In Menière's disease, audiometry results corrected for patient age show an inherent upward-sloping configuration of the mean audiometric curve at all time points during the disease. The hearing pattern differs between unilateral and bilateral disease. The audiometric curve configuration may be an indicator of future bilateral disease.

Colored apocarotenoids accumulate at high concentrations in few plant species, where display a role in attraction of pollinators and seed dispersers. Among these apocarotenoids, crocins accumulate at high concentrations in the stigma of saffron and are responsible for the organoleptic and medicinal properties of this spice. Phytoene synthase and Orange protein are key for carotenoid biosynthesis and accumulation. We previously isolated four phytoene synthase genes from saffron with differential roles in carotenoid and apocarotenoid biosynthesis. However, the implications of Orange genes in the regulation of apocarotenoid accumulation are unknown. Here, we have identified two Orange genes from saffron, with different expression patterns. CsOr-a was mainly expressed in vegetative tissues and was induced by light and repressed by heat stress. Both CsOr-a and CsOr-b were expressed in stigmas but showed a different profile during the development of this tissue. The interactions of CsOr-a and CsOr-b were tested with all the four phytoene synthase proteins from saffron and with CsCCD2. None interactions were detected with CCD2 neither with the phytoene synthase 2, involved in apocarotenoid biosynthesis in saffron. The obtained results provide evidence of different mechanisms regulating the phytoene synthase enzymes in saffron by Orange for carotenoid and apocarotenoid accumulation in saffron.Nucleic acid purification and cDNA isolation. Total RNA was isolated from red stigmas 48 , following the manufacturer's protocols (Qiagen, Hilden, Germany). First-strand cDNAs were synthesized by RT from 2 µg of total RNA using an 18-base pair oligo dT primer and a first-strand cDNA synthesis kit (GE Healthcare Life Sciences, www.gelifesciences.com) according to manufacturer's instructions. These cDNAs were used as templates for PCR using specific primers for CsOr-a and CsOr-b (Suppl . Table 1). Thermal cycling parameters were 2 min at 95 °C, 35 cycles of 20 s at 95 °C, 20 s at 60 °C and 1 min 30 s at 72 °C, followed by a final extension of 5 min at 72 °C. The PCR products were separated in a 1% agarose gel, purified, ligated into pGEM-T (Promega, www. promega.com), and then introduced into E. coli. Scientific RepoRtS |(2020) 10:552 | https://doi.