The role of RNase 7 in innate cutaneous defense against Pseudomonas aeruginosa

The ribonuclease RNase 7 is a major skin‐derived human antimicrobial protein expressed in keratinocytes. Here we show that the gram‐negative pathogen Pseudomonas aeruginosa secretes factor(s) that induced RNase 7 gene and protein expression in human primary keratinocytes. The metalloprotease inhibitor marimastat, the epidermal growth factor receptor (EGFR) inhibitor AG1478 and the EGFR blocking antibody cetuximab significantly attenuated this induction, indicating an important role of the EGFR for the P. aeruginosa‐mediated RNase 7 induction. In line with this, siRNA‐mediated downregulation of ADAM17, a metalloprotease known to proteolytically mediate the release of soluble EGFR ligands, decreased the P. aeruginosa‐mediated RNase 7 induction in keratinocytes. The impact of the EGFR was also demonstrated in a human 3D skin equivalent where blockade of the EGFR diminished induction of RNase 7 by P. aeruginosa. Blockade of Toll‐like receptor 5 (TLR5), a pattern recognition receptor (PRR) known to be activated by P. aeruginosa, only moderately reduced the P. aeruginosa‐mediated RNase 7 induction in keratinocytes. The functional relevance of RNase 7 to participate in cutaneous defense against P. aeruginosa was demonstrated by antibodies that neutralized the antimicrobial activity of RNase 7. These antibodies significantly inhibited the capacity of human stratum corneum skin extracts to control growth of P. aeruginosa. Taken together, our results indicate that P. aeruginosa induces the expression of RNase 7 in keratinocytes in an EGFR‐dependent manner. Enhanced release of RNase 7 contributes to control cutaneous growth of P. aeruginosa.     

Communication in medical education: Students' Demands

Medicine, Health Care and Philosophy -     

Phytoestrogens role in bone functional structure protection in the ovariectomized rat

Effects of soy protein diet on bone formation and density were evaluated in ovariectomized rats as a model for postmenopausal women. Twenty-seven 9-month-old rats were assigned to 3 treatment groups for the 9-week study: sham-surgery (Sh, n = 9); ovariectomy (Ovx, n = 9); ovariectomy + soy diet (OvxS, n = 9). Rats had free access to an AIN-93 M diet or AIN-93 M diet with 7% soy protein concentration and water. At sacrifice, rear legs were removed, and the right femur and tibia were cleaned manually. Serum alkaline phosphatase, a marker of bone formation, was measured colorimetrically. Bone density was measured using Archimedes' Principle. Alkaline phosphatase activity was greater in OvxS (114 +/- 19 U/L) and Ovx (128 +/- 26 U/L) compared to Sh (110 +/- 22 U/L). Femur bone density was greater for OvxS (1.520 +/- 0.02 g/cc) compared to Ovx (1.510 +/- 0.017 g/cc), but not to Sh (1532 +/- 0.025 g/cc). Tibia bone density was greater for OvxS (1.560 +/- 0.019 g/cc) compared to Ovx (1.553 +/- 0.015 g/cc), but not to Sh (1566 +/- 0.03 g/cc). In conclusion soy protein diet increased the rate of bone formation and bone density in some bones, suggesting that may help prevent bone loss in postmenopausal women.     

Long-term potentiation as a substrate for memory: evidence from studies of amygdaloid plasticity and Pavlovian fear conditioning

Recent reports have raised concerns about the ability of long-term potentiation (LTP) to account for associative learning and memory. In this paper, we review the many mechanistic similarities between one form of associative learning, Pavlovian fear conditioning, and amygdaloid LTP. We then address many of the criticisms levied against LTP within the framework of fear conditioning. We believe that many of the apparent discrepancies between LTP and behavior can be generally accounted for by a failure to appreciate that learned behavior is supported by multiple synapses in an extensive network of brain structures. We conclude that LTP remains a viable substrate for memory.     

Effects of locally administered pentylenetetrazole on nigral single unit activity and severity of dystonia in a genetic model of paroxysmal dystonia

The dt(sz) hamster is a well-established animal model of idiopathic paroxysmal dystonia. Previous investigations of this mutant have indicated dysfunctions of the gamma-aminobutyric acid (GABA)-ergic system within the basal ganglia. Systemic administration of the central stimulant pentylenetetrazole (PTZ) aggravated dystonia at subconvulsant doses, whereas GABA-mimetic drugs have beneficial effects in dt(sz) hamsters. GABA mimetics also provide clinical benefit in humans with idiopathic paroxysmal dystonia. The spontaneous discharge rates of substantia nigra pars reticulata (SNr) neurons was unaltered in anesthetized dt(sz) hamsters, but systemic application of subconvulsant doses of PTZ caused significantly greater increases of discharge rates in dystonic hamsters compared with nondystonic controls. The present study tested the hypothesis that SNr neurons are more sensitive to local application of PTZ in dt(sz) hamsters than in nondystonic hamsters. PTZ applied locally by pressure injection at 2, 3, and 5 mM to the SNr during in vivo single unit recordings revealed a dose-dependent increase of SNr discharge rates in mutants and controls relative to predrug rates, with a significantly greater increase in mutants at 3 mM PTZ. To examine the functional relevance of the increased susceptibility of SNr neurons to PTZ in mutants, the effects of PTZ on severity of dystonia were investigated after microinjections into the SNr of freely moving dt(sz) hamsters. Bilateral nigral microinjection of 40 ng PTZ did not aggravate dystonia but exerted moderate antidystonic effects. Therefore, the previous findings of prodystonic effects of systemic administration of PTZ in dt(sz) hamsters are related to extranigral effects rather than to the elevation of nigral discharge rates in response to systemic, or locally applied, PTZ. The greater susceptibility of neurons within the SNr to PTZ suggests dysfunctions of the GABA(A) receptor in dt(sz) mutants.     

Primary dystonia: Is abnormal functional brain architecture linked to genotype?

The DYT1 dystonia mutation is associated with an abnormal metabolic brain network characterized by hypermetabolism of the basal ganglia, supplementary motor area, and the cerebellum. In this study, we quantified the activity of this network in carriers of other dystonia mutations to determine whether this functional abnormality is linked to genotype. The findings suggest that the DYT1 metabolic topography is not genotype specific and may be present in carriers of other dystonia mutations.     

Multicolor-FISH analysis of a natural killer cell line (NK-92)

Relatively little is known about the cytogenetics of natural killer (NK) neoplasms, and the emergence of recurrent structural alterations involving specific chromosomal breakpoints is still in its infancy. This gap has doubtless hampered identification of the oncogene alterations posited to underly NK tumors. We describe in detail the cytogenetic rearrangements present in a cytotoxic NK cell line (NK-92) established from a patient with large granular lymphocyte (LGL) lymphoma. The NK-92 cell line is one of very few cytotoxic NK cell lines described and the first to be used clinically. Cytogenetic analysis was performed independently using two multicolor-fluorescence in situ hybridization (M-FISH) systems, the first M-FISH study of a cell line derived from a NK neoplasm. Several non-random cytogenetic features previously reported in NK cells were, thus, identified, including rearrangements of chromosomes 7 and 17, along with breakpoints at 11q23, 12q12 and 8p22/23. FISH revealed that NK-92 cells carry multiple rearrangements with distinct breakpoints at 8p resembling those previously described in NK lymphoma. Our data strengthen the claim of NK-92 to model NK neoplasms and highlight this cell line as a potential resource for mining relevant oncogenic changes therein.     

Steady-state free precession projection MRI as a potential alternative to the conventional chest X-ray in pediatric patients with suspected pneumonia

Magnetic resonance imaging of the lung tissue is thought to be hardly possible due to physical limitations especially the low proton density, susceptibility, and motion artifacts. The objective of our study was to evaluate and refine a very fast MR technique at a low field strength which overcomes the limitations in MR lung imaging. Thirty-five investigations were performed in 30 pediatric patients with suspected pneumonia. The MR investigations were performed in coronal slice orientation without cardiac or respiratory triggering in a low-field MR system. An optimized true fast imaging with steady precession sequence was applied. The MR images and the corresponding conventional chest radiographs were evaluated. The examination time per slice was 1.6 s. No motion artifacts could be observed. The signal-to-noise ratio for pulmonary parenchyma ranged from 4.9 to 7.1. All pathological findings of the chest X-ray images were correctly identified by the MRI (κ=0.82–0.85). Effusions as well as small pneumonic infiltrates were more precisely detected by the MRI investigation (κ=0.82) as compared with X-ray. Low-field projection MRI is a promising alternative to pediatric chest X-ray. Due to its short examination time, it overcomes the physical limits of usual MRI methods and provides comparable diagnostic information. Electronic Publication