Gender biased neuroprotective effect of Transferrin Receptor 2 deletion in multiple models of Parkinson’s disease

Alterations in the metabolism of iron and its accumulation in the substantia nigra pars compacta accompany the pathogenesis of Parkinson’s disease (PD). Changes in iron homeostasis also occur during aging, which constitutes a PD major risk factor. As such, mitigation of iron overload via chelation strategies has been considered a plausible disease modifying approach. Iron chelation, however, is imperfect because of general undesired side effects and lack of specificity; more effective approaches would rely on targeting distinctive pathways responsible for iron overload in brain regions relevant to PD and, in particular, the substantia nigra. We have previously demonstrated that the Transferrin/Transferrin Receptor 2 (TfR2) iron import mechanism functions in nigral dopaminergic neurons, is perturbed in PD models and patients, and therefore constitutes a potential therapeutic target to halt iron accumulation. To validate this hypothesis, we generated mice with targeted deletion of TfR2 in dopaminergic neurons. In these animals, we modeled PD with multiple approaches, based either on neurotoxin exposure or alpha-synuclein proteotoxic mechanisms. We found that TfR2 deletion can provide neuroprotection against dopaminergic degeneration, and against PD- and aging-related iron overload. The effects, however, were significantly more pronounced in females rather than in males. Our data indicate that the TfR2 iron import pathway represents an amenable strategy to hamper PD progression. Data also suggest, however, that therapeutic strategies targeting TfR2 should consider a potential sexual dimorphism in neuroprotective response.Subject terms: Metals, Ageing, Neurological disorders     

Evaluation of the xenobiotic biotransformation capability of six rodent hepatoma cell lines in comparison with rat hepatocytes

Summary Phase I and II activities were examined in six rodent hepatoma cell lines and compared with those of cultured rat hepatocytes both in basal conditions and after exposure to 5μM methylcholanthrene, 2 mM phenobarbital, and 15μMβ-naphtoflavone. The metabolic profile of testosterone was also studied. The highest aryl hydrocarbon hydroxylase and 7-ethoxycoumarinO-deethylase activities were found in MH1C1 cells. Comparable values for 7-ethoxyresorufinO-deethylase activity, ranging from 21.6 to 42.9 pmol/mg × min, were observed in the hepatocytes and hepatoma cells, except the HTC cells. In contrast, only Fao cells showed 7-pentoxyresorufinO-depentylase activity at levels similar to those of hepatocytes (6.2±1.0 and 7.4±1.2 pmol/mg × min, respectively). Rat hepatocytes actively hydroxylatedp-nitrophenol, but this activity was not measurable in hepatoma cells. Glutathione transferase activity was maintained in all the hepatoma cell lines at similar levels to those found in hepatocytes (684 ± 56 nmol/mg × min). The seven hydroxylated metabolites of testosterone produced by cultured hepatocytes were negligible in hepatoma cells. Exposure of cells to inducers revealed that aryl hydrocarbon hydroxylase activity was mainly increased after treatment with 3-methylcholanthrene andβ-naphtoflavone, and the highest values were found in rat hepatocytes followed by MH1C1 and Fao cells. 3-Methylcholanthrene and naphtoflavone treatment also resulted in a marked increase in 7-ethoxyresorufinO-deethylase activity in hepatocytes as well as in H4IIC3, McA-Rh7777, MH1C1, and Fao cells. An enhancement of 7-ethoxycoumarinO-deethylase activity due to the three inducers was observed in both rat hepatocytes and hepatoma cells, with MH1C1 cells treated with methylcholanthrene showing the highest activity (727±74 pmol/mg × min). Increases in 7-pentoxyresorufinO-depentylase activity were detected after phenobarbital treatment of hepatocytes, MH1C1, and Fao cells, whereas a low response was observed in the other hepatoma cells. Of the six hepatoma cell lines examined, MH1C1 and Fao cells are the ones that are most similar to cultured rat hepatocytes in their expression of biotransformation activities.     

Partial trisomy 16q resulting from maternal translocation

Summary A 3 1/2-year-old male with partial trisomy of the long arm of chromosome 16 resulting from a maternal balanced translocation is described. Karyotype: 46,XY,-22,der(22),t(16;22)(q21;p12)mat.     

Overload and neovascularization of shoulder tendons in volleyball players

ABSTRACT: BACKGROUND: In overhead sports like volleyball, the onset of a rotator cuff tendinopathy due to functional overload is a common observation. An angiofibroblastic etiopathogenesis has been hypothesized, whereby a greater anaerobic metabolism occurs in critical zones of the tendon with a lower degree of vascularization; this would induce collagen and extracellular matrix degradation, that could then trigger a compensatory neovascularization response. We performed a clinical observational study of 80 elite volleyball players, monitoring the perfusion values of the rotator cuff tendons by oximetry. RESULTS: No statistically significant differences were found between the oximetry data and age, sex or years of sports activity, nor when comparing the right and left arm or the dominant and non-dominant arm. A statistically significant difference was found for the dominant arm values in relation to the competitive role, higher values being obtained in outside hitters (62.7%) middle hitters (53.7%), opposite hitters (55.5%) and libero players (54.4%) (p<0.05), whereas there was no difference in players with the role of setter (56.2%) (p>0.05). CONCLUSIONS: The different tendon vascularization values found in players with different roles in the team may be attributed to a response to the specific biomechanical demands posed by the different overhead throwing roles.     

Reconstruction of ancestral sequences by the inferential method,a tool for protein engineering studies

This paper describes the inferential method, an approach for reconstructing protein and nucleotide sequences of ancestral species, starting from known, homologous, contemporary sequences. The method requires knowledge of the topology of the phylogenetic tree, whose nodes are the species to whom the reconstructed sequences belong.The method has been tested by computer simulation of speciation and nucleotide substitutions, starting from a single ancestral sequence, and by subsequent reconstruction of nodal sequences. Results have shown that reconstructions obtained by the inferential method are affected by limited error frequencies, which (1) are proportional to the squares of nucleotide substitution rates and of internodal distances, and (2) are little influenced by non-uniformity of transformation rates of nucleotides.Furthermore, good agreement of the results has been obtained by comparing protein-sequence reconstructions carried out with the inferential method with those obtained using the maximum parsimony method in two different cases: e.g., a reconstruction of simulated sequences and a reconstruction of mammalian ribonuclease sequences.Abbreviations used MP maximum parsimony method - ML maximum likelihood method - IM inferential method - MY millions of years - N-tree natural-like phylogenetic tree - E-tree equibranched phylogenetic tree - EA percentage number of erroneous amino acids in a reconstructed sequence - EC percentage number of erroneous codons in a reconstructed sequence - t _n time interval between a P- and its - F-sequence nucleotides and amino acids are indicated by their I.U.B. codes (N.C.-I.U.B., 1985) Correspondence to: A. Di Donato     

Analysis of somatic and salivary gland antigens of third stage larvae of Rhinoestrus spp. (Diptera, Oestridae)

Larvae of Rhinoestrus spp. (Diptera, Oestridae) infect nasal and sinus cavities of horses, causing a nasal myiasis characterized by severe respiratory distress. Presently, the diagnosis of horse nasal botfly relies on the observation of clinical signs, on the post mortem retrieval of larvae or on molecular assays performed using pharyngeal swabs. The present study was carried out to characterize larval somatic proteins and salivary glands of Rhinoestrus spp. in a preliminary assessment towards the immunodiagnosis of equine rhinoestrosis. Out of the 212 necropsied horses 13 were positive for the presence of Rhinoestrus spp. larvae. The analysis of the sera from the infected animals by Western blotting assay showed the presence of a specific host humoral immune response against Rhinoestrus spp. larvae and proved that the salivary glands are the major immunogens in horse nasal botflies.     

Effects of pertussis toxin and indomethacin on murine lymphocytes in the bronchoalveolar lavage fluids

The total number of lymphocytes in the bronchoalveolar lavage (BAL) fluids significantly increased in mice injected intravenously with pertussis toxin (PT), while the absolute number of alveolar macrophages markedly decreased. This finding probably reflects the lymphocyte accumulation in interstitial spaces as we previously observed in mice injected with PT. In addition, indomethacin, at lower dosage (0.5 mg/kg) prevented peripheral lymphocytosis and lymphocyte accumulation in the alveolar spaces of the lungs of mice injected with PT. These results provide evidence that PT is responsible for lymphocyte accumulation together with a marked decrease of alveolar macrophages in the lungs of treated mice; moreover, indomethacin is effective in preventing bronchoalveolar changes caused by PT.