Selective deletion of cochlear hair cells causes rapid age-dependent changes in spiral ganglion and cochlear nucleus neurons

During nervous system development, critical periods are usually defined as early periods during which manipulations dramatically change neuronal structure or function, whereas the same manipulations in mature animals have little or no effect on the same property. Neurons in the ventral cochlear nucleus (CN) are dependent on excitatory afferent input for survival during a critical period of development. Cochlear removal in young mammals and birds results in rapid death of target neurons in the CN. Cochlear removal in older animals results in little or no neuron death. However, the extent to which hair-cell-specific afferent activity prevents neuronal death in the neonatal brain is unknown. We further explore this phenomenon using a new mouse model that allows temporal control of cochlear hair cell deletion. Hair cells express the human diphtheria toxin (DT) receptor behind the Pou4f3 promoter. Injections of DT resulted in nearly complete loss of organ of Corti hair cells within 1 week of injection regardless of the age of injection. Injection of DT did not influence surrounding supporting cells directly in the sensory epithelium or spiral ganglion neurons (SGNs). Loss of hair cells in neonates resulted in rapid and profound neuronal loss in the ventral CN, but not when hair cells were eliminated at a more mature age. In addition, normal survival of SGNs was dependent on hair cell integrity early in development and less so in mature animals. This defines a previously undocumented critical period for SGN survival

Ultrasound-guided topographic anatomy of the medial calcaneal branches of the tibial nerve

Background: The purpose of this study was to evaluate the topographic anatomy of the tibial nerve and its medial calcaneal branches in relation to the tip of the medial malleolus and to the posterior superior tip of the calcaneal tuberosity using the ultrasound examination and to verify its preoperative usefulness in surgical treatment. Materials and methods: Bilateral ultrasound examination was performed on 30 volunteers and the location of the tibial nerve bifurcation and medial calcaneal branches origin were measured. Medial calcaneal branches were analysed in reference to the amount and their respective nerves of origin. Results: In 77% of cases, tibial nerve bifurcation occurred below the tip of the medial malleolus with the average distance of 5.9 mm and in 48% of cases above the posterior superior tip of the calcaneal tuberosity with the average distance of 2.7 mm. In 73% of cases medial calcaneal branches occurred as a single branch originating from the tibial nerve (60%). The average distance of the first, second and third medial calcaneal branch was accordingly 9.3 mm above, 9.5 mm below and 11.6 mm below the tip of the medial malleolus and 17.7 mm above, 1.6 mm below and 4 mm below the posterior superior tip of the calcaneal tuberosity. Conclusions: As the tibial nerve and its branches present a huge variability in the medial ankle area, in order to prevent the iatrogenic injuries, the preoperative or intraoperative ultrasound assessment (sonosurgery) of its localisation should be introduced into the clinic

Calcium confusion--is the variability in calcium response by Sertoli cells to specific hormones meaningful or simply redundant?

When results of more than ten different studies on hormone-induced calcium signals in Sertoli cells are taken together, a wide variety of responses emerges. The reported changes range from increased concentrations, via no response at all, to decreased calcium concentrations. Minor variations in cell isolation techniques, culture conditions, or techniques for measuring the intracellular calcium could explain some of these differences. However, erratic variations in response are also observed within research groups under very similar experimental conditions. Such 'negative' findings are mainly reported orally and do not further penetrate the scientific community. As hormone-dependent calcium responses evidently may depend very much on the context of the cells, calcium transients would appear to be unreliable bioassay principles with which to detect the primary actions of FSH and effectors such as androgens on Sertoli cells. A more important biological question is whether these sometimes opposed calcium transients are connected with a particular cellular response. To date there is no evidence for such a tight coupling in Sertoli cells, implying that, at least under in vitro conditions, calcium signals might even be redundant altogether. Such calcium variability is probably not unique to Sertoli cells, and the aim of this commentary is to promote an open debate that may help to transform the current state of 'calcium confusion' into a better understanding of the intracellular calcium language

40 Zakażenie wirusem Epsteina-Barr w ziarnicy złośliwej. Ocena korelacji między ekspresją LMP-1 w komórkach Hodgkina i Reed-Sternberga, a ekspresją BCL-2

Wynik badań epidemiologicznych i immunologicznych wskazują na zbieżność między zachorowaniem na ziarnicę złośliwą a zakażeniem wirusem Epsteina-Barr (EBV). Wykrycie wirusa EBV w morfologicznie złośliwych komórkach ziarniczych sugeruje, że wirus wniknął do komórki przed transformacją nowotworową i klonalną proliferacją, zwłaszcza, że wyłącznie 1 na 10 limfocytów jest zainfekowany wirusem.Ekspresji LMP-1 białku błonowemu wirusa przypisuje się rolę karcynogenną Białko to poprzez indukcję protoonkogenu Bcl-2 może zapobiegać śmierci komórki w mechanizmie apoptozy. Analizowano materiał pochodzący od 43 chorych na ziarnicę złośliwą w różnym stopniu zaawansowania klinicznego oraz histopatologicznego. W oparciu o metody immunohistochemiczne oceniano ekspresję Lmp-1 i Bcl-2w komórkach Reed-Sternberga i Hodkina. Otrzymane wyniki mogą potwierdzać powyższą tezę

Large Scale Gene Expression Profiles of Regenerating Inner Ear Sensory Epithelia

Loss of inner ear sensory hair cells (HC) is a leading cause of human hearing loss and balance disorders. Unlike mammals, many lower vertebrates can regenerate these cells. We used cross-species microarrays to examine this process in the avian inner ear. Specifically, changes in expression of over 1700 transcription factor (TF) genes were investigated in hair cells of auditory and vestibular organs following treatment with two different damaging agents and regeneration in vitro. Multiple components of seven distinct known signaling pathways were clearly identifiable: TGFβ, PAX, NOTCH, WNT, NFKappaB, INSULIN/IGF1 and AP1. Numerous components of apoptotic and cell cycle control pathways were differentially expressed, including p27KIP and TFs that regulate its expression. A comparison of expression trends across tissues and treatments revealed identical patterns of expression that occurred at identical times during regenerative proliferation. Network analysis of the patterns of gene expression in this large dataset also revealed the additional presence of many components (and possible network interactions) of estrogen receptor signaling, circadian rhythm genes and parts of the polycomb complex (among others). Equal numbers of differentially expressed genes were identified that have not yet been placed into any known pathway. Specific time points and tissues also exhibited interesting differences: For example, 45 zinc finger genes were specifically up-regulated at later stages of cochlear regeneration. These results are the first of their kind and should provide the starting point for more detailed investigations of the role of these many pathways in HC recovery, and for a description of their possible interactions

Cosmic ray tests of the D0 preshower detector

The D0 preshower detector consists of scintillator strips with embedded wavelength-shifting fibers, and a readout using Visible Light Photon Counters. The response to minimum ionizing particles has been tested with cosmic ray muons. We report results on the gain calibration and light-yield distributions. The spatial resolution is investigated taking into account the light sharing between strips, the effects of multiple scattering and various systematic uncertainties. The detection efficiency and noise contamination are also investigated.Comment: 27 pages, 24 figures, submitted to NIM

ADAM10 and gamma-secretase regulate sensory regeneration in the avian vestibular organs

The loss of sensory hair cells from the inner ear is a leading cause of hearing and balance disorders. The mammalian ear has a very limited ability to replace lost hair cells, but the inner ears of non-mammalian vertebrates can spontaneously regenerate hair cells after injury. Prior studies have shown that replacement hair cells are derived from epithelial supporting cells and that the differentiation of new hair cells is regulated by the Notch signaling pathway. The present study examined molecular influences on regeneration in the avian utricle, which has a particularly robust regenerative ability. Chicken utricles were placed in organotypic culture and hair cells were lesioned by application of the ototoxic antibiotic streptomycin. Cultures were then allowed to regenerate in vitro for seven days. Some specimens were treated with small molecule inhibitors of γ-secretase or ADAM10, proteases which are essential for transmission of Notch signaling. As expected, treatment with both inhibitors led to increased numbers of replacement hair cells. However, we also found that inhibition of both proteases resulted in increased regenerative proliferation. Subsequent experiments showed that inhibition of γ-secretase or ADAM10 could also trigger proliferation in undamaged utricles. To better understand these phenomena, we used RNA-Seq profiling to characterize changes in gene expression following γ-secretase inhibition. We observed expression patterns that were consistent with Notch pathway inhibition, but we also found that the utricular sensory epithelium contains numerous γ-secretase substrates that might regulate cell cycle entry and possibly supporting cell-to-hair cell conversion. Together, our data suggest multiple roles for γ-secretase and ADAM10 in vestibular hair cell regeneration

Genetic engineering and molecular characterization of yeast strain expressing hybrid human-yeast squalene synthase as a tool for anti-cholesterol drug assessment

AIMS: The main objective of the study is molecular and biological characterization of the human-yeast hybrid squalene synthase (SQS), as a promising target for treatment of hypercholesterolaemia. METHODS AND RESULTS: The human-yeast hybrid SQS, with 67% amino acids, including the catalytic site derived from human enzyme, was expressed in Saccharomyces cerevisiae strain deleted of its own SQS gene. The constructed strain has a decreased level of sterols compared to the control strain. The mevalonate pathway and sterol biosynthesis genes are induced and the level of triacylglycerols is increased. Treatment of the strain with rosuvastatin or zaragozic acid, two mevalonate pathway inhibitors, decreased the amounts of squalene, lanosterol and ergosterol, and up-regulated expression of several genes encoding enzymes responsible for biosynthesis of ergosterol precursors. Conversely, expression of the majority genes implicated in the biosynthesis of other mevalonate pathway end products, ubiquinone and dolichol, was down-regulated. CONCLUSIONS: The S. cerevisiae strain constructed in this study enables to investigate the physiological and molecular effects of inhibitors on cell functioning. SIGNIFICANCE AND IMPACT OF THE STUDY: The yeast strain expressing hybrid SQS with the catalytic core of human enzyme is a convenient tool for efficient screening for novel inhibitors of cholesterol-lowering properties