Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae

Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of Mycoplasma infection, including cultivation, serological assays, and PCR, are time-consuming and require fully equipped stationary laboratories, which make them incompatible with testing in the resource-poor settings that are most relevant to this disease. We report a rapid, specific, and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for the detection of M. capricolum subsp. capripneumoniae. We developed the assay using a specific target sequence in M. capricolum subsp. capripneumoniae, as found in the genome sequence of the field strain ILRI181 and the type strain F38 and that was further evidenced in 10 field strains from different geographical regions. Detection limits corresponding to 5 × 10³ and 5 × 10⁴ cells/ml were obtained using genomic DNA and bacterial culture from M. capricolum subsp. capripneumoniae strain ILRI181, while no amplification was obtained from 71 related Mycoplasma isolates or from the Acholeplasma or the Pasteurella isolates, demonstrating a high degree of specificity. The assay produces a fluorescent signal within 15 to 20 min and worked well using pleural fluid obtained directly from CCPP-positive animals without prior DNA extraction. We demonstrate that the diagnosis of CCPP can be achieved, with a short sample preparation time and a simple read-out device that can be powered by a car battery, in <45 min in a simulated field setting.     

Resourcefulness,Desperation, Shame,Gratitude and Powerlessness: Common Themes Emerging from A Study of Food Bank Use in Northeast Scotland

There is growing policy maker and public concern about current trends in food bank use in Scotland. Yet little is known about the experiences of those seeking help from food banks in this country. This research aimed to address this issue by studying the use and operation of a food bank situated in a rich northeast city during January and June 2014. The study aimed to establish who was seeking help from the food bank, their reasons for doing so, and what those who did thought of, and dealt with the food they received from it. Consequently, an audit of the food bank''s client database, four months of participant observation based in the food bank, and seven face-to-face interviews with current and former food bank clients were conducted. The audit revealed that clients came from a range of socio-economic backgrounds, with men more likely to access it compared to women. Debt and social security benefit delays were cited as the main reasons for doing so. Qualitative data confirmed that sudden and unanticipated loss of income was a key driver of use. Resourcefulness in making donated food last as long as possible, keeping fuel costs low, and concern to minimise food waste were commonly described by participants. Desperation, gratitude, shame and powerlessness were also prevalent themes. Furthermore, clients were reluctant to ask for food they normally ate, as they were acutely aware that the food bank had little control over what it was able offer. Insights from this study suggest that recent UK policy proposals to address food poverty may have limited impact, without concomitant effort to address material disadvantage. Research is urgently required to determine the precise nature and extent of household level food insecurity in Scotland, and to consider monitoring for adverse physical and mental health outcomes for those affected by it.     

Grading Facial Nerve Function Following Combined Static and Mimetic Surgical Techniques

Objective To present a grading scale to assess the functional recovery of the facial nerve in patients who have undergone mimetic and static surgical techniques for facial reanimation. Study design This is a proposed new facial nerve grading system that will be demonstrated with specific case presentations. All patients underwent a variety of neural grafting, microvascular free-flap reconstruction, or surgical static procedures. Results The proposed facial nerve grading scale is one that has not been described previously in the literature and is applicable to a unique patient population. Its ease of use in this patient population will allow otolaryngologists to assess facial recovery accurately and quickly in cases where the facial nerve is not anatomically intact. Conclusion The proposed facial recovery grading scale provides an efficient means of grading facial recovery for a unique group of patients who previously could not be followed. The proposed scale is practical and easy to use in a clinical setting.     

Successful plasma exchange in a 34-year-old woman with diabetic ketoacidosis and a thrombotic microangiopathy

Thrombotic microangiopathy (TMA) associated with diabetic ketoacidosis (DKA) is a rare complication reported in the pediatric setting. We report a case of an adult patient with new-onset DM, DKA, and TMA who was treated successfully with therapeutic plasma exchange (TPE). The patient underwent five procedures and experienced quick recovery in her platelet count and a near-normalization of her LDH. Within 3 days, ADAMTS13 activity was reported at 40.7% (>66.8%). After a protracted hospital course, mostly focused on treating the patient's bilateral hemorrhagic chemosis, the patient was discharged on hospital day 30. TMA is associated with a spectrum of diseases such as TTP and sepsis but, to our knowledge, it has not been reported in the setting of DKA in an adult patient. Evidence supports that metabolic alterations associated with DKA and its treatment disrupt basal hemostatic mechanisms and promote a thrombotic state. Although ADAMTS13 activity was only moderately decreased, our patient responded rapidly to TPE, with a striking increase and stabilization of her PLT count that was durable beyond discharge. As reported recently, patients who have TMA with ADAMTS13 activity levels >10% have a range of diagnoses, presentations, and outcomes. Although the underlying microangiopathic process is incompletely understood, these patients may respond well to TPE, as was seen in this case.     

“Double-Down” Adrenal Vein Sampling Results in Patients with Apparent Bilateral Aldosterone Suppression: Utility of Repeat Sampling including Super-Selective Sampling

PurposeTo report outcomes of patients undergoing adrenal vein sampling (AVS) for primary aldosteronism with results indicating apparent bilateral adrenal suppression (ABAS), in which the adrenal aldosterone-to-cortisol ratios are decreased bilaterally (“double-down”) compared to the non-adrenal sample, and evaluate repeat AVS results.Materials and MethodsBetween 2003 and 2020, 762 patients underwent AVS. Twenty patients (2.6%; male, 12; female, 8; age 50.3 ± 9.7 years) with ABAS on initial AVS were identified. Ten underwent repeat AVS. Super-selective AVS (SS-AVS) was employed in 6 of 10 repeat AVS (60%). Outcomes after AVS were analyzed. A lateralization index (LI) >4 was considered an indication for adrenalectomy.ResultsRepeat AVS was diagnostic in 70% of patients (n = 7), with 6 of 7 lateralizing with LI >4 (median LI = 32.3; range 4.6–54.8) and 1 of 7 nearly lateralizing (LI = 3.5). All 7 patients underwent adrenalectomy. ABAS was redemonstrated in 3 patients (30%): 2 with unilateral adenomas on cross-sectional imaging underwent adrenalectomy despite ABAS results and 1 was lost to follow-up. Four of 6 patients (66%) who underwent SS-AVS were diagnosed with unilateral disease (median LI = 43.3; range 23.9–54.8), with one patient’s diagnosis reliant upon a single super-selective sample. In total, 9 patients underwent adrenalectomy after repeat AVS, all of whom had improved blood pressure control postoperatively. Ten patients did not undergo repeat AVS: 6 were lost to follow-up, 3 underwent medical management, and 1 underwent adrenalectomy.ConclusionsAVS should be repeated when “double-down” ABAS results are encountered. Super-selective sampling may provide worthwhile diagnostic data when employed during repeat AVS.