Sensitivity of different DNA extraction methods and PCR to detect resistance in patients with leprosy stratified by the bacilloscopic index

Introduction: Antimicrobial resistance in leprosy is an emerging problem, and the quantitative impact of low bacilloscopic indexes (BIs) on the sensitivity of molecular tests is unknown. We aimed to evaluate the sensitivity of gene sequencing for the detection of mutations related to antimicrobial resistance in Mycobacterium leprae in patients with low BIs using an analytical model. Methods: Patients with leprosy were included and divided into two groups depending on their BIs (≥ 2+ and < 2+). The sensitivities of the two DNA extraction methods were compared after amplifying and sequencing the repetitive element (RLEP), folP1, rpoB and gyrA in M. leprae. Results: We included 56 patients with leprosy: 35 had BIs less than 2+ (22 had negative slitskin smear [SSS] results) and 21 patients had BIs greater than or equal to 2+. The sensitivity of the amplification of the RLEP target and the gene sequencing of folP1, rpoB and gyrA was 50 to 70% lower in patients with a BI less than 2+ and was significantly reduced in patients with lower BIs for all targets (p < 0.001). One patient had a mutation in the folP1 gene, and 14 patients had mutations in the gyrA gene, but no mutations related to antimicrobial resistance were found. Conclusions: We can conclude that the sensitivity of molecular tests is directly related to the BI, but these tests can still detect up to 20% of the targets in patients with BIs < 2+. New strategies to improve the sensitivity for detecting antimicrobial resistance in leprosy patients and reasonable clinical criteria for follow-up and the introduction of alternative treatments must be developed

Photocontact allergic and phototoxic studies of chlorproethazine

Background: Neuriplege (R) cream was available as a non-prescription medication in France until its withdrawal from the market by regulatory authorities in January 2007. Its active ingredient is the phenothiazine chlorproethazine (CPE). Before its withdrawal, we investigated the photocontact allergic and phototoxic potential of Neuriplege (R) cream and CPE. Methods: An in vitro phototoxic study was performed in HaCaT keratinocytes using the neutral red dye phototoxic assay. Phototoxicity and photocontact allergy were assessed in humans by photopatch testing. Results: In vitro, a 1 h incubation of keratinocytes with CPE was approximately 13 times as toxic to the cells in the presence of ultraviolet light compared with incubation with the drug alone. Of two healthy volunteers initially photopatch tested to Neuriplege (R) cream on the arm, one developed a phototoxic reaction. These two volunteers were then photopatch tested to Neuriplege (R) and CPE on the back with seven additional healthy volunteers. Both of the initial study volunteers experienced a photocontact allergic reaction to Neuriplege (R) as is upon this re-exposure. Of the seven volunteers not previously exposed to Neuriplege (R) as is, five developed phototoxic reactions. Conclusions: This study demonstrates the strong phototoxic and photocontact allergic potential of CPE in Neuriplege (R) cream, and supports the decision of the French pharmaceutical regulatory authorities to withdraw it.</p

Oncostatin M receptor-beta mutations underlie familial primary localized cutaneous amyloidosis

Familial primary localized cutaneous amyloidosis (FPLCA) is an autosomal-dominant disorder associated with chronic skin itching and deposition of epidermal keratin filament-associated amyloid material in the dermis. FPLCA has been mapped to 5p13.1–q11.2, and by candidate gene analysis, we identified missense mutations in the OSMR gene, encoding oncostatin M-specific receptor β (OSMRβ), in three families. OSMRβ is a component of the oncostatin M (OSM) type II receptor and the interleukin (IL)-31 receptor, and cultured FPLCA keratinocytes showed reduced activation of Jak/STAT, MAPK, and PI3K/Akt pathways after OSM or IL-31 cytokine stimulation. The pathogenic amino acid substitutions are located within the extracellular fibronectin type III-like (FNIII) domains, regions critical for receptor dimerization and function. OSM and IL-31 signaling have been implicated in keratinocyte cell proliferation, differentiation, apoptosis, and inflammation, but our OSMR data in individuals with FPLCA represent the first human germline mutations in this cytokine receptor complex and provide new insight into mechanisms of skin itching

Risk for Major Bleeding in Patients Receiving Ticagrelor Compared With Aspirin After Transient Ischemic Attack or Acute Ischemic Stroke in the SOCRATES Study (Acute Stroke or Transient Ischemic Attack Treated With Aspirin or Ticagrelor and Patient Outcomes)

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