Haemochromatosis gene (HFE) mutations in viral-associated neoplasia: Linkage to cervical cancer

The present study examines the frequency of the two main HFE mutations (C282Y and H63D) in a randomly selected population of 346 individuals including 201 DNA samples from women with cervical neoplasia (including high-grade squamous intraepithelial lesions and invasive squamous cell carcinoma) and a control population of 146 women from the same geographical area. We found a significantly lower risk of development of cervical neoplasia in H63D carriers (OR = 0.56; 95% CI 0.35-0.92; p = 0.01). Multivariate logistic regression analysis confirms this observation (OR = 0.55; 95% CI 0.35-0.88, p = 0.01). Regarding the C282Y mutation no association was found (OR = 1.32; 95% CI 0.53-3.33; p = 0.52). In addition, a significant difference between H63D carrier and non-carrier women on the time-to-onset of cervical lesions was observed (log-rank test: p = 0.0012). These results indicate that HFE could be considered a candidate modifier gene of viral-related neoplasia such as cervical carcinoma possibly by a dual role on iron metabolism and immunological system.     

Contribution to the occurrence and former distribution of Tephroseris palustris (Compositae) in the Central Europe

Tephroseris palustris (syn. Senecio palustris) is a circumboreal species with large distribution range. The European part of the recent distribution area extends southwards to central France, Germany, Poland, and Ukraine, while in Great Britain, Czech Republic, Hungary, Slovakia, and Romania, T. palustris has been treated as extinct species. The southern boundary of its distribution in Poland does not reach the Carpathian territory. Herbarium specimens, formerly collected in Czech Republic, were found, however, all Czech localities are extinct. No herbarium specimens confirming the old literature data from Slovakia, Hungary, and Romania have been found. Some herbarium specimens coming from this area, and declared as T. palustris (S. palustris), in fact, refer to Senecio paludosus L. Contrary to previous nomenclature review (Jeffrey & Chen 1984), the name Tephroseris palustris (L.) Rchb. seems to be correct (Reichenbach Fl. Saxon.: 146, 1842).     

Ependyma as a Possible Morphological Basis of Ischemic Preconditioning Tolerance in Rat Spinal Cord Ischemia Model: Nestin and Fluoro-Jade B Observations

1. To test our hypothesis that a transient nonlethal ischemic insult benefits the lumbosacral spinal cord ischemic injury, nestin, the marker of proliferating cells, and Fluoro-Jade B, the marker of degenerating cells, were used in rats. Morphological outcome was evaluated after 12-min ischemia versus 12-min ischemia preconditioned by 3-min ischemic period and 30-min recirculation (IPC), in each group followed by 2, 3, and 4 days of posttreatment survival. 2. Twelve-minute ischemia, inducing nestin-positivity in ependyma and reactive astrocytes at the L(1-3) spinal cord segments, shows this region as the viable region of spinal cord in all postischemic survival periods. On the other hand, abundance of Fluoro-Jade B-positive cells, distributed throughout the dorsal horn and intermediate zone of L4-S2 segments, points out the most injured spinal cord region by ischemia. 3. After the same ischemic insult in IPC rats only a few nestin-positive ependymal cell and reactive astrocytes appeared beside the nestin-positive vessels in the lower lumbar and sacral spinal cord segments of all survival periods. The appearance of nestin-positive cells in the spinal cord segments, which "should have been affected" by ischemia indicates protection of this region by the IPC treatment. 4. The number and density evaluation of Fluoro-Jade B fluorescent cells of L4-S2 segments after ischemia and IPC confirmed that degenerating cells were significantly reduced in the IPC rats in all survival periods. 5. Our results showing the immunohistochemical response of epemdyma, committed to the presence of viable tissue, indicate that the ependymal cells may contribute to the ischemic resistance in the IPC rats.     

Genetic structure and phylogeography of a temperate-boreal herb, Cardamine scutata (Brassicaceae), in northeast Asia inferred from AFLPs and cpDNA haplotypes

? Premise of the study: Studies on genetic structure of plant populations help us understand the history of local flora and vegetation. In this study, we focus on the temperate-boreal herb Cardamine scutata from northeast Asia, an area with scarce phylogeographic studies. We explore patterns of genetic variation within this species, with an aim to infer its (post-) glacial history with reference to colonization routes and migrations via land bridges. ? Methods: We analyzed 46 populations sampled in Japan, Kamchatka, and Korea using AFLP and cpDNA sequence data. ? Key results: Two intraspecific genetic groups were resolved, distributed in the northeastern and southwestern part of the study area, most likely reflecting lineages isolated from each other during (at least) the last glaciation. A zone of secondary contacts was found in central/northern Honshu, and a few cases of long-distance dispersal were observed. We detected efficient gene flow across the marine straits, supporting the role of land bridges created by sea level decline during the last glacial period. The cpDNA data indicated extensive recent expansion and diversification within both lineages. We inferred recent colonization of Kamchatka from Hokkaido, associated with genetic impoverishment. ? Conclusions: The pattern of north-south genetic differentiation found in C. scutata is rather common among several other plant species studied in Japan, despite their distinct biological features. We assume that different processes and factors may have brought about this similarity. Overall, this study contributes to better understanding of the biogeography of northeast Asia.     

Chinese hamster ovary (CHO) host cell engineering to increase sialylation of recombinant therapeutic proteins by modulating sialyltransferase expression

N‐Glycans of human proteins possess both α2,6‐ and α2,3‐linked terminal sialic acid (SA). Recombinant glycoproteins produced in Chinese hamster overy (CHO) only have α2,3‐linkage due to the absence of α2,6‐sialyltransferase (St6gal1) expression. The Chinese hamster ST6GAL1 was successfully overexpressed using a plasmid expression vector in three recombinant immunoglobulin G (IgG)‐producing CHO cell lines. The stably transfected cell lines were enriched for ST6GAL1 overexpression using FITC‐Sambucus nigra (SNA) lectin that preferentially binds α2,6‐linked SA. The presence of α2,6‐linked SA was confirmed using a novel LTQ Linear Ion Trap Mass Spectrometry (LTQ MS) method including MSn fragmentation in the enriched ST6GAL1 Clone 27. Furthermore, the total SA (mol/mol) in IgG produced by the enriched ST6GAL1 Clone 27 increased by 2‐fold compared to the control. For host cell engineering, the CHOZN^® GS host cell line was transfected and enriched for ST6GAL1 overexpression. Single‐cell clones were derived from the enriched population and selected based on FITC‐SNA staining and St6gal1 expression. Two clones (“ST6GAL1 OE Clone 31 and 32”) were confirmed for the presence of α2,6‐linked SA in total host cell protein extracts. ST6GAL1 OE Clone 32 was subsequently used to express SAFC human IgG1. The recombinant IgG expressed in this host cell line was confirmed to have α2,6‐linked SA and increased total SA content. In conclusion, overexpression of St6gal1 is sufficient to produce recombinant proteins with increased sialylation and more human‐like glycoprofiles without combinatorial engineering of other sialylation pathway genes. This work represents our ongoing effort of glycoengineering in CHO host cell lines for the development of “bio‐better” protein therapeutics and cell culture vaccine production. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:334–346, 2015