The isolation of differentially expressed cDNA clones from the filarial nematode <i>Brugia pahangi</i>

A cDNA library constructed from 3 day post-infective L3 of the filarial nematode Brugia pahangi was screened by differential hybridization with cDNA probes prepared from different life-cycle stages. Five cDNA clones hybridizing selectively to the mosquito-derived L3 probe were isolated and characterized. Northern blot analysis of 4 of the clones confirmed that each was most highly expressed in the mosquito-derived L3. The expression of each mRNA during parasite development in the mosquito vector was investigated using RT-PCR, and all were shown to be abundant in the immature L3. Four of the 5 cDNAs cloned coded for structural proteins: 2 cuticular collagens, and the muscle proteins tropomyosin and troponin. Further studies on troponin using an antiserum raised to the recombinant protein demonstrated that the protein, unlike the mRNA, was present in all life-cycle stages examined, while immunogold labelling demonstrated that it was localized to the muscle blocks

Dynamic instability in resonant tunneling

We show that an instability may be present in resonant tunneling through a quantum well in one, two and three dimensions, when the resonance lies near the emitter Fermi level. A simple semiclassical model which simulates the resonance and the projected density of states by a nonlinear conductor, the Coulomb barrier by a capacitance, and the time evolution by an iterated map, is used. The model reproduces the observed hysteresis in such devices, and exhibits a series of bifurcations leading to fast chaotic current fluctuations.Comment: 7 pages, 2 figure

Reply Comment: Comparison of Approaches to Classical Signature Change

We contrast the two approaches to ``classical" signature change used by Hayward with the one used by us (Hellaby and Dray). There is (as yet) no rigorous derivation of appropriate distributional field equations. Hayward's distributional approach is based on a postulated modified form of the field equations. We make an alternative postulate. We point out an important difference between two possible philosophies of signature change --- ours is strictly classical, while Hayward's Lagrangian approach adopts what amounts to an imaginary proper ``time" on one side of the signature change, as is explicitly done in quantum cosmology. We also explain why we chose to use the Darmois-Israel type junction conditions, rather than the Lichnerowicz type junction conditions favoured by Hayward. We show that the difference in results is entirely explained by the difference in philosophy (imaginary versus real Euclidean ``time"), and not by the difference in approach to junction conditions (Lichnerowicz with specific coordinates versus Darmois with general coordinates).Comment: 10 pages, latex, no figures. Replying to - "Comment on `Failure of Standard Conservation Laws at a Classical Change of Signature'", S.A. Hayward, Phys. Rev. D52, 7331-7332 (1995) (gr-qc/9606045

Bright solitary-matter-wave collisions in a harmonic trap: Regimes of solitonlike behavior

Systems of solitary waves in the one-dimensional Gross-Pitaevskii equation, which models a trapped atomic Bose-Einstein condensate, are investigated theoretically. To analyze the soliton nature of these solitary waves, a particle analogy for the solitary waves is formulated. Exact soliton solutions exist in the absence of an external trapping potential, which behave in a particlelike manner, and we find the particle analogy we employ to be a good model also when a harmonic trapping potential is present up to a gradual shift in the trajectories when the harmonic trap period is short compared with the collision time of the solitons. We find that the collision time of the solitons is dependent on the relative phase of the solitons as they collide. In the case of two solitons, the particle model is integrable, and the dynamics are completely regular. In the case of a system of two solitary waves of equal norm, the solitons are shown to retain their phase difference for repeated collisions. This phase preservation can be used to find regimes where there is agreement between the wave and particle models. This also implies that soliton regimes may be found in three-dimensional geometries where solitary waves can be made to repeatedly collide out of phase, stabilizing the condensate against collapse. The extension to three particles supports both regular and chaotic regimes. The trajectory shift observed for two solitons carries over to the case of three solitons. This shift aside, the agreement between the particle model and the wave dynamics remains good, even in chaotic regimes

Bright matter-wave soliton collisions in a harmonic trap : regular and chaotic dynamics

Collisions between bright solitary waves in the 1D Gross-Pitaevskii equation with a harmonic potential, which models a trapped atomic Bose-Einstein condensate, are investigated theoretically. A particle analogy for the solitary waves is formulated and shown to be integrable for a two-particle system. The extension to three particles is shown to support chaotic regimes. Good agreement is found between the particle model and simulations of the full wave dynamics, suggesting that the dynamics can be described in terms of solitons both in regular and chaotic regimes, presenting a paradigm for chaos in wave mechanics

Beauty is only mucosa deep : a retrospective analysis of oral lumps and bumps caused by cosmetic fillers

Introduction: The injection of dermal fillers into orofacial tissues is becoming increasingly popular for cosmetic purposes, in particular for lip augmentation. Both natural and synthetic filler materials are available, producing a spectrum of clinical and histological appearances. Aims: The aim of this study was to review the clinicopathological characteristics of dermal filler cases from 2006 to 2016, reported at a specialist oral pathology unit. Methods: An archival search of the pathology database was performed to retrieve cases reported as being consistent with cosmetic fillers. Results: Ten cases of orofacial cosmetic fillers were retrieved. Of these cases, 100% were from female patients and the mean age of presentation was 47.6 years (range 24-68 years). The lips were the most frequently involved site (80%, n = 8). The majority of provisional diagnoses were related to salivary gland disease, including neoplasms (30%, n = 3), cysts (20%, n = 2) or inflammatory disease (10%, n = 1). Only two cases (20%) were clinically thought to be related to previous cosmetic injections. A variety of filler materials were seen, including collagen, hydroxyapatite and silicone. However, hyaluronic acid-based materials were the most common (50%, n = 5). Conclusions: Complications of cosmetic dermal fillers are becoming more common and should be considered within a differential diagnosis for unusual orofacial swellings

Diethylstilbestrol induces oxidative DNA damage, resulting in apoptosis of spermatogonial stem cells in vitro

YesThe spermatogonial stem cells (SSCs) are the only germline stem cells in adults that are responsible for the transmission of genetic information from mammals to the next generation. SSCs play a very important role in the maintenance of progression of spermatogenesis and help provide an understanding of the reproductive biology of future gametes and a strategy for diagnosis and treatment of infertility and male reproductive toxicity. Androgens/oestrogens are very important for the suitable maintenance of male germ cells. There is also evidence confirming the damaging effects of oestrogen-like compounds on male reproductive health. We investigated the effects in vitro, of diethylstilbestrol (DES) on mouse spermatogonial stem cells separated using Staput unit-gravity velocity sedimentation, evaluating any DNA damage using the Comet assay and apoptotic cells in the TUNEL assay. Immunocytochemistry assays showed that the purity of isolated mouse spermatogonial cells was 90%, and the viability of these isolated cells was over 96%. Intracellular superoxide anion production (O2−) in SSCs was detected using p-Nitro Blue Tetrazolium (NBT) assay. The viability of cells after DES treatment was examined in the CCK8 (cell counting kit-8) cytotoxicity assay. The results showed that DES-induced DNA damage causes an increase in intracellular superoxide anions which are reduced by the flavonoid, quercetin. Investigating the molecular mechanisms and biology of SSCs provides a better understanding of spermatogonial stem cell regulation in the testis

In vitro responses to known in vivo genotoxic agents in mouse germ cells

YesGenotoxic compounds have induced DNA damage in male germ cells and have been associated with adverse clinical outcomes including enhanced risks for maternal, paternal and offspring health. DNA strand breaks represent a great threat to the genomic integrity of germ cells. Such integrity is essential to maintain spermatogenesis and prevent reproduction failure. The Comet assay results revealed that the incubation of isolated germ cells with n-ethyl-n-nitrosourea (ENU), 6-mercaptopurine (6-MP) and methyl methanesulphonate (MMS) led to increase in length of Olive tail moment and % tail DNA when compared with the untreated control cells and these effects were concentration-dependent. All compounds were significantly genotoxic in cultured germ cells. Exposure of isolated germ cells to ENU produced the highest concentration-related increase in both DNA damage and gene expression changes in spermatogonia. Spermatocytes were most sensitive to 6-MP, with DNA damage and gene expression changes while spermatids were particularly susceptible to MMS. Real-time PCR results showed that the mRNA level expression of p53 increased and bcl-2 decreased significantly with the increasing ENU, 6-MP and MMS concentrations in spermatogonia, spermatocytes and spermatids respectively for 24 hr. Both are gene targets for DNA damage response and apoptosis. These observations may help explain the cell alterations caused by ENU, 6-MP and MMS in spermatogonia, spermatocytes and spermatids. Taken together, ENU, 6-MP and MMS induced DNA damage and decreased apoptosis associated gene expression in the germ cells in vitro.Libyan Governmen

Germ Cell Responses to Doxorubicin Exposure in Vitro

YesAnthracyclines such as doxorubicin (Dox), widely used to treat various types of tumours, may result in induced testicular toxicity and oxidative stress. The present investigation was designed to determine whether exposure of isolated and purified mouse germ cells to Dox induces DNA damage in the form of strand breaks (presumably) resulting in apoptosis and to investigate the relative sensitivity of specific cell types. DNA damage was assessed using the Comet assay and the presence of apoptosis was determined by TUNEL assay. Isolated mouse germ cells were treated with different concentrations (0.05, 0.5 and 1 mM, respectively) of Dox, and fixed 1 h after treatment. The incidences of both DNA damage shown by single cell gel-electrophoresis and of apoptosis increased significantly in each specific cell type in a concentration-dependent manner. The DNA damage and apoptosis incidences gradually increased with concentration from 0.05 to 1 mM with Dox. Our results indicate that apoptosis plays a vital role in the induction of germ cell phase-specific toxicity caused by Dox with pre-meiotically and meiotically dividing spermatogonia and spermatocytes respectively as highly susceptible target cells.Higher Education Funding Council for England (HEFCE

Silver nanoparticle-mediated cellular responses in isolated primary Sertoli cells in vitro

YesThe present study explored the mechanism of cytotoxic and genotoxic effects of AgNPs on a primary culture of mouse Sertoli cells in vitro. To understand the possible molecular mechanisms of testicular lesions following exposure to AgNPs, isolated Sertoli cells were exposed to 5, 10, or 15 μg/ml. DNA damage in the Comet assay and apoptosis in the TUNEL assay were evaluated. The mRNA expression of p53 and bcl-2 genes and their proteins involved in apoptosis was also investigated. The antioxidant status of treated Sertoli cells was determined by measuring superoxide dismutase (SOD-1), catalase (CAT), and glutathione peroxidase (GPX-1) and superoxide dismutase (SOD-1) using quantitative polymerase chain reaction (qPCR)qPCR. The superoxide anions were detected using the nitroblue tetrazolium (NBT) reduction assay. Results indicated that AgNP exposure causes increased oxidative stress levels. The activation of p53, repression of bcl-2 and reduction of endogenous antioxidant enzymes were also involved in these mechanistic pathways, leading to reduced cell numbers and cell detachment.The Sponsorship of the Libyan Government of a PhD studentship to Khaled Haba