Expression and Functional Relevance of Cannabinoid Receptor 1 in Hodgkin Lymphoma

Background

Cannabinoid receptor 1 (CB₁) is expressed in certain types of malignancies. An analysis of CB₁ expression and function in Hodgkin lymphoma (HL), one of the most frequent lymphomas, was not performed to date.

Design and Methods

We examined the distribution of CB₁ protein in primary cases of HL. Using lymphoma derived cell lines, the role of CB₁ signaling on cell survival was investigated.

Results

A predominant expression of CB₁ was found in Hodgkin-Reed-Sternberg cells in a vast majority of classical HL cases. The HL cell lines L428, L540 and KM-H2 showed strong CB₁-abundance and displayed a dose-dependent decline of viability under CB₁ inhibition with AM251. Further, application of AM251 led to decrease of constitutively active NFκB/p65, a crucial survival factor of HRS-cells, and was followed by elevation of apoptotic markers in HL cells.

Conclusions

The present study identifies CB₁ as a feature of HL, which might serve as a potential selective target in the treatment of Hodgkin lymphoma.     

Geldanamycin-Induced Osteosarcoma Cell Death Is Associated with Hyperacetylation and Loss of Mitochondrial Pool of Heat Shock Protein 60 (Hsp60)

Osteosarcoma is one of the most malignant tumors of childhood and adolescence that is often resistant to standard chemo- and radio-therapy. Geldanamycin and geldanamycin analogs have been recently studied as potential anticancer agents for osteosarcoma treatment. Here, for the first time, we have presented novel anticancer mechanisms of geldanamycin biological activity. Moreover, we demonstrated an association between the effects of geldanamycin on the major heat shock proteins (HSPs) and the overall survival of highly metastatic human osteosarcoma 143B cells. We demonstrated that the treatment of 143B cells with geldanamycin caused a subsequent upregulation of cytoplasmic Hsp90 and Hsp70 whose activity is at least partly responsible for cancer development and drug resistance. On the other hand, geldanamycin induced upregulation of Hsp60 gene expression, and a simultaneous loss of hyperacetylated Hsp60 mitochondrial protein pool resulting in decreased viability and augmented cancer cell death. Hyperacetylation of Hsp60 seems to be associated with anticancer activity of geldanamycin. In light of the fact that mitochondrial dysfunction plays a critical role in the apoptotic signaling pathway, the presented data may support a hypothesis that Hsp60 can be another functional part of mitochondria-related acetylome being a potential target for developing novel anticancer strategies.     

Effects of period RNAi on V-ATPase expression and rhythmic pH changes in the vas deferens of Spodoptera littoralis (Lepidoptera: Noctuidae)

Circadian clocks (oscillators) regulate multiple aspects of insect behaviour and physiology. The circadian system located in the male reproductive tract of Lepidoptera orchestrates rhythmic sperm release from testis and sperm maturation in the upper vas deferens (UVD). Our previous research on the cotton leafworm, Spodoptera littoralis, suggested rhythmic changes in the V-ATPase levels in the UVD epithelium, which correlated with rhythmic pH fluctuations in the UVD lumen. However, it was not known whether UVD cells contain clock mechanism that generates these daily fluctuations. In the current paper, we show circadian rhythm in the expression of clock gene period at the mRNA and protein level in the UVD epithelium. To determine the role of PER in V-ATPase and pH regulation, testes–UVD complexes were treated in vitro with double-stranded fragments of per mRNA (dsRNA). This treatment, which transiently lowered per mRNA and protein in the UVD, altered expression of V-ATPase c subunit. In addition, per RNAi caused a significant delay in the UVD lumen acidification. These data demonstrate that the UVD molecular oscillator involving the period gene plays an essential role in the regulation of rhythmic V-ATPase activity and periodic acidification of the UVD lumen.     

Mammary invasive ductal adenocarcinoma and vulvar invasive keratinizing squamous cell carcinoma in a woman with disseminated discoid lupus erythematosus (DDLE) - a case review

A case of metachronous presentation of two malignancies, invasive mammary ductal adenocarcinoma and later invasive keratinizing squamous cell vulvar carcinoma, complicating a 20 years long course of disseminated discoid lupus erythematosus is described. The authors underline a potential role of long-standing immunotherapy as a factor responsible for inducing secondary neoplasia.     

Successful cryopreservation of Quercus robur plumules

Successful cryopreservation of Q. robur germplasm as plumules (i.e. shoot apical meristems of embryos) is described in this paper. After excision from the recalcitrant seeds and preliminary storage in 0.5 M sucrose solution (18 h), the plumules were subjected to cryoprotection (in 0.75 M sucrose, followed by 1.0 M sucrose and 1.5 M glycerol solutions), and next to desiccation (over silica gel or in nitrogen gas) and cooling (in slush at –210°C or in vials filled with liquid nitrogen, LN, −196°C), and were then cryostored for 24 h. High percentage of survival was obtained after cryostorage (21–67%, depending on pretreatment, assessed in vitro by greening plumules that increased in size). Desiccation of plumules over silica gel resulted in significantly higher survival after cryopreservation (58%) in comparison with desiccation in nitrogen gas (29%), with regrowth (shoots with leaves) 5–18%. The extent of plumule desiccation was comparable in both methods, in which drying of plumules for 20 min decreased the water content to 0.5–0.6 g H₂O g⁻¹ dry weight before LN exposure. The type of LN exposure did not significantly influence plumule survival and regrowth after cryostorage. Plumules isolated from acorns of four provenances survived cryostorage after cryoprotection followed by desiccation over silica gel and direct cooling in vials with LN (survival 51–76%, regrowth 8–20%). Normal plants developed from the recovered shoots after rooting. The presented protocol for Q. robur plumule cryopreservation may offer a potential approach for establishing germplasm conservation in gene banks for Quercus species.     

Induction of aromatic ring: cleavage dioxygenases in <Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis> strain KB2 in cometabolic systems

Stenotrophomonas maltophilia KB2 is known to produce different enzymes of dioxygenase family. The aim of our studies was to determine activity of these enzymes after induction by benzoic acids in cometabolic systems with nitrophenols. We have shown that under cometabolic conditions KB2 strain degraded 0.25–0.4 mM of nitrophenols after 14 days of incubation. Simultaneously degradation of 3 mM of growth substrate during 1–3 days was observed depending on substrate as well as cometabolite used. From cometabolic systems with nitrophenols as cometabolites and 3,4-dihydroxybenzoate as a growth substrate, dioxygenases with the highest activity of protocatechuate 3,4-dioxygenase were isolated. Activity of catechol 1,2- dioxygenase and protocatechuate 4,5-dioxygenase was not observed. Catechol 2,3-dioxygenase was active only in cultures with 4-nitrophenol. Ability of KB2 strain to induce and synthesize various dioxygenases depending on substrate present in medium makes this strain useful in bioremediation of sites contaminated with different aromatic compounds.     

Comparative analysis of hydrogen-producing bacterial biofilms and granular sludge formed in continuous cultures of fermentative bacteria

A system for biohydrogen production was developed based on long-term continuous cultures grown on sugar beet molasses in packed bed reactors. In two separate cultures, consortia of fermentative bacteria developed as biofilms on granitic stones. In one of the cultures, a granular sludge was also formed. Metagenomic analysis of the microbial communities by 454-pyrosequencing of amplified 16S rDNA fragments revealed that the overall biodiversity of the hydrogen-producing cultures was quite small. The stone biofilm from the culture without granular sludge was dominated by Clostridiaceae and heterolactic fermentation bacteria, mainly Leuconostocaeae. Representatives of the Leuconostocaeae and Enterobacteriaceae were dominant in both the granules and the stone biofilm formed in the granular sludge culture. The culture containing granular sludge produced hydrogen significantly more effectively than that containing only the stone biofilm: 5.43 vs. 2.8 mol H₂/mol sucrose from molasses, respectively. The speculations that lactic acid bacteria may favor hydrogen production are discussed.     

Frequency of detection of Ureaplasma urealyticum and Mycoplasma hominis in cervical canal and the Douglas pouch of infertile and fertile women

The group of organisms commonly referred to as genital mycoplasmas comprise species most often found in genitourinary tract of sexually active adults as common commensal inhabitants, or pathogens which can possibly cause many different pathologies like: non-gonococcal urethritis, bacterial vaginosis, cervicitis, endometritis or pelvic inflammatory disease. The problem of their morbidity and the possible influence they have on human fertility is still not clear. The aim of this study was to find out whether two investigated species- Ureaplasma urealyticum and Mycoplasma hominis can be detect more often in a group of infertile women. 74 women participated in the study and were assigned to one of 2 groups of patients: infertile women and fertile women without any sign of genital tract infection. Swabs from the cervical canal of the uterus and the fluid from the Douglas pouch were taken during the gynecological examination and laparoscopic procedure. Two diagnostic methods were used: biochemical method- commercial diagnostic kit- Mycoplasma IST 2 and PCR method. The results showed that Ureaplasma urealyticum and Mycoplasma hominis were detected among both fertile and infertile women with nearly the same frequency, much more often in cervical canal than in the Douglas pouch. Ureaplasma urealyticum was more common pathogen than Mycoplasma hominis in both groups and locations. The achieved results point out that the role of genital mycoplasmas in human infertility is still unclear and require further investigations.     

Liming to remediate Ni contaminated soils with diverse properties and a wide range of Ni concentration

Historic emissions from a Ni refinery at Port Colborne, Ontario, caused Ni contamination of regional soils and raised concerns about potential Ni phytotoxicity. Previous tests revealed that if these soils were made alkaline and fertilized with Mn and other common nutrients as needed to maintain fertility of such alkaline soils, full remediation (prevention of Ni phytotoxicity) would be obtained. This experiment was conducted to test this method of remediation on diverse soils from Port Colborne, and to evaluate chemical extraction tests which would be predictive of plant uptake and potential for Ni phytotoxicity in Ni-contaminated soils. Ten soils with varied levels of Ni contamination and varied soil properties were amended with limestone or nitric acid to raise or lower pH so that a wide pH range could be examined for the soils. For lower Ni organic and mineral soils near the Ontario remediation limit (200 mg/kg), neither crop suffered Ni phytotoxicity at any pH tested. Only when more highly contaminated soils were strongly acidic did Ni phytotoxicity occur. Phytotoxic soils were fully remediated by making soils alkaline even for these Ni-sensitive crop species. Only the most contaminated organic soil remained slightly toxic – but this soil was remarkably contaminated (over 1.1% of Ni). The Sr nitrate extraction method was much more effective in predicting plant Ni concentrations than the DTPA method. This test provides an inexpensive soil extraction result highly predictive of potential for Ni phytotoxicity across soils.