The Utilization of an Opioid-Free Anesthetic for Pediatric Circumcision in an Ambulatory Surgery Center.

Circumcision is one of the most common urologic procedures performed at pediatric ambulatory centers. Emerging data on the short- and long-term effects of perioperative opioid administration has highlighted the importance of an opioid-free anesthetic regimen. We sought to evaluate the effectiveness of an opioid-free anesthetic in pediatric circumcision and its correlation with ambulatory surgery center efficiency. Patients, 3 years of age and younger, who underwent circumcision or circumcision revision by two surgeons pre and post introduction of an opioid-free anesthetic fast-track regimen at an outpatient surgical center were included. There were 100 patients included in this analysis, with 50 patients in each cohort. On univariate analysis, fast-tracking was associated with a decrease in median combined in-room and post-anesthesia care unit times (102.5 vs. 129.0 min, p-value < 0.001). This difference continued after multivariable analysis with an adjusted median combined in-room and post-anesthesia care unit time difference of -15.6 min (95% CI -34.2 to -12.7 min, p-value 0.018). In addition, the fast-track cohort received less intraoperative morphine equivalents without an increase in post-operative analgesic administration or change in postoperative questionnaire score. This demonstrates that opioid-free anesthesia may be used effectively in pediatric circumcision while also allowing for significant time savings for surgical centers.

Control of western corn rootworm via RNAi traits in maize: lethal and sublethal effects of Sec23 dsRNA.

BACKGROUND: RNA interference (RNAi) triggered by maize plants expressing RNA hairpins against specific western corn rootworm (WCR) transcripts have proven to be effective at controlling this pest. To provide robust crop protection, mRNA transcripts targeted by double-stranded RNA must be sensitive to knockdown and encode essential proteins. RESULTS: Using WCR adult feeding assays, we identified Sec23 as a highly lethal RNAi target. Sec23 encodes a coatomer protein, a component of the coat protein (COPII) complex that mediates ER-Golgi transport. The lethality detected in WCR adults was also observed in early instar larvae, the life stage causing most of the crop damage, suggesting that WCR adults can serve as an alternative to larvae for dsRNA screening. Surprisingly, over 85% transcript inhibition resulted in less than 40% protein knockdown, suggesting that complete protein knockdown is not necessary for Sec23 RNAi-mediated mortality. The efficacy of Sec23 dsRNA for rootworm control was confirmed in planta; T0 maize events carrying rootworm Sec23 hairpin transgenes showed high levels of root protection in greenhouse assays. A reduction in larval survival and weight were observed in the offspring of WCR females exposed to Sec23 dsRNA LC25 in diet bioassays. CONCLUSION: We describe Sec23 as RNAi target for in planta rootworm control. High mortality in exposed adult and larvae and moderate sublethal effects in the offspring of females exposed to Sec23 dsRNA LC25 , suggest the potential for field application of this RNAi trait and the need to factor in responses to sublethal exposure into insect resistance management programs. © 2019 Society of Chemical Industry.

RNAi targeting of rootworm Troponin I transcripts confers root protection in maize.

Western corn rootworm, Diabrotica virgifera virgifera, is the major agronomically important pest of maize in the US Corn Belt. To augment the repertoire of the available dsRNA-based traits that control rootworm, we explored a potentially haplolethal gene target, wings up A (wupA), which encodes Troponin I. Troponin I, a component of the Troponin-Tropomyosin complex, is an inhibitory protein involved in muscle contraction. In situ hybridization showed that feeding on wupA-targeted dsRNAs caused systemic transcript knockdown in D. v. virgifera larvae. The knockdown of wupA transcript, and by extension Troponin I protein, led to deterioration of the striated banding pattern in larval body muscle and decreased muscle integrity. Additionally, the loss of function of the circular muscles surrounding the alimentary system led to significant accumulation of food material in the hind gut, which is consistent with a loss of peristaltic motion of the alimentary canal. In this study, we demonstrate that wupA dsRNA is lethal in D. v. virgifera larvae when fed via artificial diet, with growth inhibition of up to 50% within two days of application. Further, wupA hairpins can be stably expressed and detected in maize. Maize expressing wupA hairpins exhibit robust root protection in greenhouse bioassays, with several maize transgene integration events showing root protection equivalent to commercial insecticidal protein-expressing maize.

Gene silencing in Tribolium castaneum as a tool for the targeted identification of candidate RNAi targets in crop pests.

RNAi shows potential as an agricultural technology for insect control, yet, a relatively low number of robust lethal RNAi targets have been demonstrated to control insects of agricultural interest. In the current study, a selection of lethal RNAi target genes from the iBeetle (Tribolium castaneum) screen were used to demonstrate efficacy of orthologous targets in the economically important coleopteran pests Diabrotica virgifera virgifera and Meligethes aeneus. Transcript orthologs of 50 selected genes were analyzed in D. v. virgifera diet-based RNAi bioassays; 21 of these RNAi targets showed mortality and 36 showed growth inhibition. Low dose injection- and diet-based dsRNA assays in T. castaneum and D. v. virgifera, respectively, enabled the identification of the four highly potent RNAi target genes: Rop, dre4, ncm, and RpII140. Maize was genetically engineered to express dsRNA directed against these prioritized candidate target genes. T0 plants expressing Rop, dre4, or RpII140 RNA hairpins showed protection from D. v. virgifera larval feeding damage. dsRNA targeting Rop, dre4, ncm, and RpII140 in M. aeneus also caused high levels of mortality both by injection and feeding. In summary, high throughput systems for model organisms can be successfully used to identify potent RNA targets for difficult-to-work with agricultural insect pests.

Measuring performance of night-time interfacility transport service.

The service reorganization is a part of the healthcare system reform. Some hospitals may not be able to provide all services in a 24-h basis. This evaluation was on all night-time (22 : 00 p.m. to 07 : 00 a.m.) interfacility transport by Alice Ho Miu Ling Hospital emergency department from January 2008 to December 2010, which were in-patients from nonemergency wards. The safety, speed, and performance were analyzed. During the study period, 73 cases were transferred out. Majority of them were having neurosurgical emergency (n=21, 29%) or surgical emergency (n=34, 46%). En-route physiological deteriorations were encountered in 4% (3/73) of cases. The mean acceptance time was 8 min and the team mobilization time was 13 min on average. The total service time ranged from 40 to 115 min. In conclusion, en-route adverse event was not rare. The specialized team can act as a facilitator and coordinator to improve the safety and effectiveness of the whole process.

Evaluation of a simplified therapeutic intervention scoring system (TISS-28) and the modified early warning score (MEWS) in predicting physiological deterioration during inter-facility transport.

INTRODUCTION: There is an emerging demand for inter-facility transport (IFT) of patients in recent years following changes in the healthcare framework in Hong Kong but this carries certain risks. Anticipation of possible deterioration of patients is important for patient safety and therefore risk stratification of patients before transport is important. OBJECTIVE: This study evaluated the simplified therapeutic intervention scoring system (TISS-28) and modified early warning score (MEWS) in predicting physiological deterioration en route. METHODS: This is a prospective single centre study of all emergency IFT for adult patients, excluding patients with obstetric conditions, occurring between 1 January 2005 and 30 June 2006. The severity of illness was quantified in terms of TISS-28 and MEWS. Mann-Whitney test and receiver operator characteristic (ROC) curves were used to illustrate and compare their performance. RESULTS: Among 102 patients requiring IFT, 28 had physiological deterioration en route (27%). The TISS-28 scores upon dispatch ranged from 5 to 34 with a mean of 16.5+/-5.71 whereas MEWS ranged from 0 to 11 with a mean of 2.82+/-2.01. The incidence of physiological deterioration en route was significantly greater with a higher MEWS score (P=0.001) but this was not seen with the TISS-28 score. The area under the ROC curve for the predictive value of MEWS was 0.71 which performed better than TISS-28 (area under the curve=0.53). CONCLUSION: IFT represents a group of patients with vast heterogeneity. TISS-28 is not a useful tool for risk stratification prior to transport. MEWS was able to identify patients at risk but was not ideal.

Oxidative stress after subarachnoid hemorrhage in gp91phox knockout mice.

BACKGROUND: Oxidative stress largely contributes to early brain injury after subarachnoid hemorrhage (SAH). One of the major sources of reactive oxygen species is NADPH oxidase, upregulated after SAH. We hypothesized that NADPH oxidase-induced oxidative stress plays a major causative role in early brain injury after SAH. METHODS: Using gp91phox knockout (ko) and wild-type (wt) mice, we studied early brain injury in the endovascular perforation model of SAH. Mortality rate, cerebral edema, oxidative stress, and superoxide production were measured at 24 h after SAH. Neurological evaluation was done at 23 h after SAH surgery. RESULTS: Genotyping confirmed the existence of a nonfunctional gp91phox gene in the ko mice. CBF measurements did not show differences in SAH-induced acute ischemia between ko and wt mice. SAH caused a significant increase of water content in the ipsilateral hemisphere as well as an increase of Malondialdehyde (MDA) levels and superoxide production. There were no significant differences in post-SAH mortality rate, brain water content and the intensity of the oxidative stress between knockout and wild type groups of mice. CONCLUSIONS: Our results suggest that gp91phox is not critically important to the early brain injury after SAH. An adaptive compensatory mechanism for free radical production in knockout mice is discussed.

NADPH oxidase inhibition improves neurological outcomes in surgically-induced brain injury.

Neurosurgical procedures can result in brain injury by various means including direct trauma, hemorrhage, retractor stretch, and electrocautery. This surgically-induced brain injury (SBI) can cause post-operative complications such as brain edema. By creating a mouse model of SBI, we tested whether NADPH oxidase, an important reactive oxygen species producing enzyme, is involved in SBI using transgenic mice lacking gp91phox subunit of NADPH oxidase (gp91phox KO) and apocynin, a specific inhibitor of NADPH oxidase. Neurological function and brain edema were evaluated at 24 h post-SBI in gp91phox KO and wild-type littermates grouped into SBI and sham-surgery groups. Alternatively, mice were grouped into vehicle- and apocynin-treated (5 mg/kg, i.p. 30 min before SBI) groups. Oxidative stress indicated by lipid peroxidation (LPO) was measured at 3 and 24 h post-SBI. The gp91phox KO mice, but not the apocynin-treated mice showed significantly improved neurological scores. Brain edema was observed in both gp91phox KO and wild-type groups after SBI; however, there was no significant difference between these two groups. Brain edema was also not affected by apocynin-pretreatment. LPO levels were significantly higher in SBI group in both gp91phox KO and wild-type groups as compared to sham group. A trend, although without statistical significance, was noted towards attenuation of LPO in the gp91phox KO animals as compared to wild-type group. LPO levels were significantly attenuated at 3 h post-SBI by apocynin-pretreatment but not at 24 h post-SBI. These results suggest that chronic and acute inhibition of NADPH oxidase activity does not reduce brain edema after SBI. Long-term inhibition of NADPH oxidase, however improves neurological functions after SBI.

Angiotensin I converting enzyme inhibitory peptides from in vitro pepsin-pancreatin digestion of soy protein.

Angiotensin I converting enzyme (ACE) inhibitory activity was determined in the soy protein isolate (SPI) digest produced by in vitro pepsin-pancreatin sequential digestion. The inhibitory activity was highest within the first 20 min of pepsin digestion and decreased upon subsequent digestion with pancreatin. An IC(50) value of 0.28 +/- 0.04 mg/mL was determined after 180 min of digestion, while no ACE inhibitory activity was measured for the undigested SPI at 0.73 mg/mL. Chromatographic fractionation of the SPI digest resulted in IC(50) values of active fractions ranging from 0.13 +/- 0.03 to 0.93 +/- 0.08 mg/mL. Although many of the fractions showed ACE inhibition, peptides with lower molecular masses and higher hydrophobicities were most active. The findings show that many different peptides with ACE inhibitory activities were produced after in vitro pepsin-pancreatin digestion of SPI and lead to the speculation that physiological gastrointestinal digestion could also yield ACE inhibitory peptides from SPI.

STUDY ON ANGIOTENSIN CONVERTING ENZYME INHIBITORY PEPTIDES FROM DIGESTION OF SOY PROTEIN IN VITRO / 营养学报

Objective: To investigate if angiotensin converting enzyme inhilitory(ACEI) peptides would be produced from SPI digested by a batch digestion system using enzymes similar to digestive enzymes in humans.Method: Simulate the conditions of human gastrointestinal digestion in a model digestion system in vitro and produce soy peptides from SPI digested using pepsin and pancreatin.In addition to monitoring ACEI activity in the total soy protein digest,the possibility of generating soy peptide fractions with more potent activity than the unfractionated digest was investigated by measuring activity of fractions obtained after ultrafitration,anion exchange,and RP-HPLC.Results: The generation of ACEI activity in SPI was determined after sequential digestion with pepsin and pancreatin.The inhibitory activity was highest within the first 20 min at pepsin digestion and decreased upon subsequent digestion with pancreatin.An IC50 value of 0.28?0.06 mg/ml was determined after 180 min of digestion,while no ACEI activity was measured for the undigested SPI at 0.73 mg/ml.Chromatographic fractionation of the SPI digest resulted in IC50 values of active fractions ranging from 0.13?0.03 to 0.93?0.08 mg/ml.Conclusion: Many different peptides with ACEI activities were produced after pepsin-pancreatin digestion of SPI in vitro and lead to the speculation that physiological gastrointestinal digestion could also yield ACE inhibitory peptides from SPI.