Identification and Functional Characterization of cis-Regulatory Elements of Key Photorespiratory Genes in Response to Short-Term Abiotic Stress Conditions.

The cis-regulatory elements (CREs) are the short stretches of noncoding DNA upstream of a gene, which play a critical role in fine-tuning gene expression. Photorespiration is a multi-organellar, energy-expensive biochemical process that remains intricately linked to photosynthesis and is conserved in plants. Recently, much focus has been devoted in generating plants with engineered alternative photorespiratory bypasses to enhance photosynthetic efficiency without compromising the beneficial aspect of photorespiration. Varied constitutive or inducible promoters for generating transgenic plants harboring multiple transgenes have been introduced over years; however, most of them suffer from unintended effects. Consequently, a demand for synthetic tunable promoters based on canonical CRE signatures derived from native genes is on the rise. Here, in this chapter, we have provided a detailed method for in silico identification and characterization of CREs associated with photorespiration. In addition to the detailed protocol, we have presented an example of a typical result and explained the significance of the result. Specifically, the method covers how to identify and generate tunable synthetic promoters based on native CREs using three key photorespiratory genes from Arabidopsis and two web-based tools, namely, PlantPAN3.0 and AthaMap. Finally, we have also furnished a protocol on how to test the efficacies of the synthetic promoters harboring predicted CREs using transient tobacco expression coupled with luciferase-based promoter assay in response to ambient conditions and under short-term abiotic stress conditions.

Is Response to a Pre-implant Diagnostic Peripheral Nerve Block Associated With Efficacy After Peripheral Nerve Stimulation Implantation? A Ten-Year Enterprise-Wide Analysis.

OBJECTIVES: This study aimed to assess whether patient response to targeted diagnostic peripheral nerve block before peripheral nerve stimulator (PNS) device implantation is associated with efficacy after PNS implantation. MATERIALS AND METHODS: The electronic medical records from the Mayo Clinic Enterprise (three quarternary care medical centers and additional satellite medical centers) were reviewed to identify patients who underwent PNS implantation between January 2014 and January 2023. A primary outcome of interest was to assess whether administration of a preimplant diagnostic peripheral nerve block predicted pain relief at three months and six months after temporary and permanent PNS implantation. Another primary outcome was to investigate whether there was an association between the pain relief from a preimplant diagnostic peripheral nerve block and pain relief after three and six months after temporary or permanent PNS therapy. Linear regression analysis was conducted for outcomes of interest. RESULTS: Of 193 eligible patients who underwent PNS therapy, a total of 173 patients were included in the final analysis and were stratified into either the temporary PNS cohort (n = 112) or the permanent PNS cohort (n = 61). Overall, 77.5% of all patients (134/173) underwent a preimplant diagnostic peripheral nerve block and reported a mean percentage relief of 70.1 ± 27.0 from the diagnostic block. Of patients in the temporary PNS cohort, there was no difference in postimplant percentage pain relief between patients who received a diagnostic block (n = 93) and control patients (n = 19) at three months (35.4 ± 36.0 vs 49.8 ± 36.1, respectively; ß -14.45, 95% CI -32.98 to 4.07, p = 0.125) or at six months (23.3 ± 30.8 vs 45.7 ± 40.0, respectively; ß -22.39, 95% CI -46.86 to 2.08, p = 0.072). Of patients in the permanent PNS cohort, there was no difference in postimplant percentage pain relief between patients who received a diagnostic block (n = 41) and control patients (n = 20) at three months (42.4 ± 34.3 vs 43.2 ± 42.4, respectively; ß -0.79, 95% CI -23.56 to 21.99, p = 0.945) or at six months (44.3 ± 35.0 vs 38.8 ± 40.9, respectively; ß 5.42, 95% CI -20.04 to 30.88, p = 0.669). Pain relief from preimplant diagnostic blocks was associated with postimplant pain relief from temporary PNS at three months (ß 0.33, 95% CI 0.04-0.61, p = 0.025). However, pain relief from preimplant diagnostic blocks did not predict postimplant pain relief from temporary PNS at six months, or permanent PNS at three months and six months. CONCLUSIONS: Administration of a diagnostic block is not associated with superior pain relief at three or six months after PNS implantation to that of an approach without diagnostic block. Pain relief from a diagnostic block may potentially predict short-term pain relief after temporary PNS therapy, although future prospective studies are warranted to evaluate the prognostic utility of diagnostic blocks.

Adverse Events Associated With 10-kHz Dorsal Column Spinal Cord Stimulation: A 5-Year Analysis of the Manufacturer and User Facility Device Experience (MAUDE) Database.

BACKGROUND: High-frequency (10-kHz) spinal cord stimulation (SCS) continues to be an emerging therapy in chronic pain management. The same complications that plagued earlier SCS systems may affect newer stimulation technologies, although there is limited data on the type of complications and surgical management of these complications. OBJECTIVE: The aim of this study was to systematically examine real-world complications associated with 10-kHz SCS reported on the Manufacturer and User Facility Device Experience (MAUDE) database. MATERIALS AND METHODS: The MAUDE database was queried for entries reported between January 1, 2016 and December 31, 2020. Entries were classified into procedural complications, device-related complications, patient complaints, surgically managed complications, serious adverse events, and/or other complications. Primary outcomes included type and frequency of complications, and surgical management of complications. RESULTS: A total of 1651 entries were analyzed. Most entries were categorized as procedural complications (72.6%), followed by serious adverse events (10.5%), device-related complications (10.5%), and patient complaints (9.9%). Most complications were managed surgically with explant (50.9%) rather than revision (5.0%) or incision/drainage (6.6%). Of procedural complications, the most common entries included non-neuraxial infection (52.9%), new neurological symptoms (14.7%), and dural puncture (9.5%). Of device-related complications, the most common entries included lead damage (41.6%), erosion (18.5%), and difficult insertion (11.5%). CONCLUSION: This retrospective 5-year analysis of complications from10-kHz SCS provides a real-world assessment of safety data unique for this stimulation modality. This analysis may help inform future clinical decisions, lead to device enhancement and optimization, and improve mitigation of risks to provide safe and efficacious use of 10-kHz SCS.

Marine debris knows no boundaries: Characteristics of debris accumulation in marine protected areas of the Florida Keys.

Marine debris is a persistent and pervasive threat worldwide including inside marine protected areas (MPAs). To assess marine debris accumulation rates and potential impacts, we counted and evaluated trap, non-trap fishing gear, and non-fishing debris in unprotected areas and MPAs with different management boundary regulations in the Florida Keys (USA). Analyses identified that neither MPA type nor size were strong drivers of debris density and that debris densities were not statistically different between unprotected areas and MPAs. Non-fishing and non-trap fishing gear debris densities were potentially related to unexplored local differences in human behavior, while trap debris density was likely associated with oceanographic forces that transported traps into the MPAs. Overall, our results suggested that the drivers of marine debris accumulation for each debris category were different and may vary with each individual MPA, and that marine debris is not constrained by MPA boundaries.

Ocean acidification disrupts the orientation of postlarval Caribbean spiny lobsters.

Anthropogenic inputs into coastal ecosystems are causing more frequent environmental fluctuations and reducing seawater pH. One such ecosystem is Florida Bay, an important nursery for the Caribbean spiny lobster, Panulirus argus. Although adult crustaceans are often resilient to reduced seawater pH, earlier ontogenetic stages can be physiologically limited in their tolerance to ocean acidification on shorter time scales. We used a Y-maze chamber to test whether reduced-pH seawater altered the orientation of spiny lobster pueruli toward chemical cues produced by Laurencia spp. macroalgae, a known settlement cue for the species. We tested the hypothesis that pueruli conditioned in reduced-pH seawater would be less responsive to Laurencia spp. chemical cues than pueruli in ambient-pH seawater by comparing the proportion of individuals that moved to the cue side of the chamber with the proportion that moved to the side with no cue. We also recorded the amount of time (sec) before a response was observed. Pueruli conditioned in reduced-pH seawater were less responsive and failed to select the Laurencia cue. Our results suggest that episodic acidification of coastal waters might limit the ability of pueruli to locate settlement habitats, increasing postsettlement mortality.

Blame it on the injury: Trauma is a risk factor for pancreatic fistula following distal pancreatectomy compared with elective resection.

BACKGROUND: Postoperative pancreatic fistula (POPF) remains a significant source of morbidity following distal pancreatectomy (DP). There is a lack of information regarding the impact of trauma on POPF rates when compared with elective resection. We hypothesize that trauma will be a significant risk factor for the development of POPF following DP. METHODS: A retrospective, single-institution review of all patients undergoing DP from 1999 to 2017 was performed. Outcomes were compared between patients undergoing DP for traumatic injury to those undergoing elective resection. Univariate and multivariable analyses were performed using SAS (version 9.4). RESULTS: Of the 372 patients who underwent DP during the study period, 298 met inclusion criteria: 38 DPs for trauma (TDP), 260 elective DPs (EDP). Clinically significant grade B or C POPFs occurred in 17 (44.7%) of 38 TDPs compared with 41 (15.8%) of 260 EDPs (p < 0.0001). On multivariable analysis, traumatic injury was found to be independently predictive of developing a grade B or C POPF (odds ratio, 4.3; 95% confidence interval, 2.10-8.89). Age, sex, and wound infection were highly correlated with traumatic etiology and therefore were not retained in the multivariable model. When analyzing risk factors for each group (trauma vs. elective) separately, we found that TDP patients who developed POPFs had less sutured closure of their duct, higher infectious complications, and longer hospital stays, while EDP patients that suffered POPFs were more likely to be male, younger in age, and at a greater risk for infectious complications. Lastly, in a subgroup analysis involving only patients with drains left postoperatively, trauma was an independent predictor of any grade of fistula (A, B, or C) compared with elective DP (odds ratio, 8.6; 95% confidence interval, 3.09-24.15), suggesting that traumatic injury is risk factor for pancreatic stump closure disruption following DP. CONCLUSION: To our knowledge, this study represents the largest cohort of patients comparing pancreatic leak rates in traumatic versus elective DP, and demonstrates that traumatic injury is an independent risk factor for developing an ISGPF grade B or C pancreatic fistula following DP. LEVEL OF EVIDENCE: Prognostic study, Therapeutic, level III.

Tobacco Smoking Associated With Increased Anastomotic Disruption Following Pancreaticoduodenectomy.

BACKGROUND: The effect of cigarette smoking on postoperative morbidity following pancreaticoduodenectomy (PD) for cancer is unclear. We hypothesize that smoking is associated with higher morbidity following PD. METHODS: A retrospective review of patients undergoing PD for cancer from 2010 to 2016 at a single institution was performed. Patients who had never smoked were compared to current or past-smokers with at least 1 pack-year history. Univariate and multivariable analyses were performed. RESULTS: Two hundred fifty-two patients met inclusion criteria. On univariate analysis, there was a significant difference between smokers and never-smokers in age at diagnosis (65.5 versus 68.6 y, P = 0.013) and fistula rate (28.5% versus 16.2%, P = 0.024). Male sex was significantly associated with fistula rate compared with female sex (15.5% versus 7.1%, P = 0.023). Comparing males and females separately, smoking correlated with higher fistula development only in the male cohort (22.5% versus 5.8%, P = 0.016 in men and 7.3% versus 9.1%, P = 1.00 in women). On multivariable analysis, current and past smoking was independently predictive of developing a fistula: odds ratio of 2.038 (P = 0.030). For current and past-smokers, male sex was an independent risk factor for developing a fistula: odds ratio 2.817 (P = 0.022). There were no other significant differences between groups in rates of postoperative complications. CONCLUSIONS: Smoking status is independently predictive of postoperative pancreatic fistula following PD for cancer. Among smokers, male sex is an independent risk factor for fistula. Further studies are needed to determine if smoking cessation before surgery decreases this risk, and if so, the optimal duration of cessation.

A binomial sequential sampling plan for Bactericera cockerelli (Hemiptera: Triozidae) in Solanum lycopersicum (Solanales: Solanacea).

The tomato-potato psyllid Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) is a pest of many solanaceous plants, including tomato (Solanum lycopersicum L.) and potato (Solanum tuberosum L.). In tomato, feeding by nymphs is associated with "psyllid yellows." B. cockerelli also vectors "Candidatus Liberibacter psyllaurous," an infectious bacterium that causes "vein greening" disease. Decisions about management action are much more effective when guided by robust sampling. However, there are few previous studies of potato psyllid spatial distribution in tomato fields, and no published sequential sampling plans for the pest in tomato. We studied B. cockerelli in various tomato fields in California and used these data to generate a sequential sampling plan. We found that juvenile B. cockerelli in tomato fields exhibit an edge effect, an aggregated distribution, and individuals are primarily located on the bottom of leaves. Psyllids were concentrated in the upper segments of plants, but this changed over time. Finally, we present three binominal sequential sampling plans for managing tomato psyllids in tomato fields. These plans differed from both those for bell pepper (Capsicum annum L.) and potato, indicating that B. cockerelli needs to be sampled using crop-specific sampling plans.

Heritability and genomics of gene expression in peripheral blood.

We assessed gene expression profiles in 2,752 twins, using a classic twin design to quantify expression heritability and quantitative trait loci (eQTLs) in peripheral blood. The most highly heritable genes (∼777) were grouped into distinct expression clusters, enriched in gene-poor regions, associated with specific gene function or ontology classes, and strongly associated with disease designation. The design enabled a comparison of twin-based heritability to estimates based on dizygotic identity-by-descent sharing and distant genetic relatedness. Consideration of sampling variation suggests that previous heritability estimates have been upwardly biased. Genotyping of 2,494 twins enabled powerful identification of eQTLs, which we further examined in a replication set of 1,895 unrelated subjects. A large number of non-redundant local eQTLs (6,756) met replication criteria, whereas a relatively small number of distant eQTLs (165) met quality control and replication standards. Our results provide a new resource toward understanding the genetic control of transcription.

A sequential binomial sampling plan for potato psyllid (Hemiptera: Triozidae) on bell pepper (Capsicum annum).

BACKGROUND: Potato psyllids (Bactericera cockerelli Sulc) are a pest on solanaceous crop plants, including bell peppers. Potato psyllids vector Candidatus Liberibacter psyllaurous, but bell peppers (Capsicum annum L.) do not exhibit symptoms from infection. Potato psyllids show variation in spatial patterns and host choice with cultivar and plant species. Consequently, a study of spatial distribution and sampling plan specific to bell peppers is necessary for management of this insect pest, as those developed for other crops are unlikely to transfer among crops. RESULTS: Potato psyllids were evenly distributed on both sides of leaves but prefer the top two-thirds of pepper plants. Within fields, psyllids demonstrated an aggregated spatial distribution, but the edge effect observed in other crop plants was absent. Eggs and nymphs had similar spatial distributions that differed from adults. A series of nymph-based sampling plans were examined. Sampling plans based on an infestation of less than 41% of plants infested (5 nymphs plant(-1)) were statistically unacceptable, while little difference was found between the 41% infestation plan and 56% (20 nymphs plant(-1)) infestation plan. At 41%, an average of 11 and maximum of 49 samples would be necessary to make a treatment decision. CONCLUSION: The binomial sequential sampling plan presented here offers an important yet simple tool for managing potato psyllids in bell pepper.

Identification and impact of natural enemies of Bactericera cockerelli (Hemiptera: Triozidae) in Southern California.

Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) is a major pest of potato, (Solanum tuberosum L.), tomato (Solanum lycopersicum L.), and peppers (Capsicum spp.). The purpose of our research was to identify and determine the impact of natural enemies on B. cockerelli population dynamics. Through 2 yr of field studies (2009-2010) at four different sites and laboratory feeding tests, we identified minute pirate bug, Orius tristicolor (White) (Hemiptera: Anthocoridae); western bigeyed bug, Geocoris pallens Stål (Hemiptera:Geocoridae), and convergent lady beetle, Hippodamia convergens Guérin-Méneville (Coleoptera: Coccinellidae) as key natural enemies of B. cockerelli in southern California potatoes, tomatoes, and bell peppers. In natural enemy exclusion cage experiments in the potato crop and in American nightshade, Solanum americanum Miller, the number of B. cockerelli surviving was significantly greater in the closed cage treatments, thus confirming the affect natural enemies can have on B. cockerelli. We discuss how this information can be used in an integrated pest management program for B. cockerelli.

Spatial dispersion and binomial sequential sampling for the potato psyllid (Hemiptera: Triozidae) on potato.

BACKGROUND: The potato psyllid is a serious pest of potatoes. Sampling plans on potatoes for the potato psyllid have yet to be developed, thus the authors' objectives were (1) to determine the most efficient within-plant sampling unit, (2) to determine the spatial dispersion of potato psyllids in potato fields and (3) to develop a binomial sequential sampling plan for this pest. RESULTS: Significantly more potato plants were infested with potato psyllids on the edges of the field, and significantly more plants were infested with psyllids on the 'top' and 'middle' of the potato plant. Significantly more psyllids were also found on the undersides of leaves. The potato psyllid has an aggregated distribution in potato fields. Binomial sequential sampling plans were developed for three action thresholds representing 0.5, 1 and 5 psyllids per plant. The average sample numbers for these action thresholds were between 12 and 16 samples, depending on the action thresholds. However, based on the shape of the operating characteristic curve, the 0.5 and 1 sampling plans were more reliable than the 5 psyllids per plant plan. CONCLUSION: The binomial sequential sampling plans are useful for detecting potato psyllids at low levels of infestation, which will be useful for pest management purposes.

Effects of insecticides on behavior of adult Bactericera cockerelli (Hemiptera: Triozidae) and transmission of Candidatus Liberibacter psyllaurous.

The potato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Triozidae), is a serious pest of potatoes (Solanum tuberosum L.) that can cause yield loss by direct feeding on crop plants and by vectoring a bacterial pathogen, Candidatus Liberibacer psyllaurous. Current pest management practices rely on the use of insecticides to control the potato psyllid to lower disease incidences and increase yields. Although many studies have focused on the mortality that insecticides can cause on potato psyllid populations, little is known regarding the behavioral responses of the potato psyllid to insecticides or whether insecticides can decrease pathogen transmission. Thus, the objectives of this study were to determine the effects of insecticides on adult potato psyllid behaviors, the residual effects of insecticides on potato psyllid behaviors over time, and effects of these insecticides on Ca. L. psyllaurous transmission. Insecticides tested included imidacloprid, kaolin particle film, horticultural spray oil, abamectin, and pymetrozine. All insecticides significantly reduced probing durations and increased the amount of time adult psyllids spent off the leaflets, suggesting that these chemicals may be deterrents to feeding as well as repellents. Nonfeeding behaviors such as tasting, resting, and cleaning showed variable relationships with the different insecticide treatments over time. The insecticides imidacloprid and abamectin significantly lowered transmission of Ca. L. psyllaurous compared with untreated controls. The implications of our results for the selection of insecticides useful for an integrated pest management program for potato psyllid control are discussed.

Morphological and physiological regeneration in the auditory system of adult Mecopoda elongata (Orthoptera: Tettigoniidae).

Orthopterans are suitable model organisms for investigations of regeneration mechanisms in the auditory system. Regeneration has been described in the auditory systems of locusts (Caelifera) and of crickets (Ensifera). In this study, we comparatively investigate the neural regeneration in the auditory system in the bush cricket Mecopoda elongata. A crushing of the tympanal nerve in the foreleg of M. elongata results in a loss of auditory information transfer. Physiological recordings of the tympanal nerve suggest outgrowing fibers 5 days after crushing. An anatomical regeneration of the fibers within the central nervous system starts 10 days after crushing. The neuronal projection reaches the target area at day 20. Threshold values to low frequency airborne sound remain high after crushing, indicating a lower regeneration capability of this group of fibers. However, within the central target area the low frequency areas are also innervated. Recordings of auditory interneurons show that the regenerating fibers form new functional connections starting at day 20 after crushing.

Effects of terrestrial pollutants on insect parasitoids.

Parasitoids are important organisms in the regulation of insect herbivores in natural, urban, and agricultural ecosystems. The impact of pollutants acting on parasitoids has not been extensively reviewed. This prompted us to propose a falsifiable null hypothesis (pollutants have no effects on parasitoids) and two alternative hypotheses (pollution negatively or positively affects parasitoids) to assess in the available literature the effects of pollutants acting on parasitoids. We found 26 studies examining 39 biological systems that met our criteria for inclusion. Of these studies, 18 of the 39 biological systems (46.2%) supported the null hypothesis while 18 (46.2%) supported the first alternative hypothesis in which pollutants exhibited negative effects on parasitoids. Only a small percentage of the studies (7.6%, 3 of 39) supported the second alternative hypothesis suggesting that pollutants had positive effects on parasitoids. We provide a synthesis of the available data by pollution type, summarize trends for different pollutants, and suggest future areas of research.

Voracity and prey preference of insidious flower bug (hemiptera: anthocoridae) for immature stages of soybean aphid (hemiptera: aphididae) and soybean thrips (thysanoptera: thripidae).

The generalist predator, Orius insidiosus (Say), is an important natural enemy of the soybean aphid, Aphis glycines Matsumura. Soybean thrips, Neohydatothrips variabilis (Beach), serve as an important prey resource for O. insidiosus in soybeans, sustaining the predator's population before the arrival of the soybean aphid. Although generalist predators can forage on a broad range of prey, they may show distinct preferences for particular prey, attacking prey at levels disproportionate to their relative numbers. To assess the preference of O. insidiosus for soybean aphid and soybean thrips, attack rates of nymphal and adult O. insidiosus were measured in the laboratory. For both adults and nymphs, the number of prey attacked increased as more prey were provided. For nymphs, the total number of prey attacked increased as the predator matured. In general, the number of prey attacked by adult predators was relatively constant as the predator aged. Both O. insidiosus nymphs and adults displayed a preference for soybean thrips, by disproportionately attacking soybean thrips over soybean aphid regardless of the relative densities of the two prey. We discuss implications of this preference on O. insidiosus life history characteristics and the potential impact on O. insidiosus-prey dynamics in the field.

Effects of pollutants on bottom-up and top-down processes in insect-plant interactions.

Bottom-up (host plant quality) and top-down (natural enemies) forces both influence the fitness and population dynamics of herbivores. However, the impact of pollutants acting on these forces has not been examined, which prompted us to review the literature to test hypotheses regarding this area of research. A comprehensive literature search found 126 references which examined fitness components and population dynamics of 203 insect herbivores. One hundred and fifty-three of the 203 herbivores (75.4%) had fitness impacted due to bottom-up factors in polluted environments. In contrast, only 20 of the 203 (9.9%) had fitness significantly impacted due to top-down factors in polluted environments. The paucity of results for top-down factors impacting fitness does not necessarily mean that top-down factors are less important, but rather that fewer studies include natural enemies. We provide a synthesis of available data by pollution type and herbivore guild, and suggest future research to address this issue.