Neuromuscular Blockade Monitoring: Having It but Knowing When Not to Trust It.

Butyrylcholinesterase (BChE) is an enzyme involved in the degradation of depolarizing and non-depolarizing neuromuscular blocking agents (NMBA), such as succinylcholine and mivacurium, respectively. Its deficiency is inherited or acquired, and results in paralysis of skeletal muscles after NMBA administration. We report a case of a 32-year-old pregnant woman proposed for cesarean section. General anesthesia (GA) was induced using propofol and succinylcholine. The surgical procedure was uneventful but after 40 minutes, there was no reversal of neuromuscular block (NMB). Other differential diagnoses were excluded and a deficit of BChe was assumed. When the train-of-four ratio (TOFr) achieved 40%, neostigmine/atropine led to the slow recovery of NMB up to TOFr 88%. The patient was extubated, but ventilation proved ineffective, so GA was induced and the patient was reintubated. A new measurement found a TOFr of 60%. Sedation and ventilatory support were maintained until the complete reversal of NMB (4 hours after succinylcholine). Prolonged block is a rare but serious complication of the use of succinylcholine in patients with BChE deficiency. This report not only highlights the importance of intraoperative NMB monitoring in homozygotic patients for atypical cholinesterase but also raises awareness for its careful interpretation.

Inadequate Adenosine-Induced Flow Arrest During Intraoperative Basilar Aneurysm Surgery.

Aneurysmal subarachnoid hemorrhage (SAH) is an acute neurologic emergency. We report the case of a 48-year-old male with a massive SAH caused by a ruptured aneurysm of the vertebrobasilar transition. During an urgent craniotomy, due to an aneurysm re-rupture, adenosine was given for flow arrest but no sinus pause was observed. Esmolol was administered and strategies for cerebral protection were implemented. The surgeon was able to clip the aneurysm and the patient was discharged after 78 days without sequelae. The highest adenosine dose given did not result in an efficient cardiac pause. Atropine given one hour before could have contributed to this. This case highlights a successfully managed case of ruptured aneurysm with refractory adenosine-induced flow arrest.

Disabled insecticidal proteins: A novel tool to understand differences in insect receptor utilization.

The development of insect resistance to pesticides via natural selection is an acknowledged agricultural issue. Likewise, resistance development in target insect populations is a significant challenge to the durability of crop traits conferring insect protection and has driven the need for novel insecticidal proteins (IPs) with alternative mechanism of action (MOA) mediated by different insect receptors. The combination or "stacking" of transgenes encoding different insecticidal proteins in a single crop plant can greatly delay the development of insect resistance, but requires sufficient knowledge of MOA to identify proteins with different receptor preferences. Accordingly, a rapid technique for differentiating the receptor binding preferences of insecticidal proteins is a critical need. This article introduces the Disabled Insecticidal Protein (DIP) method as applied to the well-known family of three-domain insecticidal proteins from Bacillus thuringiensis and related bacteria. These DIP's contain amino acid substitutions in domain 1 that render the proteins non-toxic but still capable of competing with active proteins in insect feeding assays, resulting in a suppression of the expected insecticidal activity. A set of insecticidal proteins with known differences in receptor binding (Cry1Ab3, Cry1Ac.107, Cry2Ab2, Cry1Ca, Cry1A.105, and Cry1A.1088) has been studied using the DIP method, yielding results that are consistent with previous MOA studies. When a native IP and an excess of DIP are co-administered to insects in a feeding assay, the outcome depends on the overlap between their MOAs: if receptors are shared, then the DIP saturates the receptors to which the native protein would ordinarily bind, and acts as an antidote whereas, if there is no shared receptor, the toxicity of the native insecticidal protein is not inhibited. These results suggest that the DIP methodology, employing standard insect feeding assays, is a robust and effective method for rapid MOA differentiation among insecticidal proteins.

Use of exome sequencing to determine the full profile of genetic variants in the fluoropyrimidine pathway in colorectal cancer patients affected by severe toxicity.

AIM: To identify genetic variants associated with capecitabine toxicity in fluoropyrimidine pathway genes using exome sequencing. PATIENTS & METHODS: Exomes from eight capecitabine-treated patients with severe adverse reactions (grade >2), among a population of 319, were sequenced (Ion Proton). SNPs in genes classified as potentially damaging (Sorting Intolerant from Tolerant and Polymorphism Phenotyping v2) were tested for association with toxicity in a validation cohort of 319 capecitabine-treated patients. RESULTS: A total of 17 nonsynonymous genetic variants were identified. Of these, five putative damaging SNPs in DPYD, ABCC4 and MTHFR were genotyped in the validation cohort. DPYD rs1801160 was associated with the risk of toxicity (p = 0.029) and MTHFR rs1801133 with delayed administration of chemotherapy due to toxicity (p = 0.047). CONCLUSION: Exome sequencing revealed two specific biomarkers of the risk of toxicity to capecitabine.

Greenhouse gas balance of mountain dairy farms as affected by grassland carbon sequestration.

Recent studies on milk production have often focused on environmental impacts analysed using the Life Cycle Assessment (LCA) approach. In grassland-based livestock systems, soil carbon sequestration might be a potential sink to mitigate greenhouse gas (GHG) balance. Nevertheless, there is no commonly shared methodology. In this work, the GHG emissions of small-scale mountain dairy farms were assessed using the LCA approach. Two functional units, kg of Fat and Protein Corrected Milk (FPCM) and Utilizable Agricultural Land (UAL), and two different emissions allocations methods, no allocation and physical allocation, which accounts for the co-product beef, were considered. Two groups of small-scale dairy farms were identified based on the Livestock Units (LU) reared: <30 LU (LLU) and >30 LU (HLU). Before considering soil carbon sequestration in LCA, performing no allocation methods, LLU farms tended to have higher GHG emission than HLU farms per kg of FPCM (1.94 vs. 1.59 kg CO2-eq/kg FPCM, P ≤ 0.10), whereas the situation was reversed upon considering the m2 of UAL as a functional unit (0.29 vs. 0.89 kg CO2-eq/m2, P ≤ 0.05). Conversely, considering physical allocation, the difference between the two groups became less noticeable. When the contribution from soil carbon sequestration was included in the LCA and no allocation method was performed, LLU farms registered higher values of GHG emission per kg of FPCM than HLU farms (1.38 vs. 1.10 kg CO2-eq/kg FPCM, P ≤ 0.05), and the situation was likewise reversed in this case upon considering the m2 of UAL as a functional unit (0.22 vs. 0.73 kg CO2-eq/m2, P ≤ 0.05). To highlight how the presence of grasslands is crucial for the carbon footprint of small-scale farms, this study also applied a simulation for increasing the forage self-sufficiency of farms to 100%. In this case, an average reduction of GHG emission per kg of FPCM of farms was estimated both with no allocation and with physical allocation, reaching 27.0% and 28.8%, respectively.

Evaluating seasonal dynamics of bacterial communities in marine fish aquaculture: a preliminary study before applying phage therapy.

The increasing problem of antibiotic resistance in common pathogenic bacteria and the concern about the spreading of antibiotics in the environment bring the need to find new methods to control fish pathogens. Phage therapy represents a potential alternative to antibiotics, but its use in aquaculture requires a detailed understanding of bacterial communities, namely of fish pathogenic bacteria. Therefore, in this study the seasonal dynamics of the overall bacterial communities, microbiological water quality and disease-causing bacteria were followed in a marine aquaculture system of Ria de Aveiro (Portugal). Analysis of the bacterial diversity of the water samples by denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments indicates that the bacterial community structure varied seasonally, showing a higher complexity during the warm season. The diversity of the main fish pathogenic bacteria, assessed by DGGE targeting the Vibrio genus, showed lower seasonal variation, with new dominating populations appearing mainly in the spring. Bacterial indicators, faecal coliforms and enterococci, enumerated by the filter-membrane method, also varied seasonally. The fluorescent in situ hybridization (FISH) results showed that the specific groups of bacteria varied during the study period and that the non-indigenous Enterobactereaceae family was the most abundant group followed by Vibrio and Aeromonas. The seasonal variation detected in terms of density and structure of total and pathogenic bacterial communities demonstrates the need for a careful monitoring of water through the year in order to select the suitable phages to inactivate fish pathogenic bacteria. The spring season seems to be the critical time period when phage therapy should be applied.