The day that psoriasis was first induced by imiquimod (IMQ) was defined as day 0. On days −1 and 1, B6 mice were injected i.v. with 1 × 106 IL-23-induced macrophages which were treated with IL-23 for 72 h in vitro as described above.
After tumours had reached to a volume of 100∼200 mm around 14 days (defined as day 1), 200 μl of IO4-Hep, IO4-LMW-Hep or NAC-HCPS (each 10 mg ml−1 of PBS solution) was subcutaneously administered around the tumour daily for another 6 days.
The following days were successively defined as day 1, day 2, and so on.
When cells reached confluence after 10 days (>1 kΩ, defined as day −10 0) medium from the apical side of the cells was removed for ALI cultivation (defined as day 0), ALI cells were further cultivated for 21 days (defined as day 1 21) and medium on the basal side of the cells was changed every 2 days.
For adipocyte differentiation, fully confluent pre-adipose cells (defined as day 0) were treated for 2 days with induction medium (growth medium supplemented with 0.5 μg ml−1 insulin, 0.5 mm isobutylmethylxanthine (IBMX), 100 nm dexamethasone, 1 μm rosiglitazone).
Adipocyte differentiation was then induced similarly as the previously reported.18,23 Briefly, the cells (defined as day 0) were treated for 4 days with serum-free differentiation medium (DMEM/F-12 containing 33 μm biotin, 17 μm pantothenate, 10 μg ml−1 transferrin, 1 nm triiodothyronine (T3), 0.5 μg ml−1 insulin, 0.5 mm IBMX, 100 nm dexamethasone and 1 μm rosiglitazone).
Therefore, we conducted a case-control study with all passengers in whom illness developed early in the cruise (days 3 and 4) after embarkation (defined as day 1) and also with passengers who became ill later (day 5).
The day of sampling of the first histological or mycological positive specimen was defined as day 0 (D0).
Differentiation was started (defined as day 0) when the cells reached aournd 50% confluent (next day) by changing the media from MEF-CM to mesoderm induction media containing cardiac differentiation basal media (CDBM) and combined growth factors (Activin A + BMP4 + FGF2).
This day was defined as day 0 of the test for each species.
The sera of cases were gathered before (almost 5 days before), during and after the isolation of yeasts from blood culture, defined as day 0 (D0).
