Sodium chloride or plasmalyte-148 for patients presenting to emergency departments with diabetic ketoacidosis: A nested cohort study within a multicentre, cluster, crossover, randomised, controlled trial.

OBJECTIVE: To test the hypothesis that fluid resuscitation in the ED with plasmalyte-148 (PL) compared with 0.9% sodium chloride (SC) would result in a lower proportion of patients with diabetic ketoacidosis (DKA) requiring intensive care unit (ICU) admission. METHODS: We performed a prespecified nested cohort study at two hospitals within a cluster, crossover, open label, randomised, controlled trial comparing the effects of PL versus SC as fluid therapy for patients who presented to the ED with DKA. All patients presenting within a fixed recruitment period were included. The primary outcome was the proportion of patients admitted to ICU. RESULTS: Eighty-four patients were enrolled (SC n = 38, PL n = 46). The SC group had a lower median pH on admission (SC: 7.09 [interquartile range (IQR) 7.01-7.21], PL: 7.17 [IQR 6.99-7.26]). The median volume of intravenous fluids administered in ED was 2150 mL (IQR 2000-3200 mL; SC) and 2200 mL (IQR 2000-3450; PL); respectively. A higher proportion of patients in the SC group, 19 (50%), was admitted to ICU compared with PL group, 18 (39.1%); however, after adjustment for pH at presentation and diabetes type in a multivariable logistic regression model, the PL group did not have a significantly different rate of ICU admission compared with the SC group (odds ratio for ICU admission 0.73, 95% confidence interval 0.13-3.97, P = 0.71). CONCLUSION: Patients with DKA treated with PL compared with SC in the EDs had similar rates of requiring ICU admission.

Land-use impacts on benthic macroinvertebrate assemblages in pampean streams (Argentina).

In recent decades agriculture has intensified in the Argentine Pampa, and pesticide application has also increased. Livestock fields, although being progressively replaced by crops, are still commonly interspersed with crop fields. The objective of the present work is to assess the effects of land use on the benthic invertebrate assemblages of streams in the main Argentine agricultural region. Two areas were sampled during the 2011/12 growing season (November-March): Arrecifes, a homogeneous intensively cultivated area, and La Plata, a heterogeneous area of mixed livestock pasture, cropland and biological reserve. Nutrient concentrations in water were significantly higher in the streams surrounded by cropland. Measured pesticides in stream sediments were those most commonly used in crop production: chlorpyrifos, cypermethrin, lambda-cyhalothrin, endosulfan and its degradation product endosulfan sulfate. Detection frequency and pesticide concentrations were generally higher in streams surrounded by cropland than in streams surrounded by pasture or reserve. Macroinvertebrate assemblages were significantly different in streams with different land uses. Palaemonidae (Decapoda) and Caenidae (Ephemeroptera) were the taxa best represented in the reserve. Hyalellidae (Amphipoda) and Hirudinea were dominant at the streams surrounded by livestock fields. Within the streams surrounded by croplands, Oligochaeta and Hirudinea were best represented in La Plata while Chironomidae, Gastropoda and Oligochaeta were dominant at Arrecifes. Present evidence suggests that agrochemical applications contribute, in combination with other environmental variables, to the observed differences in macroinvertebrate assemblages in streams of different land use.

Climate-change influences on the response of macroinvertebrate communities to pesticide contamination in the Sacramento River, California watershed.

Limited studies have addressed how future climate-change scenarios may alter the effects of pesticides on biotic assemblages or the effects of exposures to repeated pulses of pesticide mixtures. We used reported pesticide-use data as input to a hydrological fate and transport model (Soil and Water Assessment Tool) under multiple climate-change scenarios to simulate spatiotemporal dynamics of pesticides mixtures in streams on a daily time-step in the Sacramento River watershed of California. We predicted that there will be increased pesticide application with warming across the watershed, especially in upstream areas. Using a statistical model describing the relationship between macroinvertebrate communities and pesticide dynamics, we found that compared to the baseline period of 1970-1999: (1) most climate-change scenarios predicted increased rainfall and warming across the watershed during 2070-2099; and (2) increasing pesticide contamination and increased impact on macroinvertebrates will likely occur in most areas of the watershed by 2070-2099; and (3) lower increases in effects of pesticides on macroinvertebrates were predicted for the downstream areas with intensive agriculture compared to some upstream areas with less-intensive agriculture. Future efforts on practical adaptation and mitigation strategies can be improved by awareness of altered threats of pesticide mixtures under future climate-change conditions.

Response of macroinvertebrate communities to temporal dynamics of pesticide mixtures: A case study from the Sacramento River watershed, California.

Pesticide pollution from agricultural field run-off or spray drift has been documented to impact river ecosystems worldwide. However, there is limited data on short- and long-term effects of repeated pulses of pesticide mixtures on biotic assemblages in natural systems. We used reported pesticide application data as input to a hydrological fate and transport model (Soil and Water Assessment Tool) to simulate spatiotemporal dynamics of pesticides mixtures in streams on a daily time-step. We then applied regression models to explore the relationship between macroinvertebrate communities and pesticide dynamics in the Sacramento River watershed of California during 2002-2013. We found that both maximum and average pesticide toxic units were important in determining impacts on macroinvertebrates, and that the compositions of macroinvertebrates trended toward taxa having higher resilience and resistance to pesticide exposure, based on the Species at Risk pesticide (SPEARpesticides) index. Results indicate that risk-assessment efforts can be improved by considering both short- and long-term effects of pesticide mixtures on macroinvertebrate community composition.

Land use effect on invertebrate assemblages in Pampasic streams (Buenos Aires, Argentina).

Agriculture and livestock may contribute to water quality degradation in adjacent waterbodies and produce changes in the resident invertebrate composition. The objective of the present study was to assess land use effects on the stream invertebrate assemblages in rural areas of the Argentine Pampa. The four sampling events were performed at six sites in four streams of the Pampa plain; two streams were sampled inside a biosphere reserve, and another one was surrounded by extensive livestock fields. The fourth stream was sampled at three sites; the upstream site was adjacent to agricultural plots, the following site was adjacent to an intensive livestock plot and the downstream site was adjacent to extensive breeding cattle plots. Higher pesticide concentrations were found at the site adjacent to agricultural plots and higher nutrient concentrations at the sites adjacent to agricultural and intensive breeding cattle plots. The invertebrate fauna were also different at these sites. Multivariate analysis showed a relationship between nutrient concentrations and taxonomic composition. Amphipoda (Hyalella curvispina) was the dominant group in the reserve and extensive breeding cattle sites, but was not present in the agricultural site. Also, Chironomidae were absent from the agricultural site while present at other sites. Gasteropoda (Biomphalaria peregrina), Zygoptera, and Hirudinea were dominant at the most impacted agricultural and intensive breeding cattle sites.

Insecticide concentrations in stream sediments of soy production regions of South America.

Concentrations of 17 insecticides were measured in sediments collected from 53 streams in soy production regions of South America (Argentina in 2011-2014, Paraguay and Brazil in 2013) during peak application periods. Although environmental regulations are quite different in each country, commonly used insecticides were detected at high frequencies in all regions. Maximum concentrations (and detection frequencies) for each sampling event ranged from: 1.2-7.4 ng/g dw chlorpyrifos (56-100%); 0.9-8.3 ng/g dw cypermethrin (20-100%); 0.42-16.6 ng/g dw lambda-cyhalothrin (60-100%); and, 0.49-2.1 ng/g dw endosulfan (13-100%). Other pyrethroids were detected less frequently. Banned organochlorines were most frequently detected in Brazil. In all countries, cypermethrin and/or lambda-cyhalothrin toxic units (TUs), based on Hyalella azteca LC50 bioassays, were occasionally>0.5 (indicating likely acute toxicity), while TUs for other insecticides were <0.5. All samples with total insecticide TU>1 were collected from streams with riparian buffer width<20 m. A multiple regression analysis that included five landscape and habitat predictor variables for the Brazilian streams examined indicated that buffer width was the most important predictor variable in explaining total insecticide TU values. While Brazil and Paraguay require forested stream buffers, there were no such regulations in the Argentine pampas, where buffer widths were smaller. Multiple insecticides were found in almost all stream sediment samples in intensive soy production regions, with pyrethroids most often occurring at acutely toxic concentrations, and the greatest potential for insecticide toxicity occurring in streams with minimum buffer width<20 m.

Newborn With a Perineal Lesion.

• Perineal groove is described as a wet sulcus lined with mucoumembrane extending from the posterior fourchette to the anterior anus • Perineal grooves are rarely described in literature but ar likely more common in practice. • The underlying pathogenesis and embryologic origin of perineal grooves are poorly understood. • Perineal grooves generally self-resolve by 1 year of age. • Surgical intervention is rarely indicated but can be considered for rare complications (infections, mucus drainage) or for cosmetic purposes, usually after 2 years of age. • Recognition of this mild anomaly can eliminate unnecessary (and potentially invasive) testing and procedures.

"Beat the Shock Clock": An Interprofessional Team Improves Pediatric Septic Shock Care.

Ideal care for septic shock (SS) is difficult. This interprofessional quality improvement intervention in a mid-volume pediatric emergency department aimed to reduce time to vascular access, fluid resuscitation, and antibiotics for SS. Intensive education, a care pathway, and an order set were applied. Outcome measures for patients with criteria for SS before and after intervention were compared. There were 43 patients pre-intervention (January 2009 to June 2011) and 63 post-intervention (June 2012 to June 2013). Median time to vascular access decreased from 37 minutes pre-intervention to 24 minutes post-intervention (p = 0.05). Median time to first fluid bolus decreased from 35 to 26 minutes (p = 0.08). Percentage of boluses delivered rapidly by pressure method increased from 21% to 74% (p < 0.0001). Median time to antibiotics decreased from 92 to 55 minutes (p = 0.02). In conclusion, a multimodal, interprofessional quality improvement intervention in a mid-sized pediatric emergency department improved the time to critical interventions for SS.

The p53 regulatory gene MDM2 is a direct transcriptional target of MYCN in neuroblastoma.

The MYCN oncogene is the major negative prognostic marker in neuroblastoma with important roles in both the pathogenesis and clinical behavior of this aggressive malignancy. MYC oncogenes activate both proliferative and apoptotic cellular pathways and, accordingly, inhibition of p53-mediated apoptosis is a prerequisite for MYC-driven tumorigenesis. To identify novel transcriptional targets mediating the MYCN-dependent phenotype, we screened a MYCN-amplified neuroblastoma cell line by using chromatin immunoprecipitation (ChIP) cloning. We identified the essential p53 inhibitor and protooncogene MDM2 as a putative target. MDM2 has multiple p53-independent functions modulating cell cycle and transcriptional events. Standard ChIP with MYCN antibodies established the binding of MYCN to a consensus E-box within the human MDM2 promoter. Oligonucleotide pull-down assays further established the capacity of MYCN to bind to this promoter region, confirming the ChIP results. Luciferase reporter assays confirmed the E-box-specific, MYCN-dependent regulation of the MDM2 promoter in MYCN-inducible neuroblastoma cell lines. Real-time quantitative PCR and Western blot analysis demonstrated a rapid increase in endogenous MDM2 mRNA and MDM2 protein upon induction of MYCN. Targeted inhibition of MYCN in a MYCN-amplified neuroblastoma cell line resulted in decreased MDM2 expression levels with concomitant stabilization of p53 and induction of apoptosis. Our finding that MYCN directly modulates baseline MDM2 levels suggests a mechanism contributing to the pathogenesis of neuroblastoma and other MYC-driven malignancies through inhibition of MYC-stimulated apoptosis.

Low-temperature pausing of cultivated mammalian cells.

There are currently two methods for maintaining cultured mammalian cells, continuous passage at 37 degrees C and freezing in small batches. We investigated a third approach, the "pausing" of cells for days or weeks at temperatures below 37 degrees C in a variety of cultivation vessels. High cell viability and exponential growth were observed after pausing a recombinant Chinese hamster ovary cell line (CHO-Clone 161) in a temperature range of 6-24 degrees C in microcentrifuge tubes for up to 3 weeks. After pausing in T-flasks at 4 degrees C for 9 days, adherent cultures of CHO-DG44 and human embryonic kidney (HEK293 EBNA) cells resumed exponential growth when incubated at 37 degrees C. Adherent cultures of CHO-DG44 cells paused for 2 days at 4 degrees C in T-flasks and suspension cultures of HEK293 EBNA cells paused for 3 days at either 4 degrees C or 24 degrees C in spinner flasks were efficiently transfected by the calcium phosphate-DNA coprecipitation method, yielding reporter protein levels comparable to those from nonpaused cultures. Finally, cultures of a recombinant CHO cell line (CHO-YIgG3) paused for 3 days at 4 degrees C, 12 degrees C, or 24 degrees C in bioreactors achieved the same cell mass and recombinant protein productivity levels as nonpaused cultures. The success of this approach to cell storage with rodent and human cell lines points to a general biological phenomenon which may have a wide range of applications for cultivated mammalian cells.

Effect of experimental hyperhomocysteinemia on cardiac structure and function in the rat.

Male Sprague-Dawley rats were subjected for 2 weeks to daily injections of homocysteine (Hcy), which increased plasma Hcy approximately 2-fold. Echocardiography indicated significant increases in left ventricular diastolic (13%) and systolic (31%) dimensions and decreases in posterior wall thickness (diastolic, -17%; systolic, -20%) in Hcy-treated animals. Slight changes were noted in the ejection fraction, systolic fractional shortening, and maximal aortic valvular blood flow velocity, but they were not statistically significant or were similar to those in vehicle controls. The results suggest that an initial effect of Hcy administration involves loss of myocardial structure without a direct influence on myocardial contractile function. Consistent with this conclusion, in vitro evaluation of the myocardial ring contractile response showed no significant difference in left ventricular maximal isometric force between the control (13.9 +/- 2.7 g/g tissue) and Hcy-injected (11.0 +/- 2.8 g/g tissue) animals.

Hypotension and bradycardia during caloric restriction in mice are independent of salt balance and do not require ANP receptor.

We hypothesized that caloric restriction (CR)-induced hypotension would correlate with increased sodium excretion through an atrial natriuretic peptide (ANP)-dependent mechanism. To test this hypothesis, the cardiovascular parameters of c57/Bl mice were measured with radiotelemetry while urine was collected. The 23-h mean blood pressure (BP) dropped from 108.6 +/- 1.8 to 92.7 +/- 2.4 mmHg, and 23-h heart rate dropped from 624 +/- 5 to 426 +/- 13 beats/min over 7 days of CR at 29 degrees C. Contrary to our hypothesis, urine sodium excretion decreased by 55% by day 7 of CR. Consistent with decreased sodium excretion was the drop in plasma ANP (from 82.4 +/- 4.3 to 68.0 +/- 5.8 pg/ml). To explore the possibility that CR lowers BP through an ANP receptor-dependent mechanism that is independent of its effect on sodium retention, we measured the cardiovascular parameters of mice deficient in the ANP receptor (NPR1(-/-)) or the ANP clearance receptor (NPR3(-/-)). Mean BP fell from 117.1 +/- 3.9 to 108.0 +/- 4.7 mmHg in the NPR1(-/-) mice and from 87.0 +/- 2.4 to 78.4 +/- 1.7 mmHg in the NPR3(-/-) mice during CR. These data indicate that the hypotension induced by CR does not depend on increased sodium excretion. Rather, it appears that the mouse responds to the low BP induced by CR with an increase in sodium reabsorption. Furthermore, circulating ANP levels and data from NPR1(-/-) and NPR3(-/-) mice suggest that the ANP pathway may not be involved in the cardiovascular response to CR.

Fluorescent proteins in animal cells for process development: optimization of sodium butyrate treatment as an example.

Fluorescent proteins expressed in mammalian cells can be quantified quickly and noninvasively with a standard fluorescence plate reader. We have previously exploited this quality in cell growth assessment (Hunt et al., 1999b). In this work, different CHO cell lines constitutively expressing fluorescent proteins were evaluated as model systems for process development and optimization. Our results demonstrate that the fluorescence of these cell lines quickly reveals conditions that might improve the overall productivity. Sodium butyrate, a well-known yet unpredictable enhancer of production, was chosen for this study. Due to the competing effects of sodium butyrate ("butyrate") on expression and cell number, the maximal overall productivity represents a compromise between enhancement of production and toxicity. Based on fluorescence only, it is possible to separate effects on cell number and specific production by combining microplate fluorescence measurements with data obtained by flow cytometry. This allows for rapid screening of different clones without counting cells or quantifying the recombinant protein, a highly attractive feature if the expression of green fluorescent protein (GFP) was correlated to that of a protein of interest. For all clones tested, negative effects of butyrate on proliferation were similar, while net enhancement of expression was characteristic for each clone. Therefore, it is necessary to optimize treatment for each individual clone. This work demonstrates that, based on the fluorescence of GFP-expresssing cell lines, it is possible to examine noninvasively three critical, generic parameters of butyrate treatment: butyrate concentration, exposure time, and culture phase at the time of addition.