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Rice micronutrients and toxic metal levels are important quality issues impacting “hidden hunger” and food safety. The aim of this study was to evaluate the effects of sowing date (SD) on element accumulations in rice grains. We conducted a rice SD experiment every 7 days from May 10 to June 21 in 2017 and 2018 using 2 japonica (Oryza sativa L.) cultivars. The rice yields were determined, and 19 element contents in the grains were measured via inductively coupled plasma mass spectrometry. Analysis of variance indicated that early SD significantly promoted the rice yields and the micronutrient (i.e., Cu, Fe, Mn, Ni, and Zn) levels in grains. However, early SD also significantly increased Cr and Pb contents but significantly limited Cd accumulations in rice grains. Anyway, the element contents of all determined samples satisfied the requirements of maximum permissible concentrations (GB2762–2012) in this uncontaminated area. Redundancy analysis indicated that temperature and precipitation were the main meteorological drivers of element accumulations. Meanwhile, Cd accumulation was simultaneously affected by the antagonism of Fe/Zn. Thus, we concluded that SD management could be a feasible strategy for promoting micronutrient quality of rice grains and further alleviating Cd accumulations when aiming at high yields.

Velarifictorus ornatus (Shiraki) (Orthoptera: Gryllidae) display distinct wing variation, and a physiological trade-off between reproduction and flight muscles has been observed in long-winged (LW) and short-winged (SW) females. To understand the physiological basis of this trade-off, we investigated the difference in the food digestion capability and digestive enzyme activity between LW and SW females. We found that the efficiency of conversion of digested food into body matter of SW females was significantly higher than that of the LW females, although there was no difference in the approximate digestibility between LW and SW females during the first 12 days after the adult emergence. Similarly, growth and relative growth were significantly higher in SW females. The food consumption of SW females was significantly higher than that of the LW females only in the first 6 days after the adult emergence, suggesting enhanced ovary development in SW females is probably because of the elevated efficiency of conversion of digested food into body matter coupled with greater consumption in the early stage after the adult emergence. Trypsin-like activity was higher in SW females than in LW females, whereas no differences in the fresh weight of the midguts (including content) were observed between LW and SW females at 1, 3, or 5 h after refeeding after starvation for 18 h. In contrast, amylase activity varied significantly depending on time lapse after refeeding, being significantly higher in LW females than in SW females at 1 h after refeeding, but significantly lower in LW females than in SW females at 3 and 5 h after refeeding. The lipase activity displayed a similar trend in both LW and SW females after refeeding, with an initial decline followed by a slight increase; and no difference was observed between LW and SW females at any stage after refeeding.

In wing dimorphic cricket Velarifictorus micado Saussure, group rearing could induce macropters, but rearing density had a nonlinear effect on the incidence of macroptery. Frequency of macropters increased along with rearing density under low-density conditions, but it decreased gradually under high densities, especially for the females. This result indicates that flight ability is required to escape from crowded conditions, but there is a factor that would suppress wing development under high-population density in V. micado. Investigation of body injury showed that incidence of body injury increased along with rearing density, especially for the antenna. Removal of antennas or hindlegs in the penultimate nymphal stage exerted a strong micropterizing effect. These results suggest that brachypterizing effect of high density may be related with body injury. Removed antennas at different instars, brachypterizing effect was only significant in sixth instar (penultimate instar) for males, and sixth and seventh instar (last instar) for females, which suggests that penultimate and last instars may be sensitive stages in winglength development to the effect of body injury in V. micado

Arachnomelia syndrome (AS) is a recessive inherited disease in cattle. Although the arachnomelia phenotypes are virtually identical in Brown Swiss and Simmental cattle, the causative mutation are different, which are a 1 bp insertion c.363-364insG in the sulfite oxidase (SUOX) gene and a 2 bp deletion c.1224_1225delCA in the molybdenum cofactor syn-thesis step 1 (MOCS1) gene, respectively. In the current study, combining fluorescence PCR with capillary electrophoresis technology, an automatic fluorescence method was established, which could detect the two causative loci rapidly and cor-rectly with a single reaction. Samples from 51 Simmental bulls, 80 cows mated artificially using semen of Simmental bulls and their resulted 106 progeny, together with 55 Xinjiang Brown were collected and used for validation of the newly de-signed methods. Our results have laid a foundation for screening AS disease causing mutations in Chinese cattle.

Summary In this study, we assessed the effect of seasonal variation on the semen quality in Chinese by using the average highest temperature (AHT) of the ejaculation day for partitioning season periods. A total of 13 635 semen samples were collected and analysed according to the AHT of the ejaculation day. Semen volume, sperm concentration and the percentage of spermatozoa with normal morphology in midsummer (AHT > 30 °C) were significantly lower than those in other periods of the year. Sperm motility with AHT 10–20 °C in winter and spring was significantly higher than that in other seasons. The percentage of head defects spermatozoa with AHT

We examined the subcellular localization of BRCA1 proteins using immunohistochemical staining with C‐terminal (GLK‐2 antibody) and N‐terminal (Ab‐2 antibody) monoclonal antibodies in 44 familial ovarian cancers. Among these, 24 cases were associated with 13 independent germ‐line mutations of BRCA1, and loss of heterozygosity (LOH) at one or more BRCA1 microsatellite markers was found in all 21 informative tumors tested. With GLK‐2 antibody, cytoplasmic staining was observed in 15 of 16 tumors (93.8%) with mutation in exon 11, and BRCA1 staining was absent in 8 of 8 tumors (100%) with mutation in exons other than exon 11. When immunohistochemical staining was performed with Ab‐2 antibody, both nuclear and cytoplasmic staining were observed in 14 of 16 tumors (87.5%) with mutation in exon 11. Interestingly, nuclear staining was observed in 3 of 3 tumors (100%) with mutation downstream of exon 11, even though no staining was detected in 5 of 5 tumors (100%) with mutation upstream of exon 11. On the other hand, in familial ovarian cancers without BRCA1 mutations, nuclear staining or both nuclear and cytoplasmic staining was observed in 18 of 20 specimens (90%) and 20 of 20 specimens (100%) with GLK‐2 antibody and with Ab‐2 antibody, respectively. These results suggest that an immunohistochemical assay in combination with employing the C‐terminal and the N‐terminal antibodies appears to have potential as a reliable and useful technique for the screening of BRCA1 mutations, at least to predict the status of mutation, upstream or downstream of exon 11.

AIM The genetic region of 6q25 containing the estrogen receptor-alpha (ER-alpha) gene is lost in a significant number of ovarian tumors. The aim of this study was to identify how inherited variation in the ER-alpha gene contributes to susceptibility to familial ovarian cancer. METHODS DNA obtained from 18 cases of familial ovarian cancer without mutation of the BRCA1 and BRCA2 genes, 20 cases with BRCA1 mutation, 20 cases of sporadic ovarian cancer, and 19 controls were screened for mutations in the coding region of the ER-alpha gene using direct sequencing. RESULTS Two germline missense variants at codons 307 [GCC(Ala)-->TCC(Ser)] and 347 [ACC(Thr)-->TCC(Ser)] were detected in two unrelated cases with BRCA1 mutation, but not in all other cases tested. Three polymorphisms in codon 10 [TCT-->TCC(Ser)], codon 325 [CCC-->CCG(Pro)], and codon 594 [ACA-->ACG(Thr)] were identified in this series, and a higher frequency of the allele TCC at codon 10 and a lower frequency of the allele CCG at codon 325 were observed in familial cases without BRCA1 mutation, compared with those in familial cases with BRCA1 mutation, in both the sporadic cases and in the controls. CONCLUSIONS We could not detect pathogenic mutations of the ER-alpha gene in ovarian cancer cases without BRCA1 mutation. However, association analyses of two polymorphisms suggest that the ER-alpha gene or a gene located close to the ER-alpha locus might be related to susceptibility of familial ovarian cancer without BRCA1 mutation.

Human retinal pigment epithelial (RPE) cell cultures are usually obtained from donor eyes; isolation and culture of RPE cells obtained by evisceration has not been reported previously. The present study attempted to isolate and cultivate RPE cells from evisceration specimens obtained from two cases with severe ocular trauma. Two different isolation methods, explantation and enzymatic dissociation, were used. In Case 1, RPE cells grew from the explants, but were contaminated with other cells such as fibroblasts and melanocytes, and no pure RPE cultures were obtained by explantation. In Case 2, RPE cells were separated from choroids using 0.25% trypsin before plating for culture, which effectively eliminated contaminating cells. A pure RPE culture was obtained and cultured with F12 medium supplemented with 30% fetal bovine serum. With this enzymatic dissociation method, cultured RPE cells grew to confluence in primary culture and could be maintained in culture for five passages. Cultured RPE cells were identified by the presence of cytokeratin expression, as shown by immunocytochemical staining. These isolation and culture methods provide alternative sources for human RPE cells and could be useful in studies of the cell biology and pathophysiology of human RPE cells.