A new continuous cell line from larval ovaries of silkworm, <Emphasis Type="Italic">Bombyx mori</Emphasis>

A new continuous cell line from ovarian tissue of commercial variety “Kolar Gold” of silkworm, Bombyx mori, was established and designated as DZNU-Bm-12. The tissue was grown in MGM-448 insect cell culture medium supplemented with 10% fetal bovine serum (FBS) and 3% heat-inactivated B. mori hemolymph at 25 ± 1°C. The migration of partially attached small round refractive cells from the fragments of ovarioles began from the beginning of explantation. The cells multiplied partially attached in the primary culture initially, and some of them become freely suspended after 20 passages. The cells were adapted to MGM-448 and TNM-FH media each with 10% FBS and the population doubling time of cell line was about 36 and 24 hr, respectively. The chromosome number was near diploid at initial passages and slightly increased at 176th passage, but a few tetraploids and hexaploids were also observed. DNA profiles using simple sequence repeat loci established the differences between DZNU-Bm-12 and DZNU-Bm-1 and most widely used Bm-5 and BmN cell lines. The cell line was found susceptible to B. mori nucleopolyhedrovirus (BmNPV) with 85–90% of the cells harboring BmNPV and having an average of 3–17 OBs/infected cell. We suggest the usefulness of this cell line in BmNPV-based baculoviral expression system and also for studying in vitro virus replication.     

Modulation of intestinal urea cycle by dietary spermine in suckling rat

Argininosuccinate synthetase, an ubiquitous enzyme in mammals, catalyses the formation of argininosuccinate, the precursor of arginine. Arginine is recognised as an essential amino acid in foetuses and neonates, but also as a conditionally essential amino acid in adults. Argininosuccinate synthetase is initially expressed in enterocytes during the developmental period, it disappeared from this organ then appeared in the kidneys. Although the importance of both intestinal and renal argininosuccinate synthetases has been recognised for a long time, nutrients have not yet been identified as inducers of the gene expression. In the context of a proteomic screening of intestinal modifications induced by dietary spermine in suckling rats, we showed that argininosuccinate synthetase and carbamoyl phosphate synthase disappeared from enterocytes after this treatment. The disappearance of argininosuccinate synthetase in small intestine was confirmed by immunodetection. Expression of carbamoyl phosphate synthase and argininosuccinate synthetase coding genes decreased also after spermine administration. Expression of other urea cycle enzyme coding genes was modulated by spermine administration: argininosuccinate lyase decreased and arginase increased. Our results fit with the developmental variation of argininosuccinate synthetase and carbamoyl phosphate synthase. Modulation of the gene expression for several urea cycle enzymes suggests a coordination between all the pathway steps and switch toward polyamine (or proline and glutamate) biosynthesis from ornithine.     

A preliminary study on ultra high frequency electromagnetic fields effect on black locust chlorophylls

Chlorophylls were quantitatively studied in the leaves of black locust (Robinia pseudoacacia L.) seedlings exposed to electromagnetic fields of high frequency. Exposure system was designed and built up to make possible simultaneous exposure of seedling lots (3 months old) to low power density electromagnetic fields corresponding to a frequency of 400 MHz. After three weeks of daily exposures (1, 2, 3 and 8 hours), chlorophyll levels were measured using adequate spectral device. Statistical analysis of experimental results was performed by means of t-test to identify significant modifications induced by electromagnetic treatment in exposed samples in comparison to the control. Chlorophyll-a as well as chlorophyll-b level was found to decrease except the exposure time of two hours, where a considerable enhancement was noticed. It was revealed that the ratio of the two main types of chlorophyll was decreasing logarithmically to the increase of daily exposure time.     

Determination and validation of a simple high-performance liquid chromatographic method for simultaneous assay of iprodione and vinclozolin in human urine

A method based on solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) was developed for the simultaneous determination of 3-(3,5-diclorophenyl)-5-ethenyl-5-methyl-2,4-oxazolidinedione (vinclozolin) and 3-(3,5-diclorophenyl)-N-(1-methylethyl)-2,4-dioxo-1-imidazolidinecarboxamide (iprodione) in human urine. Urine samples containing vinclozolin and iprodione were collected by solid phase extraction using C(18) cartridges. The chromatographic separation was achieved on a Spherisorb ODS2 (250 mm x 4.6 mm, 5 microm) column with an isocratic mobile phase of acetonitrile-water (60:40, v/v). Detection was UV absorbance at 220 nm. The calibration graphs were linear from 30 to 1000 ng/mL for the two fungicides. Intra- and inter-day R.S.D. did not exceed 2.9%. The quantitation limit was 50 ng/mL for vinclozolin and 30 ng/mL for iprodione, respectively.     

HLA-B57 is significantly associated with psoriasis in Northeast Romania [corrected

The aim of the study was to identify HLA class I types associated with susceptibility to psoriasis among population of Northeast region of Romania. PATIENTS AND METHODS: 27 psoriatic patients and 89 controls were typed serologically for HLA-A and HLA-B lymphocyte expression using CDC-NIH (complement dependent-cytotoxicity--National Institute of Health) method. The Terasaki plates used for HLA class I typing were prepared in our laboratory using antisera of known specificities. RESULTS AND CONCLUSIONS: psoriatic patients in the group of study frequently express HLA-B57 phenotype. The relative risk induced by this phenotype was highly statistically significant (p = 0.0016) while HLA-B13 was not associated with a significant risk for developing psoriasis in patients under study compared with controls (p = 0.48). Also, HLA-B27 (p = 0.96) and HLA-B44 (p = 0.99), reported by others to be associated with late psoriasis, do not meet (a particular) disease susceptibility between psoriatic patients under investigation.     

The Behavioral Response of Larval Amphibians (Ranidae) to Threats from Predators and Parasites

Organisms are exposed to strong selective pressures from several sources, including predators and pathogens. Response to such interacting selective pressures may vary among species that differ in life history and ecology in predictable ways. We consider the impact of multiple enemies (fish predators and trematode parasites) on the behavior of larvae of three anuran species (Lithobates ( = Rana) sylvaticus, L. clamitans and L. catesbeianus). We show that the three ranid species differ in response to the trade-off imposed by the simultaneous presence of fish predators and trematode parasites in the environment. Two more permanent pond breeders (L. clamitans and L. catesbeianus), which commonly encounter parasites and fish, increased activity when in the combined presence of parasites and a fish predator, resulting in a relatively lower parasite encystment rate. In contrast, the temporary pond breeder (L. sylvaticus), which does not commonly encounter fish in the wild, decreased activity in the combined presence of a fish predator and parasites similar to when only the predator was present. For L. sylvaticus, this suggests that the presence of an unknown predator poses a greater threat than parasites. Further, the presence of fish along with parasites increased the susceptibility of both L. sylvaticus and L. clamitans to trematode infection, whereas parasite infection in L. catesbeianus was unaffected by the presence of fish. Unpalatability to fish may allow some species to respond more freely to attacking parasites in the presence of fish. The results from this study highlight the importance of considering multiple selective pressures faced by organisms and how this shapes their behavior.     

The discovery of potent antagonists of NPBWR1 (GPR7)

The synthesis and evaluation of small molecule antagonists of the G protein-coupled receptor NPBWR1 (GPR7) are reported for the first time. [4-(5-Chloropyridin-2-yl)piperazin-1-yl][(1S,2S,4R)-4-{[(1R)-1-(4-methoxyphenyl)ethyl]amino}-2-(thiophen-3-yl)cyclohexyl]methanone (1) emerged as a hit from a high-throughput screen. Examination of substituents that focused on replacing the 5-chloropyridine and 4-methoxybenzylamino groups of 1 led to the identification of compounds that exhibited subnanomolar potencies as low as 660pM (9k) in the functional assay and 200pM in the binding assay (9i).     

Combination of tyrosine kinase inhibitors and the MCL1 inhibitor S63845 exerts synergistic antitumorigenic effects on CML cells

Tyrosine kinase inhibitor (TKI) treatment has dramatically improved the survival of chronic myeloid leukemia (CML) patients, but measurable residual disease typically persists. To more effectively eradicate leukemia cells, simultaneous targeting of BCR-ABL1 and additional CML-related survival proteins has been proposed. Notably, several highly specific myeloid cell leukemia 1 (MCL1) inhibitors have recently entered clinical trials for various hematologic malignancies, although not for CML, reflecting the insensitivity of CML cell lines to single MCL1 inhibition. Here, we show that combining TKI (imatinib, nilotinib, dasatinib, or asciminib) treatment with the small-molecule MCL1 inhibitor {"type":"entrez-nucleotide","attrs":{"text":"S63845","term_id":"400540","term_text":"S63845"}}S63845 exerted strong synergistic antiviability and proapoptotic effects on CML lines and CD34+ stem/progenitor cells isolated from untreated CML patients in chronic phase. Using wild-type BCR-ABL1-harboring CML lines and their T315I-mutated sublines (generated by CRISPR/Cas9-mediated homologous recombination), we prove that the synergistic proapoptotic effect of the drug combination depended on TKI-mediated BCR-ABL1 inhibition, but not on TKI-related off-target mechanisms. Moreover, we demonstrate that colony formation of CML but not normal hematopoietic stem/progenitor cells became markedly reduced upon combination treatment compared to imatinib monotherapy. Our results suggest that dual targeting of MCL1 and BCR-ABL1 activity may efficiently eradicate residual CML cells without affecting normal hematopoietic stem/progenitors.Subject terms: Cancer stem cells, Targeted therapies, Preclinical research