Novel human prostate cell lines derived from the transition and peripheral zones of the prostate for carcinogenesis studies

Epithelial cell lines were established from the transition and peripheral zones of human prostate by transduction with cdk4 and hTERT. The properties of these lines were investigated using immunocytochemical markers, ability to generate anchorage-independent colonies and by spectral karyotyping (SKY). Cells were exposed to fractionated doses of gamma irradiation to investigate their ability to transform. Cell lines were established from the transition and peripheral zones of human prostate. The expression of CD133, CK5, CK14, CK18, p16, PSCA, p63 and c-myc varied between the lines from the two regions. The line derived from the peripheral zone exhibited properties of a tumour line. A similar pattern was observed in two separate transductions. It was thus unlikely to be an in vitro transformation event, which is very rarely observed with human cells in vitro, and thus more likely to be derived from the immortalisation of a quiescent tumour clone. Fractionated irradiation of the transition zone cell line resulted in forming of transformed colonies. The transformed and tumour line had marked chromosomal rearrangements as demonstrated by SKY analysis. Cell lines have been derived from different zones of human prostate for studies on radiation carcinogenesis. The unirradiated cell line derived from the peripheral zone exhibited chromosomal rearrangements similar to those observed in prostate carcinoma. The cell line derived from the transitional zone exhibited a near diploid karyotype and could be transformed following exposure to fractionated doses of gamma irradiation.     

Necrosis of the galea as rare complication of infection of a traumatic temporal hematoma

Authors report a case of infective traumatic temporal haematoma causing septic focus. The suppurative inflammation developed two weeks after the trauma causing necrosis of an extensive part of the galea on a big area of the crown of head. A septic process increased producing necrotic alteration of the affected periosteum of the cranial bone and plegmon in the tissue of the neck. Authors report the surgical plastic procedures that were used to establish a granulating layer and to cover the surface in the area of the removed necrotic part of the galea. Authors emphasize the significance of the danger of the inflammatory infiltration and report the effect of the Curiosin solution on the wound-healing.     

Investigation of solubility properties of nifluminic acid containing cyclodextrins and polyvidone

One of the most important task of the pharmaceutical technology is to reach a possible high gastrointestinal absorbtion of the active ingredient. Therefore, it is necessary to have a good solubility in the gastric/intestinal medium. In this way the applied drug quantity and unwanted side effects can be reduced with solubility increasing. The water-insoluble drug's solubility and bioavailability can be increased by the alteration of their physicochemical properties. Nifluminic acid is a frequently used anti-inflammatory drug with low aqueous solubility. Inclusion complexation with cyclodextrins is an efficacious method to improve the solubility, stability and bioavailability. Ternary systems were prepared and investigated to use nifluminic acid, hydoxypropyl-beta-cyclodextrin and polyvidone. The aims of the study were to investigate the solubility rate, the in vitro diffusion ability, to determine the n-octanol/water partition coefficient, and to study the thermoanalitycal properties of products. The results suggested that complexation with cyclodextrin and the use of polyvidone is better method than to prepare binary systems. Inclusion complexation was presumed in the event of the ternary composition to improve the bioavailability of nifluminic acid.     

Neutrophil elastase induces inflammation and pain in mouse knee joints via activation of proteinase‐activated receptor‐2

Background and Purpose

Neutrophil elastase plays a crucial role in arthritis. Here, its potential in triggering joint inflammation and pain was assessed, and whether these effects were mediated by proteinase‐activated receptor‐2 (PAR2).

Experimental Approach

Neutrophil elastase (5 μg) was injected into the knee joints of mice and changes in blood perfusion, leukocyte kinetics and paw withdrawal threshold were assessed. Similar experiments were performed in animals pretreated with the neutrophil elastase inhibitor sivelestat, the PAR2 antagonist GB83, the p44/42 MAPK inhibitor U0126 and in PAR2 receptor knockout (KO) mice. Neutrophil elastase activity was also evaluated in arthritic joints by fluorescent imaging and sivelestat was assessed for anti‐inflammatory and analgesic properties.

Key Results

Intra‐articular injection of neutrophil elastase caused an increase in blood perfusion, leukocyte kinetics and a decrease in paw withdrawal threshold. Sivelestat treatment suppressed this effect. The PAR2 antagonist GB83 reversed neutrophil elastase‐induced synovitis and pain and these responses were also attenuated in PAR2 KO mice. The MAPK inhibitor U0126 also blocked neutrophil elastase‐induced inflammation and pain. Active neutrophil elastase was increased in acutely inflamed knees as shown by an activatable fluorescent probe. Sivelestat appeared to reduce neutrophil elastase activity, but had only a moderate anti‐inflammatory effect in this model.

Conclusions and Implications

Neutrophil elastase induced acute inflammation and pain in knee joints of mice. These changes are PAR2‐dependent and appear to involve activation of a p44/42 MAPK pathway. Blocking neutrophil elastase, PAR2 and p44/42 MAPK activity can reduce inflammation and pain, suggesting their utility as therapeutic targets.

Linked Articles

This article is part of a themed section on Inflammation: maladies, models, mechanisms and molecules. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2016.173.issue-4

Abbreviations

IVM

intravital microscopy

LASCA

laser speckle contrast analysis

PAR

proteinase‐activated receptor

TRPV

transient receptor potential vanilloid

VCAM

vascular cell adhesion molecule

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Changes in surgical therapy of pulmonary hydatid cysts

The lung is the second most common site of hydatid cysts after the liver. The authors analyse retrospectively the results of patients treated with pulmonary hydatid cysts in the past 18 years, considering video-thoracoscopy. Twenty eight patients were treated during this period in 31 cases. Hydatid disease affected only the lung in case of 22 patients, while in 6 cases it was present in the liver and lung simultaneously. Pulmonary hydatid disease affected one side in 24 and both sides in 4 cases. For surgical treatment pericystectomy in one, atypical segment resection in 18, anatomical segmentectomy in three, lobectomy in 7 and video-thoracoscopy in 3 cases were performed without surgical complications. The mean hospital stay was 10.5 days in case of thoracotomies and 8.5 days in case of video-thoracoscopy. There was one recurrence in conventional surgery and reoperation was necessary. After video-thoracoscopy no recurrence was detected. Mean follow-up was 120 months, after video-thoracoscopy it was 20 months. Three patients have uncertain chest pain after thoracotomy, but none has any complaints after video-thoracoscopy. Fifteen patients took mebendazole permanently after the final histological result. According to the authors' practice the indication of lung resections for pulmonary hydatid cysts is limited, in selective cases video-thoracoscopic cystectomy can be a successful treatment of choice.     

Effect of NGF, BDNF, bFGF, aFGF and cell density on NPY expression in cultured rat dorsal root ganglion neurones

The effect of neurotrophic factors on neuropeptide Y (NPY) expression was studied in adult rat dispersed dorsal root ganglion (DRG) cultures. Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), acidic fibroblast growth factor (aFGF) or basic FGF was included in the culture medium during incubation for 72 h. In untreated cultures, around 18% of all neurones (visualized by antibodies to PGP 9.5) expressed NPY-like immunoreactivity (LI). In contrast, in vivo uninjured neurones do not contain detectable levels of NPY-LI. In the immunohistochemical analysis aFGF increased the percentage of NPY-immunoreactive (-IR) neurones 1.8-fold, while NGF, BDNF or bFGF had no significant effect on NPY expression. When the effect of these growth factors was monitored with non-radioactive in situ hybridization, both aFGF and bFGF caused a significant increase (2.25- and 1.8-fold, respectively), whereas, again, NGF and BDNF had no effect. The results also showed an effect of cell density on NPY expression, whereby fewer neurones expressed NPY in high than in low density cultures. This difference was seen in untreated as well as growth factor-treated cultures. The present results support the hypothesis that DRG neurones in culture are in an axotomized state, since they express NPY to about the same extent as axotomized DRG neurones in vivo. Surprisingly, two growth factors of the FGF family enhance NPY expression in DRG neurones, which is in apparent contrast to a published in vivo study [Ji, R.-R., Zhang, Q., Pettersson, R.F., H?kfelt, T., 1996. aFGF, bFGF and NGF differentially regulate neuropeptide expression in dorsal root ganglia after axotomy and induce autotomy. Reg. Pept. 66, 179-189.]. Finally, NPY expression was also influenced by cell density.