Comparison of flow cytometry and immunohistochemistry in non-radioisotopic murine lymph node assay using bromodeoxyuridine

Hydrogen peroxide and monoethanolamine are the key causative ingredients for hair dye-induced dermatitis and hair loss

“…PPD, one of the main components of commercial oxidative hair dye products, is well-known as the potent contact sensitizer and skin irritant [8,15,16,35]. It is reported that people who is allergic to hair dye, can show hair loss, especially, telogen effluvium, a form of non-scarring alopecia characterized by diffuse hair shedding [36].…”

“…PPD, which is contained up to 6% in hair dyes as a black-coloring agent [15], is a well-known skin sensitizer [16]. It can cause allergic contact dermatitis and subsequent facial edema, itching, redness and scaling [7,17].…”

Safety Evaluation and Anti-wrinkle Effects of Retinoids on Skin

“…A proliferative responses of LNC in rats (Arts et al , 1996) , mice (Jung et al , 2010) and guinea pigs (Kashima et al , 1996) were assessed by a non-radioisotope method using bromodeoxyuridine (BrdU) . Recently, cytokine measurement (Hariya et al , 1999 ; Pichowski et al , 2000 ; Ulrich et al , 1998) or phenotype analysis using flow cytometry (Gerberick et al , 1999 ; Jung et al , 2010 ; Manetz and Meade, 1999 ; Sikorski et al , 1996) were performed to supplement the cell proliferation data. It is proposed that IMDS, used in this study, is considered to be a good method amending the defects of LLNA.…”

Development and utilization of an ex vivo bromodeoxyuridine local lymph node assay protocol for assessing potential chemical sensitizers

“…In 1999, the Interagency Coordinating Committee for the Validation of Alternative Methods (ICCVAM) recommended the development and utilization of alternative LLNA endpoints (ICCVAM, 1999). One area selected for improvement involves the development of an LLNA protocol that does not require the use of radioisotope (Idehara et al, 2008;Jung et al, 2010;Kojima et al, 2011;Piccotti et al, 2006;Suda et al, 2002;Takeyoshi et al, 2001Takeyoshi et al, , 2003Takeyoshi et al, , 2005Ulker et al, 2013). A promising approach is to replace 3HTdR with its non-radioactive analog, 5-bromo-2′-deoxyuridine (BrdU).…”

“…Like 3HTdR, BrdU is incorporated into the DNA of locally proliferating lymphocytes during the S-phase of the cell cycle and has been used as a substitute for 3HTdR in many cellular proliferation assays (Porstmann et al, 1985). Because BrdU is similar to 3HTdR, assay variations that utilize BrdU incorporation are very similar to the standard LLNA (Jung et al, 2010) and are gaining acceptance by the international community (e.g., ICCVAM, OECD, ECVAM). Compared to those using 3HTdR, BrdU-based detection systems offer many advantages: elimination of radioisotope (and associated use restrictions); elimination of complicated handling and disposal of radioactive animals and contaminated cages/bedding; and facilities to conduct additional evaluation of isolated lymph node cells (e.g., counting, flow cytometry and cytokine production).…”