Discovery of the 1-naphthylamine biodegradation pathway reveals a broad-substrate-spectrum enzyme catalyzing 1-naphthylamine glutamylation.

1-Naphthylamine (1NA), which is harmful to human and aquatic animals, has been used widely in the manufacturing of dyes, pesticides, and rubber antioxidants. Nevertheless, little is known about its environmental behavior and no bacteria have been reported to use it as the growth substrate. Herein, we describe a pathway for 1NA degradation in the isolate Pseudomonas sp. strain JS3066, determine the structure and mechanism of the enzyme NpaA1 that catalyzes the initial reaction, and reveal how the pathway evolved. From genetic and enzymatic analysis, a five gene-cluster encoding a dioxygenase system was determined to be responsible for the initial steps in 1NA degradation through glutamylation of 1NA. The γ-glutamylated 1NA was subsequently oxidized to 1,2-dihydroxynaphthalene which was further degraded by the well-established pathway of naphthalene degradation via catechol. A glutamine synthetase-like (GS-like) enzyme (NpaA1) initiates 1NA glutamylation, and this enzyme exhibits a broad substrate selectivity toward a variety of anilines and naphthylamine derivatives. Structural analysis revealed that the aromatic residues in the 1NA entry tunnel and the V201 site in the large substrate-binding pocket significantly influence NpaA1's substrate preferences. The findings enhance understanding of degrading polycyclic aromatic amines, and will also enable the application of bioremediation at naphthylamine contaminated sites.

Prematurely terminated intron-retaining mRNAs invade axons in SFPQ null-driven neurodegeneration and are a hallmark of ALS.

Loss of SFPQ is a hallmark of motor degeneration in ALS and prevents maturation of motor neurons when occurring during embryogenesis. Here, we show that in zebrafish, developing motor neurons lacking SFPQ exhibit axon extension, branching and synaptogenesis defects, prior to degeneration. Subcellular transcriptomics reveals that loss of SFPQ in neurons produces a complex set of aberrant intron-retaining (IR) transcripts coding for neuron-specific proteins that accumulate in neurites. Some of these local IR mRNAs are prematurely terminated within the retained intron (PreT-IR). PreT-IR mRNAs undergo intronic polyadenylation, nuclear export, and localise to neurites in vitro and in vivo. We find these IR and PreT-IR mRNAs enriched in RNAseq datasets of tissue from patients with familial and sporadic ALS. This shared signature, between SFPQ-depleted neurons and ALS, functionally implicates SFPQ with the disease and suggests that neurite-centred perturbation of alternatively spliced isoforms drives the neurodegenerative process.

Pediatric cannabinoid hyperemesis: two cases.

Cannabinoid hyperemesis has recently been described in the literature. It is a syndrome characterized by severe nausea and hyperemesis in the setting of chronic marijuana abuse and, to date, has been described only in adults. We describe the syndrome in 2 pediatric patients, for whom extensive gastrointestinal workups failed to identify a clear cause and cessation of marijuana use resulted in the alleviation of their symptoms. As in most published adult cases, compulsive bathing was present in both of these cases.

Thromboelastography in Orthopaedic Trauma Acute Pelvic Fracture Resuscitation: A Descriptive Pilot Study.

OBJECTIVES: To describe the adjunctive use of thromboelastography (TEG) in directing initial blood component therapy resuscitation of patients with polytrauma with acute pelvic/acetabular fractures. DESIGN: Retrospective cohort review. SETTING: Level-2 trauma center. PATIENTS: Forty adult trauma activations with acute pelvic and/or acetabular fractures were treated with standard fracture care and TEG with adjuvant platelet mapping (TEG/PM) analysis to guide their initial 24-hour resuscitation. INTERVENTION: TEG with PM provided goal-directed hemostatic resuscitation using component blood products and an established hospital transfusion protocol. Transfusions were triggered by abnormal TEG/PM results and/or the presence of active hemorrhage, persistent hemorrhagic shock, and abnormal base deficit levels. MAIN OUTCOME MEASUREMENT: The correction of trauma-induced coagulopathy was determined by the return of a normal TEG/PM tracing. The numbers of component blood products transfused in the first 24 hours using TEG/PM were calculated. Subgroup analysis of transfusion requirements and differences between pelvic ring and acetabular fracture patterns were determined. RESULTS: More than 90% of patients received a transfusion of at least 1 blood product with 84% of transfusions occurring within 6 hours of admission. TEG/PM-guided resuscitation yielded greater volumes of platelets and packed red blood cells (PRBCs) versus fresh frozen plasma (FFP) (P = 0.018) with an average transfusion ratio of 2.5:1:2.8 (PRBC:FFP:platelet). There was a trend toward greater transfusion requirements in combined injuries versus pelvic ring or acetabular fractures (P = 0.08). CONCLUSION: TEG with PM is a valuable adjunct to guide the acute phase of resuscitation in patients with polytrauma with pelvic injuries because it allows a real-time assessment of the coagulation status. The routine use of TEG/PM may result in transfusion ratios of blood products different from those of the current empiric 1:1:1 guidelines. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Self-reported barriers to pediatric surgical care in Guatemala.

Access to pediatric surgical care is limited in low- and middle-income countries. Barriers must be identified before improvements can be made. This pilot study aimed to identify self-reported barriers to pediatric surgical care in Guatemala. We surveyed 78 families of Guatemalan children with surgical conditions who were seen at a pediatric surgical clinic in Guatemala City. Spanish translators were used to complete questionnaires regarding perceived barriers to surgical care. Surgical conditions included hernias, rectal prolapse, anorectal malformations, congenital heart defects, cryptorchidism, soft tissue masses, and vestibulourethral reflux. Average patient age was 8.2 years (range, 1 month to 17 years) with male predominance (62%). Families reported an average symptom duration of 3.7 years before clinic evaluation. Families traveled a variety of distances to obtain surgical care: 36 per cent were local (less than 10 km), 17 per cent traveled 10 to 50 km, and 47 per cent traveled greater than 50 km. Other barriers to surgery included financial (58.9%), excessive wait time in the national healthcare system (10. 2%), distrust of local surgeons (37.2%), and geographic inaccessibility to surgical care (10.2%). The majority of study patients required outpatient procedures, which could improve their quality of life. Many barriers to pediatric surgical care exist in Guatemala. Interventions to remove these obstacles may enhance access to surgery and benefit children in low- and middle-income countries.