â± 20.6) vs (107.0 Â± 15.7) s (P < 0.001); Group NS-7 d-24 h: (450.8 Â± 27.4) vs (129.4 Â± 21.1) s (P < 0.001)). On the contrary, in the ZIP-treated rats, no significant differences were observed in the time in the pain-paired compartment between the post-conditioning and pre-conditioning sessions at the same time-points. CPA scores also showed the attenuation of F-CPA by intra-PAG injection of ZIP in comparison to the saline-injected rats (P < 0.05). Compared with the intra-PAG saline-injected group, intra-PAG microinjection of ZIP did not affect the formalin-induced nociceptive behaviors (P > 0.05).The study suggests that PAG contributes to pain-related aversion in rats, and the mechanism of pain emotion encoding in PAG may attribute to the activation of targets of ZIP.
Enhancement of reproductive heat tolerance in plants. - PloS one
Comparison of average crop yields with reported record yields has shown that major crops exhibit annual average yields three- to seven-fold lower than record yields because of unfavorable environments. The current study investigated the enhancement of pollen heat tolerance through expressing an Arabidopsis thaliana heat shock protein 101 (AtHSP101) that is not normally expressed in pollen but reported to play a crucial role in vegetative thermotolerance. The AtHSP101 construct under the control of the constitutive ocs/mas 'superpromoter' was transformed into cotton Coker 312 and tobacco SRI lines via Agrobacterium mediated transformation. Thermotolerance of pollen was evaluated by in vitro pollen germination studies. Comparing with those of wild type and transgenic null lines, pollen from AtHSP101 transgenic tobacco and cotton lines exhibited significantly higher germination rate and much greater pollen tube elongation under elevated temperatures or after a heat exposure. In addition, significant increases in boll set and seed numbers were also observed in transgenic cotton lines exposed to elevated day and night temperatures in both greenhouse and field studies. The results of this study suggest that enhancing heat tolerance of reproductive tissues in plant holds promise in the development of crops with improved yield production and yield sustainability in unfavorable environments.
[Effects of volume-controlled ventilation and pressure-controlled volume- guaranteed mode during one-lung ventilation on circulation, pulmonary function and lung injury]. - Zhonghua yi xue za zhi
To compare the effects of volume-controlled ventilation (VCV) and pressure-controlled volume-guaranteed (PCV-VG) mode during one-lung ventilation (OLV) on circulation, pulmonary function and lung injury.2012 February to 2013 March in Ningbo No2. Hospital cardiothoracic surgery, 30 patients aged 52 to 76 years (ASA grade II-III) undergoing elective thoracoscopic lobectomy were randomly divided into VCV group and PCV-VG group, with 15 cases in each group. After anesthesia induction and endotracheal intubation, endobronchial blocker was inserted to start OLV. Heart rate (HR), mean arterial pressure (MAP), measured tidal volume (TV), peak airway pressure (Ppeak), airway resistance (Raw), chest compliance (Cdyn) and the end-tidal carbon dioxide pressure (PetCO(2)) were recorded at the time point of 15 minutes after turning to the lateral position, 15 minutes and 60 minutes after OLV, and 15 minutes after the resumption of two lung ventilation. In the meanwhile, arterial blood gas analysis was conducted to measure indicators of pH, oxygen tension (PaO(2)) and carbon dioxide partial pressure (PaCO(2)). Blood was drawn before induction, 1 hour after OLV and 1 hour after the end of surgery, and the concentration of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-Î±) were detected by enzyme-linked immunosorbent assay (ELISA).HR, MAP, TV, PetCO(2), pH and PaCO(2) in two groups at the time point of 15 minutes after turning to the lateral position, 15 minutes and 60 minutes after OLV, and 15 minutes after the resumption of two lung ventilation showed no significant difference (P > 0.05). At the point of 15 minutes after turning to the lateral position and 15 minutes after two lung ventilation, Ppeak and Cdyn of two groups were significantly different (P < 0.05) (Ppeak:16 Â± 3 cmH(2)O, 16 Â± 3 cmH(2)O for VCV group and 14 Â± 2 cmH(2)O, 14 Â± 2 cmH(2)O for PCV-VG group; Cdyn: 43.5 Â± 5.9 ml/cmH(2)O, 43.8 Â± 6.7 ml/cmH2O for VCV group and 49.7 Â± 7.1 ml/cmH(2)O, 53.3 Â± 9.6 ml/cmH(2)O for PCV-VG group). Compared with VCV group, PCV-VG group showed a lower Ppeak 15 minutes and 60 minutes after OLV [ (17 Â± 2 cmH(2)O) vs (22 Â± 4 cmH(2)O) and (18 Â± 4 cmH(2)O) vs( 23 Â± 3 cmH(2)O) with a higher Cdyn at the same point (38.6 Â± 6.3 ml/cmH(2)O) vs (29.6 Â± 3.2 ml/cmH(2)O) and 37.3 Â± 6.0 ml/cmH(2)O) vs (30.3 Â± 3.8 ml/cmH(2)O)] (P < 0.01). Compared with VCV group,IL-6 and TNF-Î± of PCV-VG group 1 hour after OLV and 1 hour after the end of surgery were significantly lower (P < 0.01) (IL-6: 52.32 Â± 3.59 vs 62.65 Â± 4.17 pg/ml and 63.57 Â± 4.98 vs 82.38 Â± 4.10 pg/ml; TNF-Î±: 3.23 Â± 0.27 vs 4.19 Â± 0.38 pg/ml and 4.01 Â± 0.28 vs. 5.49 Â± 0.31 pg/ml).During one-lung ventilation in thoracoscopic lobectomy, PCV-VG mode has a competitive advantage over VCV mode in terms of pulmonary function and lung protection.
Immune complex formation in human diabetic retina enhances toxicity of oxidized LDL towards retinal capillary pericytes. - Journal of lipid research
Recently it has been shown that levels of circulating oxidized LDL immune complexes (ox-LDL-ICs) predict the development of diabetic retinopathy (DR). This study aimed to investigate whether ox-LDL-ICs are actually present in the diabetic retina, and to define their effects on human retinal pericytes versus ox-LDL. In retinal sections from people with type 2 diabetes, costaining for ox-LDL and IgG was present, proportionate to DR severity, and detectable even in the absence of clinical DR. In contrast, no such staining was observed in retinas from nondiabetic subjects. In vitro, human retinal pericytes were treated with native LDL, ox-LDL, and ox-LDL-IC (0-200 mg protein/l), and measures of viability, receptor expression, apoptosis, endoplasmic reticulum (ER) and oxidative stresses, and cytokine secretion were evaluated. Ox-LDL-IC exhibited greater cytotoxicity than ox-LDL toward retinal pericytes. Acting through the scavenger (CD36) and IgG (CD64) receptors, low concentrations of ox-LDL-IC triggered apoptosis mediated by oxidative and ER stresses, and enhanced inflammatory cytokine secretion. The data suggest that IC formation in the diabetic retina enhances the injurious effects of ox-LDL. These findings offer new insights into pathogenic mechanisms of DR, and may lead to new preventive measures and treatments.
Adductor canal block can result in motor block of the quadriceps muscle. - Regional anesthesia and pain medicine
The block of nerves in the adductor canal is considered to cause a sensory block without a motor component. In this report, we describe a case of significant quadriceps muscle weakness after an adductor canal block (ACB).A 65-year-old female patient for ambulatory knee surgery was given an ACB for postoperative pain management. The block was performed under ultrasound guidance at the midthigh level using the transsartorial approach. Twenty milliliters of 0.5% ropivacaine was deposited adjacent to the anterior and posterior areas of the femoral artery. On discharge from the hospital, the patient realized that her thigh muscles were weak and she was unable to extend her leg at the knee. A neuromuscular examination indicated that the patient had no strength in her quadriceps muscle, along with sensory deficit in the medial-anterior lower leg and area in front of knee up to the midthigh. The weakness lasted 20 hours, and the sensory block lasted 48 hours before complete recovery. The optimal level and amount of local anesthetic for adductor canal block are currently not well defined. Proximal spread of local anesthetic and anatomical variation may explain our observation.Several studies have reported that ACB involves no motor blockade. However, our case report illustrates that the ACB can result in clinically significant quadriceps muscle paralysis. This report suggests that patients should be monitored vigilantly for this occurrence to decrease the risk of falls.
Continuous interscalene block in patients having outpatient rotator cuff repair surgery: a prospective randomized trial. - Anesthesia and analgesia
We performed this randomized trial to compare the recovery profile of patients receiving single injection (SISB) and continuous interscalene brachial plexus block (CISB) or general anesthesia (GA) for arthroscopic rotator cuff repair surgery through the first postoperative week. Our primary hypothesis was that the highest pain numeric rating scale (NRS) (worst pain score) at the end of the study week would be lower for patients in the CISB group than for patients in the SISB or GA groups.Seventy-one patients scheduled for elective outpatient arthroscopic rotator cuff repair were enrolled. CISB patients received 20 mL of 0.5% ropivacaine as a bolus through a catheter, whereas SISB patients received the same injection volume through a needle. CISB patients received an infusion of 0.2% ropivacaine at 5 mL/h with a patient-controlled bolus of 5 mL hourly for 48 hours. GA-only patients received a standardized general anesthetic. Postoperative highest NRS pain scores through the first postoperative week, time-to-first pain, analgesic consumption, fast-tracked postoperative anesthesia care unit (PACU) bypass rate, length of PACU stay, time-to-discharge home, total hours of sleep, and related adverse effects were recorded in the PACU and at home on postoperative days 1, 2, 3, and 7.No patient in the CISB or SISB groups reported a NRS â‰¥1 or required analgesics while in the PACU. While most patients in the CISB and SISB groups were fast-tracked to PACU discharge, no patient in the GA group was fast-tracked (Î§ P = 0.003). Length of stay in the PACU was significantly shorter for the CISB and SISB groups than for the GA group (20 Â± 31, 30 Â± 42, and 165 Â± 118 minutes, respectively (CISB vs GA, P < 0.001; SISB vs GA, P <0.001), and time-to-discharge home was significantly shorter when compared with the GA group. Time to first pain report was longer in the CISB group. Mean NRS scores were lower for patients in the CISB group than in the SISB and GA groups on postoperative days 1 and 2, and use of narcotics (doses â‰¥1) was lower until postoperative day 3. Patients who received CISB slept significantly longer than patients who received SISB or GA (P < 0.01) during the first 48 hours postoperatively. By the end of the study week, 26% of patients in the CISB group, 83% in the SISB group, and 58% of GA patients reported NRS â‰¥4 (both P-values â‰¤ 0.05).The analgesic benefits of CISB found in the PACU and immediately after discharge extend through the intermediate recovery period ending on postoperative day 7.
High-definition ultrasound imaging defines the paraneural sheath and the fascial compartments surrounding the sciatic nerve at the popliteal fossa. - Regional anesthesia and pain medicine
The connective tissue layers that surround the sciatic nerve at the popliteal fossa are poorly defined. We present high-definition ultrasound images of the sciatic nerve, which were acquired during ultrasound-guided popliteal sciatic nerve block (SNB), that clearly demonstrate these fascial layers.Four patients undergoing hallux valgus surgery received an ultrasound-guided popliteal SNB using a high-definition ultrasound system. In the ultrasound images, the paraneural sheath was identified as a hyperechoeic fascial layer between the outer surface of the sciatic nerve (epineurium) and the epimysium of the surrounding muscles. The paraneural sheath was distinct from the epineurium, better delineated after the local anesthetic injection, and enveloped not only the sciatic nerve but also the common peroneal and tibial nerves separately. In the postblock sonograms, the local anesthetic was compartmentalized into 2 broad areas, that is, external (subepimyseal) and internal (subparaneural) to the paraneural sheath. The popliteal SNB was effective for surgical anesthesia in all 4 patients.We have demonstrated the paraneural sheath and the fascial compartments, that is, the "subepimyseal perineural compartment" and the "subparaneural compartment" that surround the sciatic nerve and act as conduits for local anesthetic spread during a popliteal SNB.
Satellite- and epoch differenced precise point positioning based on a regional augmentation network. - Sensors (Basel, Switzerland)
Precise Point Positioning (PPP) has been demonstrated as a simple and effective approach for user positioning. The key issue in PPP is how to shorten convergence time and improve positioning efficiency. Recent researches mainly focus on the ambiguity resolution by correcting residual phase errors at a single station. The success of this approach (referred to hereafter as NORM-PPP) is subject to how rapidly one can fix wide-lane and narrow-lane ambiguities to achieve the first ambiguity-fixed solution. The convergence time of NORM-PPP is receiver type dependent, and normally takes 15-20 min. Different from the general algorithm and theory by which the float ambiguities are estimated and the integer ambiguities are fixed, we concentrate on a differential PPP approach: the satellite- and epoch differenced (SDED) approach. In general, the SDED approach eliminates receiver clocks and ambiguity parameters and thus avoids the complicated residual phase modeling procedure. As a further development of the SDED approach, we use a regional augmentation network to derive tropospheric delay and remaining un-modeled errors at user sites. By adding these corrections and applying the Robust estimation, the weak mathematic properties due to the ED operation is much improved. Implementing this new approach, we need only two epochs of data to achieve PPP positioning converging to centimeter-positioning accuracy. Using seven days of GPS data at six CORS stations in Shanghai, we demonstrate the success rate, defined as the case when three directions converging to desired positioning accuracy of 10 cm, reaches 100% when the interval between the two epochs is longer than 15 min. Comparing the results of 15 min' interval to that of 10 min', it is observed that the position RMS improves from 2.47, 3.95, 5.78 cm to 2.21, 3.93, 4.90 cm in the North, East and Up directions, respectively. Combining the SDED coordinates at the starting point and the ED relative coordinates thereafter, we demonstrate the performance of RTK PPP with standard deviation of 0.80, 1.34, 0.97 cm in the North, East and Up directions.
A high throughput DNA extraction method with high yield and quality. - Plant methods
Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L.) Moench] leaves and dry seeds with high yield, high quality, and affordable cost.We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles.A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need.
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