An Optimized Small-Scale Rearing System to Support Embryonic Microinjection Protocols for Western Corn Rootworm, Diabrotica virgifera virgifera.

Western corn rootworm (WCR), a major pest of corn, has been reared in laboratories since the 1960s. While established rearing methods are appropriate for maintaining WCR colonies, they are not optimal for performing germline transformation or CRISPR/Cas9-based genome editing. Here we report the development of an optimized rearing system for use in WCR functional genomics research, specifically the development of a system that facilitates the collection of preblastoderm embryos for microinjection as well as gathering large larvae and pupae for downstream phenotypic screening. Further, transgenic-based experiments require stable and well-defined survival rates and the ability to manipulate insects at every life stage. In our system, the WCR life cycle (egg to adult) takes approximately 42 days, with most individuals eclosing between 41 and 45 days post oviposition. Over the course of one year, our overall survival rate was 67%. We used this data to establish a quality control system for more accurately monitoring colony health. Herein, we also offer detailed descriptions for setting up single-pair crosses and conducting phenotypic screens to identify transgenic progeny. This study provides a model for the development of new rearing systems and the establishment of highly controlled processes for specialized purposes.

How Rearing Systems for Various Species of Flies Benefit Humanity.

Flies (Diptera) have played a prominent role in human history, and several fly species are reared at different scales and for different beneficial purposes worldwide. Here, we review the historical importance of fly rearing as a foundation for insect rearing science and technology and synthesize information on the uses and rearing diets of more than 50 fly species in the families Asilidae, Calliphoridae, Coelopidae, Drosophilidae, Ephydridae, Muscidae, Sarcophagidae, Stratiomyidae, Syrphidae, Tachinidae, Tephritidae, and Tipulidae. We report more than 10 uses and applications of reared flies to the well-being and progress of humanity. We focus on the fields of animal feed and human food products, pest control and pollination services, medical wound therapy treatments, criminal investigations, and on the development of several branches of biology using flies as model organisms. We highlight the relevance of laboratory-reared Drosophila melanogaster Meigen as a vehicle of great scientific discoveries that have shaped our understanding of many biological systems, including the genetic basis of heredity and of terrible diseases such as cancer. We point out key areas of fly-rearing research such as nutrition, physiology, anatomy/morphology, genetics, genetic pest management, cryopreservation, and ecology. We conclude that fly rearing is an activity with great benefits for human well-being and should be promoted for future advancement in diverse and innovative methods of improving existing and emerging problems to humanity.

COVID-19 Vaccine Provider Access and Vaccination Coverage Among Children Aged 5-11 Years - United States, November 2021-January 2022.

On October 29, 2021, the Pfizer-BioNTech pediatric COVID-19 vaccine received Emergency Use Authorization for children aged 5-11 years in the United States.† For a successful immunization program, both access to and uptake of the vaccine are needed. Fifteen million doses were initially made available to pediatric providers to ensure the broadest possible access for the estimated 28 million eligible children aged 5-11 years, especially those in high social vulnerability index (SVI)§ communities. Initial supply was strategically distributed to maximize vaccination opportunities for U.S. children aged 5-11 years. COVID-19 vaccination coverage among persons aged 12-17 years has lagged (1), and vaccine confidence has been identified as a concern among parents and caregivers (2). Therefore, COVID-19 provider access and early vaccination coverage among children aged 5-11 years in high and low SVI communities were examined during November 1, 2021-January 18, 2022. As of November 29, 2021 (4 weeks after program launch), 38,732 providers were enrolled, and 92% of U.S. children aged 5-11 years lived within 5 miles of an active provider. As of January 18, 2022 (11 weeks after program launch), 39,786 providers had administered 13.3 million doses. First dose coverage at 4 weeks after launch was 15.0% (10.5% and 17.5% in high and low SVI areas, respectively; rate ratio [RR] = 0.68; 95% CI = 0.60-0.78), and at 11 weeks was 27.7% (21.2% and 29.0% in high and low SVI areas, respectively; RR = 0.76; 95% CI = 0.68-0.84). Overall series completion at 11 weeks after launch was 19.1% (13.7% and 21.7% in high and low SVI areas, respectively; RR = 0.67; 95% CI = 0.58-0.77). Pharmacies administered 46.4% of doses to this age group, including 48.7% of doses in high SVI areas and 44.4% in low SVI areas. Although COVID-19 vaccination coverage rates were low, particularly in high SVI areas, first dose coverage improved over time. Additional outreach is critical, especially in high SVI areas, to improve vaccine confidence and increase coverage rates among children aged 5-11 years.

Optimization of a Diet for the Greater Wax Moth (Lepidoptera: Pyralidae) Using Full Factorial and Mixture Design.

Diet optimization is an important process to increase the efficiency of rearing insects and can be used to develop high-quality insects with specific fitness and life-history traits. Galleria mellonella (L.), the greater wax moth, is widely used in research, microbiology assays, as pet food, and host for biological control agents. Although artificial diets for G. mellonella have been researched and optimized for decades, preliminary tests indicated that the predominantly utilized G. mellonella diet could be improved to yield larger larvae with a short development time. We used a design of experiments (DOE) approach that incorporated multiple full factorial designs and a final mixture design to test the qualitative and quantitative effects of ingredients and their interactions on larval mass and survival. Analysis of 17 ingredient variations in 35 diet formulations yielded an optimized diet that supported high survival and 2.4-fold greater larval body mass than the standard rearing diet. This study demonstrates the importance and efficiency of statistical DOE in guiding the optimization of insect diets to improve traits that represent the quality and fitness of the reared insects.

Varroa destructor feeds primarily on honey bee fat body tissue and not hemolymph.

The parasitic mite Varroa destructor is the greatest single driver of the global honey bee health decline. Better understanding of the association of this parasite and its host is critical to developing sustainable management practices. Our work shows that this parasite is not consuming hemolymph, as has been the accepted view, but damages host bees by consuming fat body, a tissue roughly analogous to the mammalian liver. Both hemolymph and fat body in honey bees were marked with fluorescent biostains. The fluorescence profile in the guts of mites allowed to feed on these bees was very different from that of the hemolymph of the host bee but consistently matched the fluorescence profile unique to the fat body. Via transmission electron microscopy, we observed externally digested fat body tissue in the wounds of parasitized bees. Mites in their reproductive phase were then fed a diet composed of one or both tissues. Mites fed hemolymph showed fitness metrics no different from the starved control. Mites fed fat body survived longer and produced more eggs than those fed hemolymph, suggesting that fat body is integral to their diet when feeding on brood as well. Collectively, these findings strongly suggest that Varroa are exploiting the fat body as their primary source of sustenance: a tissue integral to proper immune function, pesticide detoxification, overwinter survival, and several other essential processes in healthy bees. These findings underscore a need to revisit our understanding of this parasite and its impacts, both direct and indirect, on honey bee health.

Prevalence of hiatal hernia in the morbidly obese.

BACKGROUND: Morbidly obese patients commonly have gastroesophageal reflux (GERD) and associated hiatal hernias. As such, some surgeons routinely perform a concomitant hiatal hernia repair during bariatric surgery. However, the intraoperative inspection for a hiatal hernia based on laparoscopic visualization can be misleading. The aim of this study was to assess the prevalence of hiatal hernias in morbidly obese patients based on preoperative upper gastrointestinal (GI) contrast study. METHODS: Data on 181 patients who underwent routine upper GI contrast study as part of a preoperative workup for bariatric surgery were reviewed. The upper GI studies were examined for the presence of hiatal hernias and GERD. Hiatal hernias were categorized by size as small (≤2 cm), moderate (2-5 cm), or large (>5 cm). GERD was based on radiologic evidence and categorized as mild, moderate, or severe. RESULTS: The mean age of the cohort was 44 years, with a mean body mass index of 43 kg/m(2). Of the 181 patients overall, based on the upper GI contrast study, the prevalence of hiatal hernia was 37.0% and of GERD was 39.8%; the prevalence of moderate or large hiatal hernia was 4.4%, and the prevalence of moderate or severe GERD was 13.3%. CONCLUSIONS: Based on upper GI contrast study, we identified the presence of a hiatal hernia in nearly 40% of morbidly obese patients. The results from this study suggest that surgeons should evaluate the morbidly obese patient for the presence of hiatal hernias and perform concomitant repair at the time of the bariatric procedure, particularly in patients undergoing gastric banding and sleeve gastrectomy, while less so in the gastric bypass patient.

Vision rehabilitation for visual-vestibular dysfunction: the role of the neuro-optometrist.

INTRODUCTION: This article discusses, in a clinically relevant format, the importance of including a neuro-optometrist as a member of the management team for patients with balance disorders. PURPOSE: To review the importance of vision and visual processing for maintaining a sense of balance and equilibrium and the role of the neuro-optometrist in the overall rehabilitation of patients with balance disorders SUMMARY: Dizziness, balance problems and the sensation that the space world is moving (vertigo) are one of the most commonly reported problems in general medical practice. Persons with a central nervous system injury or other idiopathic causes of visual processing problems or who have functional vision problems that are not adequately managed, often experience extreme difficulty with balance and movement, as well as with their perception of space. Consequently, the patient often experiences difficulty functioning in an environment with excessive visual stimulation such as a grocery store or shopping mall. Symptoms of disequilibrium, vestibular and balance problems are commonly a result of VOR disturbance secondary to an inner ear problem and an unstable binocularity. CONCLUSION: The combination of neuro-optomertic rehabilitative therapy and balance therapy will result in a is an effective treatment for reducing or resolving these symptoms.

Low-dose and standard computed tomography scans yield equivalent stone measurements.

OBJECTIVE: To ascertain the reliability of low-dose computed tomography (CT) compared with standard CT in the determination of stone size, density, and skin-to-stone distance (SSD). MATERIALS AND METHODS: A total of 10 patients seen in the emergency room within a mean of 23 days (range 0-51) underwent both conventional CT and low-dose CT for the same stone. The radiation dose reduction was calculated according to the patient's body mass index. The CT scans were performed with 2-mm section cuts, and 3-dimensional reconstruction was performed to obtain the coronal views. The stone size was measured (ie, height, width, and length), and the Hounsfield units were calculated. In addition, the SSD was calculated for the nonmoving renal stones. RESULTS: No difference was found in stone size between the 2 dosage levels, as measured by the height, width, length, and volume of the stone (P = .9, P = .7, P = .8, and P = .8 respectively). In addition, no difference in Hounsfield units was appreciated between the 2 scan types (P = .6). Finally, no significant difference was found in the SSD (P = .5). Between the 2 scans, the average effective dose reduction was 73%, from 23 to 6 mSv (P = .002). CONCLUSION: No significant difference was found in the measurement of stone size, Hounsfield units, or SSD between the low-dose and conventional-dose CT scans. However, the low-dose CT scans resulted in a marked reduction in the radiation dose to the patient.

CT evaluation of placental abruption in pregnant trauma patients.

The purpose of the study was to assess the possibility of placental injury detection on computed tomography (CT) in pregnant trauma patients. The images and dictated reports of 44 CT scans of pregnant women who presented to the University of California Irvine Medical Center (UCIMC) from 2003 to 2008 for traumatic abdominal conditions were reviewed for placental abruption. Performances of original dictated reports, an untrained reviewer, and a trained reviewer (who was trained on 22 non-traumatic scans) were compared. Of the 66 pregnant women who received abdominal CT scans, 44 sustained abdominal trauma. Seven suffered placental abruptions, all of which were identified on CT. Sensitivity and specificity were 100% and 79.5%, respectively, for the untrained reviewer, 100% and 82.1% for the trained reviewer, and 42.9% and 89.7% for the original dictated reports. Placental abruptions are often overlooked on CT scan. Sensitivity may be improved by systematic evaluation of the placenta and specificity by training on normal placental morphology.

Retrocecal appendix location and perforation at presentation.

Retrocecal appendicitis has been theorized to follow a more insidious course than other anatomic variants. To determine the influence of retrocecal anatomy on clinical course of appendicitis, 200 adult patients treated at a major university medical center with the diagnosis of appendicitis from 2001 to 2004 were retrospectively studied. Computed tomography (CT) scans of adult patients with an ultimate diagnosis of appendicitis were analyzed to determine an association between retrocecal appendix and perforation of the appendix at presentation. A higher perforation rate in the retrocecal group would imply patient delay in presentation from more tolerable symptoms. CT scans were examined for retrocecal location and perforation. No significant association was found between retrocecal anatomy and perforation rates at presentation (chi-square = 2.1, P = 0.15, odds ratio = 1.6, 95% confidence interval [0.8-3.0]). However, the risk of perforation was 60 per cent higher in the retrocecal group. By regression analysis, age and the presence of a fecalith on CT scan were predictors of appendix perforation. Appendix location was not. In this study, we found no significant association between retrocecal appendix anatomy and perforation at presentation.

Accurate diagnosis of infarction of omentum and appendices epiploicae by computed tomography.

Segmental infarction of the omentum and epiploic appendages presents with acute abdominal findings that may be confused with a surgical illness. Computed tomography, however, demonstrates a consistent and well-recognized pattern that allows safe, nonoperative treatment. Infarction of omental or mesenteric fat may present clinically as localized peritonitis, mimicking appendicitis, diverticulitis, or cholecystitis. Spontaneous recovery without operation is to be expected if an accurate diagnosis is established. We describe the diagnosis, treatment, and outcome of 15 patients who had infarction of the greater omentum (eight) and epiploic appendage (seven) and presented with localized abdominal pain and tenderness, with six demonstrating regional peritonitis and fever. All underwent CT imaging during their initial evaluation, and 12 of the 15 patients were diagnosed with focal omental or mesenteric fat infarction radiographically and managed nonoperatively. Three patients who had characteristic CT findings nevertheless underwent operation. All patients had complete resolution of their abdominal pain regardless of treatment. The clinical presentation of infarction of the omental or epiploic appendages may be difficult to differentiate from surgical causes of acute abdominal pain. The characteristic findings on computed tomography are diagnostic and allow safe, conservative management in the majority of patients.

Superior mesenteric artery syndrome involving the duodenum and jejunum.

Superior mesenteric artery (SMA) syndrome is a rare cause of intestinal obstruction involving the duodenum. Diagnosis is based on clinical suspicion with radiologic confirmation. We report an unusual presentation of the SMA syndrome involving both the duodenum and jejunum initially not recognized on contrast-enhanced CT. This case demonstrates the judicious use of multiple modalities in evaluating for this syndrome.

Optimizing the patient positioning for PICC line tip determination.

Central venous catheters (CVCs) are used for both emergent and long-term vascular access for the infusion of numerous therapeutic agents such as chemotherapy, parenteral nutrition, antibiotics, and analgesics, as well as for temporary hemodialysis or hemoperfusion. Current standard of care dictates that CVC insertion should be followed by an immediate chest radiograph to confirm appropriate position. Radiographic confirmation of central venous line placement is important because it is not possible to determine CVC tip position clinically. Although many catheter tips can be localized on the standard frontal radiograph, there are occasions when a second radiograph is necessary to localize the position of the CVC tip accurately. We hypothesized that a right posterior obligue chest radiograph would more consistently enable the catheter tip to be seen as it reduces the superimposition of mediastinal structures. One hundred chest radiographs taken in an anteroposterior (AP) projection and 100 chest radiographs taken in a right posterior oblique (RPO) projection after a peripherally inserted central catheter (PICC) line placement at UCI Medical Center from June 2000 to November 2002 were read by two radiologists. Forty-one percent of AP readings were discrepant and 4% had the annotation "difficult to identify the position of the tip" although the identification of tip position was similar. Fifty-five percent of AP readings were in agreement with no note of any difficulty. Eighteen percent of RPO readings were discrepant and 2% had the annotation "difficult to identify the position of the tip" although the identification of tip position was similar. Eighty percent of RPO readings were in agreement with no note of any difficulty.

Deliberately added and "cryptic" antioxidants in three artificial diets for insects.

Characteristics of both deliberately added and "cryptic" antioxidants were assayed from hydrophilic and lipophilic extracts from artificial diets for plant bugs, lepidopteran larvae, and green lacewings. Cryptic antioxidants are defined as substances naturally existing in diet ingredients but not deliberately added because of their antioxidant potential. Diets were tested after 1) being freshly produced, 2) stored for 48 h at 4 degrees C, or 3) held for 48 h under rearing room conditions at 27 degrees C. Tests included 1) a general assay of antioxidant capacity known as the ferric-reducing antioxidant power (FRAP) assay. 2) a cation radical-scavenging assay, 3) an ascorbic acid assay, and 4) an assay of inhibition of lipid peroxidation. In all assays, the lepidopteran diet had the highest values for protection against reactive oxygen species (ROS). The lepidopteran diet (with 0.17-0.23-mg equivalents of gallic acid equals total phenolic compounds per gram of diet) had three- to four-fold higher concentrations of phenolic compounds than did either the plant bug diet or the lacewing diet. Unexpectedly, the plant bug and the lacewing diets caused more lipid peroxidation than did the positive controls. This was attributed to the high concentrations of iron in these diets (mainly from chicken eggs), causing an ascorbate-ferric ion-induced lipid peroxidation. Diet storage, measured after 2 d at 27 or 4-6 degrees C, caused no significant declines in overall antioxidant potential. However, storage did lead to decline in ascorbic acid. The FRAP assay offered the best potential as a general, routine test of the potential of various insect diets to resist the destructive effects of ROS. The importance of addressing issues of protection against ROS in insect diets is discussed.

Management of traumatic abdominal wall hernia.

Traumatic abdominal wall hernia (TAWH) can occur after blunt trauma and can be classified into low- or high-energy injuries. Low energy injuries occur after impact on a small blunt object. High-energy injuries are sustained during motor vehicle accidents or automobile versus pedestrian accidents. We present six cases of high-energy TAWH cases that were treated at our trauma center. All patients presented with varying degrees of abdominal tenderness with either abdominal skin ecchymosis or abrasions, which made physical examination difficult. CT scan confirmed the hernia in each patient. All six patients had associated injuries that required open repair. The abdominal wall defects were repaired primarily. Three patients (50%) in our series developed a postoperative wound infection or abscess. Review of the literature on low-energy TAWH shows no associated abdominal injuries. In conclusion distinction between low- and high-energy injury is imperative in the management of TAWH. Hernias following low-energy injuries can be repaired after local exploration through an incision overlying the defect. TAWHs following high-energy trauma should undergo exploratory laparotomy through a midline incision. The defect should be repaired primarily and prosthetics avoided because of the high incidence of postoperative infection.

Iatrogenic venous air embolism during contrast enhanced computed tomography: a report of two cases.

Venous air embolism (VAE) is a known complication of venous access procedures such as contrast-enhanced computed tomography (CECT). Although a massive VAE can be fatal, most iatrogenic VAE cases during CECT involve a few milliliters of air and are asymptomatic. We report two cases of massive and nonfatal VAE during CECT. Both cases involve the inadvertent injection of air instead of contrast by power injectors during the contrast phase. In both cases, the patients were stable and survived the event without permanent sequelae. We also discuss the pathophysiology, treatment, and prevention of VAE, especially during CECT.

Insect feeding mobilizes a unique plant defense protease that disrupts the peritrophic matrix of caterpillars.

Plants frequently respond to herbivorous insect attack by synthesizing defense proteins that deter insect feeding and prevent additional herbivory. Maize (Zea mays L.) lines, resistant to feeding by a number of lepidopteran species, rapidly mobilize a unique 33-kDa cysteine protease in response to caterpillar feeding. The accumulation of the 33-kDa cysteine protease in the maize mid-whorl was correlated with a significant reduction in caterpillar growth that resulted from impaired nutrient utilization. Black Mexican Sweetcorn callus transformed with mir1, the gene encoding the 33-kDa cysteine protease, expressed the protease and growth of caterpillars reared on the transgenic callus was reduced 60-80%. Scanning electron microscopy was used to examine the effect of plant material expressing the 33-kDa cysteine protease on the structure of the caterpillar peritrophic matrix. Because the peritrophic matrix surrounds the food bolus, assists in digestive processes, and protects the caterpillar midgut from physical and chemical damage, disruption of peritrophic matrix may reduce caterpillar growth. The results indicated that the peritrophic matrix was severely damaged when caterpillars fed on resistant maize plants or transgenic Black Mexican Sweetcorn. The accumulation of the 33-kDa cysteine protease in response to caterpillar feeding, and its ability to damage the insect peritrophic matrix, represents an unusual host-plant resistance mechanism that may have applications in agricultural biotechnology.

Effect of antifungal agents on biological fitness of Lygus hesperus (Heteroptera: Miridae).

Artificial diets have become important components of rearing systems for insects that are used for research purposes and in commercial production. Because the rearing conditions for insects also provide ideal settings for mold growth, antifungal additives are often used to reduce diet contamination. However, the antifungal agents must not only be effective in mold suppression, they must also be safe to the target insects of the rearing programs. The toxicity of five commonly used antifungal agents (benzoic acid, formalin, methyl paraben, propionic acid, and sorbic acid) was tested using diet bioassays on Lygus hesperus Knight, and the effect on biological fitness was measured. Biological fitness was defined as total number of survivors, mean biomass (dry weight) accumulated per cage over the total treatment period, egg production, time to adult emergence, and time to start of egg laying. Methyl paraben and formalin were found to have significant negative effects on these measurements of biological fitness. Challenge tests to determine the ability of the antifungal agents to suppress mold growth when inoculated into the diet medium are currently in progress.

Molecular cloning and partial characterization of a trypsin-like protein in salivary glands of Lygus hesperus (Hemiptera: Miridae).

Trypsin-like enzymes from the salivary gland complex (SGC) of Lygus hesperus Knight were partially purified by preparative isoelectric focusing (IEF). Enzyme active against Nalpha-benzoyl-L-arginine-p-nitroanilide (BApNA) focused at approximately pH 10 during IEF. This alkaline fraction gave a single activity band when analyzed with casein zymograms. The serine proteinase inhibitors, phenylmethylsulfonyl fluoride (PMSF) and lima bean trypsin inhibitor, completely inhibited or suppressed the caseinolytic activity in the crude salivary gland extract as well as the IEF-purified sample. Chicken egg white trypsin inhibitor also inhibited the IEF-purified sample but was not effective against a major caseinolytic band in the crude salivary gland extract. These data indicated the presence of serine proteinases in the SGC of L. hesperus. Cloning and sequencing of a trypsin-like precursor cDNA provided additional direct evidence for serine proteinases in L. hesperus. The encoded trypsin-like protein included amino acid sequence motifs, which are conserved with five homologous serine proteinases from other insects. Typical features of the putative trypsin-like protein from L. hesperus included residues in the serine proteinase active site (His(89), Asp(139), Ser(229)), conserved cysteine residues for disulfide bridges, residues (Asp(223), Gly(252), Gly(262)) that determine trypsin specificity, and both zymogen signal and activation peptides.