Abstract A mixture of four squalestatins was separated anil characterized by gradient packed capillary liquid chromatography combined with electrospray collision‐induced dissociation mass spectrometry (ESI/CID/MS). Negative‐ion ESI was found to afford spectra typified by strong pseudo‐molecular ions but providing little structural information. Structural information was obtained using an ion‐focusing RF‐only octapole lens to provide conditions for CID. The new method showed improvements in sensitivity when compared to existing thermospray and dynamic liquid secondary‐ion mass spectrometry methods.

Intercrosses within three I 1 families of red clover ( Trifolium pratense L.) established that two families possessed both of the gametophytic homozygous S‐allele classes (S 1 S 1 and S 2 S 2 ) along with the heterozygous class (S 1 S 2 ). The other family segregated only one homozygous class (S 1 S 1 ) indicating the possibility of differential growth rates of pollen tubes. Reciprocal backcrosses of 15 I 1 progenies to their I 0 clones revealed a deficiency of the sum of the two homozygous classes. The inference is that the deficiency may have resulted from heterozygotic advantage or from zygotic lethality. We conclude that the gametophytic type of S‐allele system is common in red clover, but segregation of S‐alleles is influenced by the interaction of alleles with temperature and possibly other environmental factors.

The rat hepatic stellate cell line PAV-1 was established two decades ago and proposed as a cellular model to study aspects of hepatic retinoic acid metabolism. This cell line exhibits a myofibroblast-like phenotype but also has the ability to store retinyl esters and synthesize retinoic acid from its precursor retinol. Importantly, when cultured with palmitic acid alone or in combination with retinol, the cells switch to a deactivated phenotype in which the proliferation and expression of profibrogenic marker genes are suppressed. Despite these interesting characteristics, the cell line has somehow fallen into oblivion. However, based on the fact that working with in vivo models is becoming increasingly complicated, genetically characterized established cell lines that mimic aspects of hepatic stellate cell biology are of fundamental value for biomedical research. To genetically characterize PAV-1 cells, we performed karyotype analysis using conventional chromosome analysis and multicolor spectral karyotyping (SKY), which allowed us to identify numerical and specific chromosomal alteration in PAV-1 cells. In addition, we used a panel of 31 species-specific allelic variant sites to define a unique short tandem repeat (STR) profile for this cell line and performed bulk mRNA-sequencing, showing that PAV-1 cells express an abundance of genes specific for the proposed myofibroblastic phenotype. Finally, we used Rhodamine-Phalloidin staining and electron microscopy analysis, which showed that PAV-1 cells contain a robust intracellular network of filamentous actin and process typical ultrastructural features of hepatic stellate cells.

The cultivated clovers ( Trifolium spp.) are susceptible to a wide diversity of pests, and potential vulnerability exists in part because of a lack of programs that monitor crop‐pest relationships. The trend in number and diversity of cultivars of the cultivated species is downward, perhaps because of emphasis on other crops and narrowing of gene bases by breeding. However, because the cultivated clovers are mostly cross pollinated, heterozygous, and heterogeneous we do not believe they are particularly vulnerable at present. Germplasm resources are believed to be in a more hazardous position. Although many accessions of the cultivated species have been assembled at USDA Regional Plant Introduction stations and by scientists of other agencies, the accessions have not been examined and cataloged extensively for genetic characters including pest resistance. Only a few germplasm pools are now being maintained in the cultivated species. A complete collection of the wild Trifolium species has not been assembled, and many of those once available have been lost. It is recommended that collection of species and ecotypes of Trifolium be started immediately so that variability in the genus can be preserved and analyzed. Phylogenetic studies, including interspecific hybridization in the genus, should be expanded. Methods of evaluation and use of accessions should be established by a committee of experts for the Trifolium genus so that recommendations for action can be made to a national coordinating committee concerned with germplasm resources.

Synopsis The size of red clover plants which grew under laboratory conditions was not directly correlated with persistence of plants of the same variety in the field. The smallest plants obtained in the controlled environment were of the most persistent variety. The varieties did not differ significantly when the plants were subjected to a frequent‐clipping treatment or prolonged incubation in a dark chamber.

Genetic control of crossing was accomplished by using inbreds with homozygous S‐alleles to produce double‐cross hybrid red clover ( Trifolium pratense L.). The method, checked by the use of the leafmark gene, proved to be effective in controlling crosses for the production of hybrids. Single and double crosses were evaluated for performance. Certain I 1 single and double crosses showed consistently good performance in all characters studied, but most were not superior to the best check variety. Significant correlations were observed between characters evaluated in spaced plantings and broadcast plots, indicating that spaced plantings can be used effectively in the preliminary selection of superior single and double crosses.

Perilipin 5 (PLIN5) is a lipid droplet coat protein that is highly expressed in oxidative tissues such as those of muscles, the heart and the liver. PLIN5 expression is regulated by a family of peroxisome proliferator-activated receptors (PPARs) and modulated by the cellular lipid status. So far, research has focused on the role of PLIN5 in the context of non-alcoholic fatty liver disease (NAFLD) and specifically in lipid droplet formation and lipolysis, where PLIN5 serves as a regulator of lipid metabolism. In addition, there are only limited studies connecting PLIN5 to hepatocellular carcinoma (HCC), where PLIN5 expression is proven to be upregulated in hepatic tissue. Considering that HCC development is highly driven by cytokines present throughout NAFLD development and in the tumor microenvironment, we here explore the possible regulation of PLIN5 by cytokines known to be involved in HCC and NAFLD progression. We demonstrate that PLIN5 expression is strongly induced by interleukin-6 (IL-6) in a dose- and time-dependent manner in Hep3B cells. Moreover, IL-6-dependent PLIN5 upregulation is mediated by the JAK/STAT3 signaling pathway, which can be blocked by transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α). Furthermore, IL-6-mediated PLIN5 upregulation changes when IL-6 trans-signaling is stimulated through the addition of soluble IL-6R. In sum, this study sheds light on lipid-independent regulation of PLIN5 expression in the liver, making PLIN5 a crucial target for NAFLD-induced HCC.

The Trifolium species of eastern North America now occupy a much reduced area in comparison to their earlier range. These species include T. reflexum L. (buffalo clover), T. stoloniferum Muhl. ex Eaton (running buffalo clover), T. carolinianum Michx. (carolina clover), T. virginicum Small ex Small and Vail (Kate's mountain clover), T. bejariense Moric., and a recently discovered species, T. calcarium Collins and Wieboldt. These species all have some potential for use as pasture, and may have desirable qualities for transfer to more widely used forages, such as T. repens L. (white clover). These species were examined to determine morphological and breeding system relationships in comparison with T. repens , and because several are threatened with extinction, to gain information that may optimize management schemes for their long‐term conservation. Based on morphological characteristics, breeding systems, crossing relationships, chromosome numbers, and forage yields, it was concluded that all species are quite distinct and unrelated to T. repens . All the native species had 16 somatic chromosomes. Two annual species, T. carolinianum and T. bejariense , are autogamously self pollinated, as is another annual, T. reflexum , but which may also be cross pollinated to a limited extent in nature. Trifolium stoloniferum set fewer seeds by selfing than by crossing, but self seed set was adequate to maintain the species in nature. Trifolium calcaricum also produced seeds via self‐pollination. One self population of T. stoloniferum produced less biomass than other self and cross populations, suggesting that inbreeding and subsequent loss of vigor may be a possible cause of its near extinction. However, there is insufficient reason to implicate factors other than habitat destruction and competition with other forage species as primary causes of near extinction.