Responses to the Standard for Exchange of Nonclinical Data (SEND) in non-US countries

The Standard for the Exchange of Nonclinical Data (SEND), adopted by the US FDA, is part of a set of regulations and guidances requiring the submission of standardized electronic study data for nonclinical and clinical data submissions. SEND is the nonclinical implementation of SDTM (Study Data Tabulation Model), the standard electronic format for clinical regulatory submissions to FDA. SEND, SDTM, and the associated Controlled Terminology have been developed by CDISC (Clinical Data Interchange Standards Consortium). In order to successfully implement SEND, interdisciplinary contributions between sponsors and CROs, need a model for task allocation. This is being undertaken by the Pharmaceutical Users Software Exchange (PhUSE). Because SEND is currently the preferred submission format of the US FDA only and will become required by it starting in December 2016, only American academic societies and companies are actively involved. An exception to this is the INHAND initiative, which leads the way in standardizing terminology for toxicological pathology. On the other hand, international globalization of other clinical and nonclinical practices is not feasible because there are substantial differences between the US and non-US countries in CRO involvement in drug development. Thus, non-US countries must consider and develop approaches to SEND that meet their needs. This paper summarizes the activities of the major organizations involved in SEND development and implementation, discusses the effective use of SEND, and details a compliance scheme (research material of the Showa University School of Medicine) illustrating how pharmaceutical companies can complete a large amount of work up to an FDA application with the effective utilization of CROs and solution providers.     

Evaluation of Cold Tolerance in NERICAs Compared with Japanese Standard Rice Varieties at the Reproductive Stage

New Rice for Africa (NERICA) is a general name for interspecific rice varieties derived from a cross between the high‐yielding Asian rice (Oryza sativa L.) between locally adapted African rice (Oryza glaberrima Steud.). Eight NERICAs were evaluated for cold tolerance (CT) at the reproductive stage and compared with their O. sativa parents and three Japanese standard rice varieties over 3 years. Cold tolerance was evaluated based on the filled grain ratio (FGR) after cold water irrigation. The FGR was greatly reduced by cold water irrigation. NERICA 1, 2 and 7 had higher FGR (51.9–57.9 %), while NERICA 6, 15 and 16 had lower FGR (6.2–14.5 %). NERICA 1, 2 and 7 were less affected by cold stress, with a 31 % mean reduction in FGR, while NERICA 6, 15 and 16 were greatly affected, with their FGRs being reduced by more than 80 %. NERICA 3 and 4 were moderately affected by cold stress, with about 45 % reduction rate in FGR. FGR significantly influenced the grain weights of the varieties with strong positive correlations (r = 0.83–0.91; P < 0.001), and thus, similar trends in grain weights were observed. Grain weights were reduced by 61.7–96.4 % under cold stress. NERICA 1, 2 and 7 showed significantly better performance than NERICA 3 and 4, while NERICA 6, 15 and 16 performed poorly under cold water irrigation. The Japanese varieties Koshihikari (very tolerant) and Ozora (moderately tolerant) were more affected by cold water irrigation than NERICA 1, 2 and 7. On the basis of the mean reduction rate (%) in FGR under cold stress, the varieties were classified as follows: NERICA 1, 2 and 7 as tolerant; NERICA 3 and 4 as moderately tolerant; and NERICA 6, 15 and 16 as susceptible to cold stress. However, NERICA 7 grain yields were lower under cold stress due to both greatly reduced number of panicles per plant and number of spikelets per panicle. Therefore, NERICA 1 and 2 are suitable candidates for production in the highland regions of East Africa and should be promoted for production.     

Characterization of genes for novel thaumatin-like proteins in Cryptomeria japonica

Thaumatin-like proteins (TLPs) are induced by a variety of phytopathogens in many plants and several TLPs are allergenic. Previously, we isolated three TLP-encoding cDNAs (Cry j 3.1, Cry j 3.2 and Cry j 3.3) from a cDNA library derived from the pollen of Cryptomeria japonica D. Don. Here, we describe three new TLP cDNAs (Cry j 3.4, Cry j 3.5 and Cry j 3.6). We compared the sequences, the genetic map location and the expression patterns of the Cry j 3 genes. The amino acid sequence predicted from Cry j 3.5 exhibits only limited similarity to those predicted from the other Cry j 3 genes. Linkage analysis showed that the Cry j 3.1 to Cry j 3.4 genes are located in the same linkage group, but Cry j 3.5 is located in a different group. Organ-specificity and induction by stresses and plant hormones differed among the Cry j 3 mRNAs. In pollen grains, the Cry j 3.5 mRNA expression level was higher than that of the other Cry j 3 genes. Exposure to UV-B and salt stress induced expression of Cry j 3.1. The ethylene-releasing compound ethephon strongly induced expression of Cry j 3.4. Salt stress and salicylic acid also induced expression of Cry j 3.4. Abscisic acid weakly induced expression of Cry j 3.5. Arachidonic acid strongly induced expression of Cry j 3.4 and Cry j 3.6, and weakly induced that of Cry j 3.3, whereas expression of Cry j 3.1 and Cry j 3.5 was unaffected. These results suggest that the roles of TLPs and the cascades that regulate their expression differ among the members of the TLP family in C. japonica.     

Estimating the production and mortality of fine roots in a Japanese cedar (Cryptomeria japonica D. Don) plantation using a minirhizotron technique

Fine roots are a key component of forested ecosystems, but available information is still limited. This study examined the production and mortality of fine roots less than 1 mm in diameter in a Japanese cedar (Cryptomeria japonica D. Don) plantation located on the Kanto Plain in central Japan. We used a minirhizotron technique in combination with soil coring, and collected data for 1 year (May 2002–May 2003). Fine root production and mortality were determined from changes in the lengths of individual fine roots on minirhizotron tubes. Both fine root production and mortality rates were greater in the upper soil than in lower soil levels. Both rates were seasonal, with higher values in summer than in winter; this trend was more pronounced in upper soil levels. These results suggest that environmental conditions, such as temperature or soil properties, affect the production and mortality rates of fine roots. Fine root production and mortality occurred simultaneously, and their rates were similar, which may have led to unclear seasonal changes in fine root standing crop estimates. Soil coring indicated that the fine root biomass of this stand was about 120 g m⁻², of which 40% was from Japanese cedar. The estimated rates of dry matter production and mortality of total fine roots, including understory plants, were both approximately 300 g m⁻² year⁻¹.     

Nitrogen-fixing activity in decomposing litter of three tree species at a watershed in eastern Japan

Nitrogen fixation during litter decomposition was studied for 34 months using litterbags containing newly fallen litter of coniferous species Cryptomeria japonica and Pinus densiflora and that of deciduous species Quercus serrata. Litterbags were set in contact with the forest floor in a deciduous broad-leaved forest near the top of a slope and in a C. japonica stand at the middle of the slope at a watershed in eastern Japan. Nitrogen-fixing activity, estimated by acetylene reduction after 16 and 19 months of incubation, was 62.65–3.86 nmoles C₂H₄ h⁻¹ g⁻¹ DW in Cryptomeria litter, but only 1.07–0.09 in Pinus and 0.72–0.04 in Quercus. The rate of N increase in decomposing litter was highest in Cryptomeria. Fungal biomass in decomposing litter, estimated by ergosterol content, increased during the initial 16 months of incubation in Cryptomeria and Quercus, and during the initial 19 months of incubation in Pinus. The litter decomposition rate was highest in Cryptomeria among the three species, due to increased N content and fungal biomass in Cryptomeria litter. Thus, N increase in decomposing Cryptomeria litter affects the subsequent N dynamics and decomposition pattern.     

Internal Distribution of Acid Materials within Snow Crystals

We found that river water is acidified not only in the first stage but also in the later stage of the snowmelt season in Japan, which differs from the so called acid shock occurring in the first stage of the snowmelt in the northern Europe. The acid shocks depend on the regional characteristics of the melt-refreeze processes forming the internal distribution of acid materials within snow crystals under the warm metamorphism. In a warm climatic region like the central Japan, there are possibilities to have the complicated distribution structure of acid materials within granular-snow crystals due to the repeated melt-refreeze processes even in midwinter. Consequently, the pH value of meltwater does not always increase as the snowmelt proceeds. Then, we showed the possibility by using the X-ray computed tomography that the domains with the acid materials exist in the inner parts of snow crystals. So, the acidification of the river water may occur even in the later stage of the snowmelt.     

Induction of apoptotic lesions in liver and lymphoid tissues and modulation of cytokine mRNA expression by acute exposure to deoxynivalenol in piglets

Six 1-month-old piglets were intravenously injected with deoxynivalenol (DON) at the concentration of 1 mg/kg body weight, with three pigs each necropsied at 6 and 24 h post-injection (PI) for investigation of hepatotoxicity and immunotoxicity with special attention to apoptotic changes and cytokine mRNA expression. Histopathological examination of the DON-injected pigs revealed systemic apoptosis of lymphocytes in lymphoid tissues and hepatocytes. Apoptosis of lymphocytes and hepatocytes was confirmed by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and immunohistochemical staining against single-stranded DNA and cleaved caspase-3. The number of TUNEL-positive cells in the thymus and Peyer''s patches of the ileum was increased at 24 h PI compared to 6 h PI, but the peak was at 6 h PI in the liver. The mRNA expression of interleukin (IL)-1β, IL-6, IL-18, and tumor necrosis factor (TNF)-α in the spleen, thymus and mesenteric lymph nodes were determined by semi-quantitative RT-PCR, and elevated expression of IL-1β mRNA at 6 h PI and a decrease of IL-18 mRNA at 24 h PI were observed in the spleen. IL-1β and IL-6 mRNA expressions increased significantly at 6 h PI in the thymus, but TNF-α decreased at 6 h PI in the mesenteric lymph nodes. These results show the apoptosis of hepatocytes suggesting the hepatotoxic potential of DON, in addition to an immunotoxic effect on the modulation of proinflammatory cytokine genes in lymphoid organs with extensive apoptosis of lymphocytes induced by acute exposure to DON in pigs.     

First report of blueberry red ringspot disease caused by <Emphasis Type="Italic">Blueberry</Emphasis><Emphasis Type="Italic">red</Emphasis><Emphasis Type="Italic">ringspot</Emphasis><Emphasis Type="Italic">virus</Emphasis> in Japan

Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush blueberry (Vaccinium corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome. This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan. The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884 to AB469893 for BRRV isolates from Japan.