bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhanced In vitro and In vivo Cytotoxicity of Combined Reovirus and Radiotherapy
Authors: Twigger Katie , Vidal Laura , White Christine L. , De Bono Johann S. , Bhide Shreerang , Coffey Matt , Thompson Brad , Vile Richard G. , Heinemann Lucy , Pandha Hardev S. , Errington Fiona , Melcher Alan A. , Harrington Kevin J.
Published: February 2008

Purpose: To test combination treatment schedules of reovirus and radiation in human and murine tumor cells in vitro and in vivo.

Experimental Design: In vitro cytotoxicity and cell cycle effects of reovirus given alone and combined with radiotherapy were assessed by colorimetric, tissue culture infectious dose 50, and fluorescence-activated cell sorting–based assays. Interactions between the agents were evaluated using combination index analysis. The effect of different schedules of reovirus and radiotherapy on viral replication and cytotoxicity was tested in vitro and the combination was assessed in three tumor models in vivo.

Results: Characterization of reovirus cytotoxicity in a panel of cell lines yielded a range of sensitivities. Combined reovirus and radiotherapy yielded statistically significantly increased cytotoxicity, particularly in cell lines with moderate susceptibility to reovirus alone. The enhanced cytotoxicity of the combination occurred independently of treatment sequence or schedule. Radiation did not affect viral replication and only reduced reoviral cytotoxicity after clinically irrelevant single doses (>50 Gy). Combination index analysis revealed synergy between radiation (3-10 Gy) and reovirus at multiplicities of infection between 0.001 and 1. Combination treatment significantly increased apoptosis in tumor cells relative to either single-agent treatment. In vivo studies using xenograft and syngeneic tumors showed enhanced activity of the combination relative to reovirus or radiation alone (P < 0.001).

Conclusions: Combining reovirus and radiotherapy synergistically enhances cytotoxicity in a variety of tumor cells in vitro and in vivo. These results offer strong support for translational clinical trials of reovirus plus radiotherapy that have been initiated in the clinic.
Combined Clinical Trial Results of a HER2/neu(E75) Vaccine for the Prevention of Recurrence in High-Risk Breast Cancer Patients: U.S. Military Cancer Institute Clinical Trials Group Study I-01 and I-02
Authors: Peoples George E. , Holmes Jarrod P. , Hueman Matthew T. , Mittendorf Elizabeth A. , Amin Asna , Khoo Steven , Dehqanzada Zia A. , Gurney Jennifer M. , Woll Michael M. , Ryan Gayle B. , Storrer Catherine E. , Craig Dianna , Ioannides Constantin G. , Ponniah Sathibalan
Published: February 2008

Purpose: E75 is an immunogenic peptide from the HER2/neu protein, which is overexpressed in many breast cancer patients. We have conducted two overlapping E75 vaccine trials to prevent recurrence in node-positive (NP) and node-negative (NN) breast cancer patients.

Experimental Design: E75 (HER2/neu 369-377) + granulocyte macrophage colony-stimulating factor was given intradermally to previously treated, disease-free NP breast cancer patients in a dose escalation safety trial and to NN breast cancer patients in a dose optimization study. Local and systemic toxicity was monitored. Immunologic responses were assessed using in vitro assays and in vivo delayed-type hypersensitivity responses. Clinical recurrences were documented.

Results: One hundred and eighty-six patients were enrolled in the two studies (NP, 95; NN, 91). Human leucocyte antigen A2 (HLA-A2) and HLA-A3 patients were vaccinated (n = 101), whereas all others (n = 85) were followed prospectively as controls. Toxicities were minimal, and a dose-dependent immunologic response to the vaccine was shown. Planned primary analysis revealed a recurrence rate of 5.6% in vaccinated patients compared with 14.2% in the controls (P = 0.04) at a median of 20 months follow-up. As vaccine-specific immunity waned over time, the difference in recurrence lost significance at 26 months median follow-up (8.3% versus 14.8%); however, a significant difference in the pattern of recurrence persisted.

Conclusions: E75 is safe and effective in raising a dose-dependent HER2/neu immunity in HLA-A2 and HLA-A3 NP and NN breast cancer patients. More importantly, E75 may reduce recurrences in disease-free, conventionally treated, high-risk breast cancer patients. These findings warrant a prospective, randomized phase III trial of the E75 vaccine with periodic booster to prevent breast cancer recurrences.
Effect of Melanoma on Immune Function in the Regional Lymph Node Basin
Authors: Negin Benjamin , Panka David , Wang Wei , Siddiqui Mustaqueem , Tawa Nicholas , Mullen John , Tahan Steven , Mandato Lucy , Polivy Adam , Mier James , Atkins Michael
Published: February 2008

Purpose: To determine if melanoma within the tumor microenvironment will result in immunosuppression within the draining lymph node as measured by down-regulation of T-cell receptor ζ (TCR ζ) expression.

Experimental Design: Patients with clinical stage I to III melanoma undergoing wide local excision and sentinel lymph node biopsy or therapeutic lymph node dissection were consented to have a portion of their lymph node sampled. Lymph nodes were classified as macroscopically involved (TI), microscopically involved (MI), noninvolved with positive wide excision (NI+), or noninvolved with negative wide excision (NI−). Lymphocytes were stained using antibodies to TCR ζ and other immune cells and analyzed via flow cytometer. Reverse transcription-PCR was used to assess for mediators of immunosuppression.

Results: Fifty patient lymph node samples (15 TI, 7 MI, 9 NI+, and 19 NI−) were evaluated. Increasing involvement of tumor in the lymph node was associated with decreasing TCR ζ chain expression (TI 56%, MI 76%, and NI− 89%). Differences between TI and MI (P = 0.005), TI and NI− (P = 0.0001), and MI and NI− (P = 0.019) were statistically significant. There was also a significant difference between TCR ζ chain expression of NI+ and NI− (73% versus 89%; P = 0.0016). A trend toward increased arginase expression in tumor-involved lymph nodes was detected by reverse transcription-PCR.

Conclusions: Melanoma involvement of regional nodes is associated with loss of TCR ζ expression that is inversely related to tumor burden. Residual melanoma within the wide local excision specimen is associated with TCR ζ loss in noninvolved sentinel lymph nodes, suggesting that immune modulation precedes tumor spread.
A Phase 1 Pharmacokinetic and Pharmacodynamic Study of the Histone Deacetylase Inhibitor Belinostat in Patients with Advanced Solid Tumors
Authors: Steele Nicola L. , Plumb Jane A. , Vidal Laura , Tjørnelund Jette , Knoblauch Poul , Rasmussen Annie , Ooi Chean Eng , Buhl-Jensen Peter , Brown Robert , Evans T.R. Jeffry , DeBono Johann S.
Published: February 2008

Purpose: To determine the safety, dose-limiting toxicity, maximum tolerated dose, and pharmacokinetic and pharmacodynamic profiles of the novel hydroxamate histone deacetylase inhibitor belinostat (previously named PXD101) in patients with advanced refractory solid tumors.

Experimental Design: Sequential dose-escalating cohorts of three to six patients received belinostat administered as a 30-min i.v. infusion on days 1 to 5 of a 21-day cycle. Pharmacokinetic variables were evaluated at all dose levels. Pharmacodynamic measurements included acetylation of histones extracted from peripheral blood mononuclear cells, caspase-dependent cleavage of cytokeratin-18, and interleukin-6 levels.

Results: Forty-six patients received belinostat at one of six dose levels (150-1,200 mg/m2/d). Dose-limiting toxicities were grade 3 fatigue (one patient at 600 mg/m2; one patient at 1,200 mg/m2), grade 3 diarrhea combined with fatigue (one patient at 1,200 mg/m2), grade 3 atrial fibrillation (one patient at 1,200 mg/m2; one patient at 1,000 mg/m2), and grade 2 nausea/vomiting leading to inability to complete a full 5-day cycle (two patients at 1,000 mg/m2). The maximum tolerated dose was 1,000 mg/m2/d. I.v. belinostat displayed linear pharmacokinetics with respect to Cmax and AUC. The intermediate elimination half-life was 0.3 to 1.3 h and was independent of dose. Histone H4 hyperacetylation was observed after each infusion and was sustained for 4 to 24 h in a dose-dependent manner. Increases in interleukin-6 levels were detected following belinostat treatment. Stable disease was observed in a total of 18 (39%) patients, including 15 treated for ≥4 cycles, and this was associated with caspase-dependent cleavage of cytokeratin-18. Of the 24 patients treated at the maximum tolerated dose (1,000 mg/m2/d), 50% achieved stable disease.

Conclusions: I.v. belinostat is well tolerated, exhibits dose-dependent pharmacodynamic effects, and has promising antitumor activity.
T Cell Receptor Genotyping and HOXA/TLX1 Expression Define Three T Lymphoblastic Lymphoma Subsets which Might Affect Clinical Outcome
Authors: Baleydier Frederic , Decouvelaere Anne-Valerie , Bergeron Julie , Gaulard Philippe , Canioni Danielle , Bertrand Yves , Lepretre Stephane , Petit Barbara , Dombret Herve , Beldjord Kheira , Molina Thierry , Asnafi Vahid , Macintyre Elizabeth
Published: February 2008

Purpose: T lymphoblastic lymphomas (T-LBL) are rare disorders of immature T cells which predominantly involve the mediastinum. Their oncogenic pathways and prognostic variables are not clear.

Experimental Design: We undertook a retrospective study of 41 cytoplasmic CD3+ T-LBL (nine cases aged <16 years) by assessing stage of maturation arrest based on T cell receptor (TCR) immunogenotyping, immunohistochemistry, and quantification of the oncogenes thought to be important in immature T cell malignancies.

Results: Application of a TCR-based immunogenetic classification allowed the identification of three subcategories: 11 immature IM0/D-LBL showed no TCR or only incomplete TCRD DJ rearrangement and corresponded to cytoplasmic CD3+ precursors of uncertain lineage. Sixteen mature TCRDdel-LBL showed biallelic TCRD deletion and both TCRG and TCRB rearrangement, consistent with TCRαβ lineage restriction. Fourteen intermediate LBL (Int-LBL) showed complete TCRD VDJ and TCRG VJ rearrangement, with TCRB VDJ rearrangement in the majority. All Int-LBL expressed HOX11/TLX1 or HOXA9 transcripts and a proportion of the latter were associated with CALM-AF10 or NUP214-ABL fusion transcripts. IM0/D-LBL were restricted to adults with extrathymic disease and bone marrow involvement, whereas Int-LBL and TCRDdel-LBL were found in children and adults with predominantly thymic disease. In adults, the Int-LBL subgroup was associated with a significantly superior clinical outcome. This subgroup can be identified either by TCR immunogenotyping or HOXA9/TLX1 transcript quantification.

Conclusion: Application of this molecular classification will allow the prospective evaluation of prognostic effects within pediatric and adult protocols.
An Exploratory Study of Systemic Administration of the Toll-like Receptor-7 Agonist 852A in Patients with Refractory Metastatic Melanoma
Authors: Dummer Reinhard , Hauschild Axel , Becker Juergen C. , Grob Jean-Jacques , Schadendorf Dirk , Tebbs Veronica , Skalsky Jeannine , Kaehler Katharina C. , Moosbauer Stephanie , Clark Ruth , Meng Tze-Chiang , Urosevic Mirjana
Published: February 2008

Purpose: A topical Toll-like receptor 7 (TLR7) agonist induces regression of cutaneous melanocytic neoplasms. We explored antitumor activity of a systemically administered TLR7 agonist, 852A, in patients with metastatic melanoma.

Experimental Design: We undertook a phase II, multicenter, open-label study in patients with chemotherapy-refractory metastatic melanoma. Patients received i.v. 852A, starting at 0.6 mg/m2 and increasing to 0.9 mg/m2 based on tolerance, thrice per week for 12 weeks. Clinical response was determined by Response Evaluation Criteria in Solid Tumors. Immune effects of 852A were monitored by measuring serum type I IFN and IP-10 together with assessment of immune cell markers in peripheral blood.

Results: Twenty-one patients were enrolled. Thirteen patients completed the initial 12-week treatment cycle, with two discontinuing for adverse events considered to be possibly related to study drug. Four (19%) patients had disease stabilization for >100 days. One patient had a partial remission after two treatment cycles, but progressed during the third. Dose-limiting toxicity was observed in two patients. Serum type I IFN and IP-10 increased in most patients on 852A administration. Serum type I IFN increases were greater after dosing with 852A 0.9 mg/m2 than after 0.6 mg/m2 (P = 0.009). The maximal increase in IP-10 compared with baseline correlated with the maximal increase in type I IFN (P = 0.003). In the eight patients with immune cell marker data, CD86 expression on monocytes increased significantly post-first dose (P = 0.007).

Conclusion: Intravenous 852A was well tolerated and induced systemic immune activation that eventually resulted in prolonged disease stabilization in some patients with stage IV metastatic melanoma who had failed chemotherapy.
Imaging Epidermal Growth Factor Receptor Expression In vivo: Pharmacokinetic and Biodistribution Characterization of a Bioconjugated Quantum Dot Nanoprobe
Authors: Diagaradjane Parmeswaran , Orenstein-Cardona Jacobo M. , E. Colón-Casasnovas Norman , Deorukhkar Amit , Shentu Shujun , Kuno Norihito , Schwartz David L. , Gelovani Juri G. , Krishnan Sunil
Published: February 2008

Purpose: To develop and validate an optical imaging nanoprobe for the discrimination of epidermal growth factor (EGF) receptor (EGFR)–overexpressing tumors from surrounding normal tissues that also expresses EGFR.

Experimental Design: Near-infrared (NIR) quantum dots (QD) were coupled to EGF using thiol-maleimide conjugation to create EGF-QD nanoprobes. In vitro binding affinity of these nanoprobes and unconjugated QDs was evaluated in a panel of cell lines, with and without anti-EGFR antibody pretreatment. Serial optical imaging of HCT116 xenograft tumors was done after systemic injection of QD and EGF-QD.

Results: EGF-QD showed EGFR-specific binding in vitro. In vivo imaging showed three distinct phases, tumor influx (∼3 min), clearance (∼60 min), and accumulation (1-6 h), of EGF-QD nanoprobes. Both QD and EGF-QD showed comparable nonspecific rapid tumor influx and clearance followed by attainment of an apparent dynamic equilibrium at ∼60 min. Subsequently (1-6 h), whereas QD concentration gradually decreased in tumors, EGF-QDs progressively accumulated in tumors. On delayed imaging at 24 h, tumor fluorescence decreased to near-baseline levels for both QD and EGF-QD. Ex vivo whole-organ fluorescence, tissue homogenate fluorescence, and confocal microscopic analyses confirmed tumor-specific accumulation of EGF-QD at 4 h. Immunofluorescence images showed diffuse colocalization of EGF-QD fluorescence within EGFR-expressing tumor parenchyma compared with patchy perivascular sequestration of QD.

Conclusion: These results represent the first pharmacokinetic characterization of a robust EGFR imaging nanoprobe. The measurable contrast enhancement of tumors 4 h after systemic administration of EGF-QD and its subsequent normalization at 24 h imply that this nanoprobe may permit quantifiable and repetitive imaging of EGFR expression.
The Effect of Vitamin E on Hot Flashes in Menopausal Women
Authors: Ziaei S. , Kazemnejad A. , Zareai M.
Published: February 2008

Background: Hot flashes affect as many as 75% of menopausal women. Estrogen reliably reduces the severity of hot flashes and remain the single most effective treatment. Today, however, more and more women are seeking alternatives. Instead of hormonal therapy, women are turning to vitamins, and other over-the-counter products for relief from hot flashes. This study was undertaken to assess the effect of vitamin E on hot flashes. Method: A placebo double blind-controlled trial was conducted. After 1 week baseline period, the enrolled patients (n = 51) received placebo (identical in appearance to vitamin E softgel) daily for 4 weeks, followed by 1 week wash out and 400 IU vitamin E (softgel cap) daily for the next 4 weeks. Diary was used to measure hot flashes before and at the end of the study. Result: There were statistical significant differences in hot flashes severity score (2.37 ± 0.74, 1.80 ± 0.87) and their daily frequency (5.00 ± 3.34, 3.19 ± 2.74) after the treatments between the placebo and vitamin E therapies (p < 0.0001). Conclusion: Based on our trial, vitamin E is recommended for the treatment of hot flashes.
Mild-to-Moderate Active Luminal Crohn’s Disease
Authors: Michetti Pierre , Juillerat Pascal , Mottet Christian , Pittet Valérie , Gonvers Jean-Jacques , Vader John-Paul , Froehlich Florian , Felley Christian
Published: February 2008

The management of luminal Crohn’s disease, the most common form of initial presentation of the disease, depends on the location and the severity of the disease. Mild-to-moderate disease represents a relatively large proportion of patients with a first flare of luminal disease, which may also be associated with perianal disease. As quality of life of these patients inversely correlates with disease activity, adequate therapy is a central goal of the overall patient management. Treatment options include mainly sulfasalazine, budesonide and systemic steroids, while the role of mesalazine and antibiotics remains controversial. The role of biological therapies has not been thoroughly evaluated in patients with mild disease.
Severe and Steroid-Resistant Crohn’s Disease
Authors: Michetti Pierre , Mottet Christian , Juillerat Pascal , Pittet Valérie , Felley Christian , Vader John-Paul , Gonvers Jean-Jacques , Froehlich Florian
Published: February 2008

Patients with moderate-to-severe disease and patients with steroid-refractory or steroid-dependent disease differ in their management, as the latter two groups usually involve patients whose condition is less acute. Systemic corticosteroids represent the mainstay of the management of moderate-to-severe disease and remain the first-line therapy in this setting. Anti-TNF agents represent choice alternatives for patients who do not respond to steroids or in whom steroids are contraindicated. Purine analogues, methotrexate and infliximab have all shown efficacy in achieving steroid-free remission in patients with steroid-refractory or steroid-dependent disease. Other fast-acting immunosuppressors showed little benefit. Surgery may be indicated in this setting. Natalizumab may prove useful in patients refractory to infliximab and other anti-TNF agents.
Fistulizing Crohn’s Disease
Authors: Felley Christian , Mottet Christian , Juillerat Pascal , Pittet Valérie , Froehlich Florian , Vader John-Paul , Michetti Pierre , Gonvers Jean-Jacques
Published: February 2008

Fistulas are common in Crohn’s disease. A population-based study has shown a cumulative risk of 33% after 10 years and 50% after 20 years. Perianal fistulas were the most common (54%). Medical therapy is the main option for perianal fistula once abscesses, if present, have been drained, and should include antibiotics (both ciprofloxacin and metronidazole) and immunomodulators. Infliximab should be reserved for refractory patients. Surgery is often necessary for internal fistulas.
Fibrostenotic Crohn’s Disease
Authors: Froehlich Florian , Juillerat Pascal , Mottet Christian , Pittet Valérie , Felley Christian , Vader John-Paul , Gonvers Jean-Jacques , Michetti Pierre
Published: February 2008

Crohn’s disease is often complicated by gastrointestinal strictures. Postoperative recurrence at the anastomotic site is common and repeated surgical interventions may be necessary. Medical treatment may relieve active inflammation (see chapter on active luminal disease) but fibrous strictures will not respond to this. Mechanical treatment methods consist of endoscopic balloon dilation, stricturoplasty or surgical resection. Fibrostenotic Crohn’s disease does not respond to medical therapy and requires endoscopic or surgical treatment.
Maintenance of Surgically Induced Remission of Crohn’s Disease
Authors: Froehlich Florian , Juillerat Pascal , Pittet Valérie , Felley Christian , Mottet Christian , Vader John-Paul , Michetti Pierre , Gonvers Jean-Jacques
Published: February 2008

At 1 year after a first resection, up to 80% of patients show an endoscopic recurrence, 10–20% have clinical relapse, and 5% have surgical recurrence. Smoking is one of the most important risk factors for postoperative recurrence. Preoperative disease activity and the severity of endoscopic lesions in the neoterminal ileum within the first postoperative year are predictors of symptomatic recurrence. Mesalazine is generally the first-line treatment used in the postoperative setting but still provokes considerable controversy as to its efficacy, in spite of the results of a meta-analysis. Immunosuppressive treatment (azathioprine, 6-mercaptopurine) is based on scant evidence but is currently used as a second-line treatment in postsurgical patients at high risk for recurrence, with symptoms or with early endoscopic lesions in the neoterminal ileum. Nitroimidazole antibiotics (metronidazole, ornidazole) are also effective in the control of active Crohn’s disease in the postoperative setting. Given their known toxicity, they may be used as a third-line treatment as initial short-term prevention therapy rather than in the long term. Conventional corticosteroids, budesonide or probiotics have no proven role in postoperative prophylaxis. Infliximab has not as yet been studied for use in the prevention of relapse after surgery.
Pregnancy and Breastfeeding in Patients with Crohn’s Disease
Authors: Mottet Christian , Juillerat Pascal , Pittet Valérie , Gonvers Jean-Jacques , Froehlich Florian , Vader John-Paul , Michetti Pierre , Felley Christian
Published: February 2008

Crohn’s disease commonly affects women of childbearing age. Available data on Crohn’s disease and pregnancy show that women with Crohn’s disease can expect to conceive successfully, carry to term and deliver a healthy baby. Control of disease activity before conception and during pregnancy is critical, to optimize both maternal and fetal health. Generally speaking, pharmacological therapy for Crohn’s disease during pregnancy is similar to pharmacological therapy for nonpregnant patients. Patients maintained in remission by way of pharmacological therapy should continue it throughout their pregnancy. Sulfasalazine, mesalazine and corticosteroids are safe, azathioprine and 6-mercaptopurine are reasonably safe with few discordant data, infliximab seems safe as well, whereas methotrexate is contraindicated during pregnancy. During breastfeeding, mesalazine and prednisone are considered safe, azathioprine/6-mercaptopurine, budesonide and infliximab probably safe and methotrexate is contraindicated.
Drug Safety in Crohn’s Disease Therapy
Authors: Juillerat Pascal , Pittet Valérie , Felley Christian , Mottet Christian , Froehlich Florian , Vader John-Paul , Gonvers Jean-Jacques , Michetti Pierre
Published: February 2008

The management of Crohn’s disease usually consists of a succession of short-term acute phase treatments followed by a long-term maintenance therapy. Above all the most frequent adverse events and the data on the long-term safety of the therapeutic arsenal available to the physician will be taken into consideration. The drugs described in this article include 5-ASA compounds, antibiotics (metronidazole, ciprofloxacin and rifaximin), corticosteroids (budesonide, prednisone and equivalents), thiopurines (azathioprine and 6-mercaptopurine), methotrexate, anti-tumor necrosis factor inhibitors (infliximab, adalimumab, certolizumab), natalizumab, anticalcineurin inhibitors (cyclosporine, tacrolimus) and mycophenolate mofetil.
Evidence for a Viral Neuropathy in Recurrent Vertigo
Authors: Gacek Richard R.
Published: February 2008

The concept that reactivation of latent neurotropic viruses (i.e. Herpesviridae group) in the vestibular ganglion is responsible for recurrent vestibulopathies is presented. A similar histopathologic degeneration of vestibular ganglion cells in vestibular neuronitis (VN), Ménière’s disease and benign paroxysmal positional vertigo is presented to support this concept. The clinical response (relief of vertigo) to the administration of antiviral medication in these syndromes provides practical evidence of a viral neuropathy in patients with recurrent vertigo. Relief of vertigo after this treatment was 90% in VN, Ménière’s disease and VN. The relief of positional vertigo (benign paroxysmal positional vertigo) was 66%.
Does the Serological Study for Viral Infection in Autoimmune Inner Ear Disease Make Sense?
Authors: García-Berrocal José Ramón , Ramírez-Camacho Rafael , González-García José Angel , Verdaguer José María , Trinidad Almudena
Published: February 2008

Viral infections of the labyrinth have been considered a major source of auditory and vestibular system pathology. However, the involvement of virus in the development of immune reactions responsible for immunomediated inner ear disease has not been studied enough. Following viral infection, an effector immune response, humoral (B cell) and/or cytotoxic (T cell) is directed against a virus and it might cross-react with self-protein or autoantigen, evoking an autoimmune response. Since clinically it can be very difficult to establish a viral etiology for such disorders, serologic studies can be used to confirm the suspected diagnosis. Patients affected by immunomediated inner ear disease that had presented an upper respiratory tract infection underwent an immunologic workup study including microbiological study. After the application of this diagnostic protocol, only one patient, that was subsequently diagnosed with Cogan’s syndrome, showed a positive serological test for viral infection. On the basis of the low efficacy of serological testing and due to the lack of evidence, we do not recommend to carry out serologic studies for viral infection.
Intratympanic Application of an Antiviral Agent for the Treatment of Ménière’s Disease
Authors: Guyot Jean-Philippe , Maire Raphael , Delaspre Olga
Published: February 2008

It has been suggested that Ménière’s disease is part of a polyganglionitis in which symptoms result from the reactivation of neurotropic virus within the internal auditory canal, and that intratympanic applications of an antiviral agent might be an efficient therapy. In 2002, we performed a pilot study ending with encouraging results. Control of vertigo was achieved in 80% of the 17 patients included. We present here a prospective, double-blind study, with a 2-year follow-up, in 29 patients referred by ENT practitioners for a surgical treatment after failure of a medical therapy. The participation in the study was offered to patients prior to surgery. A solution of ganciclovir 50 mg/ml or of NaCl 9% was delivered for 10 consecutive days via a microwick inserted into the tympanic membrane in the direction of the round window or through a ventilation tube. One patient was withdrawn from the study immediately after the end of the injections. He could not complete the follow-up period, because of persisting vertigo. As he had received the placebo, he was then treated with the solution of ganciclovir. Symptoms persisted and he underwent a vestibular neurectomy. Among the remaining 28 patients, surgery could be postponed in 22 (81%). Surgery remained necessary to control vertigo in 3 patients from the group that received the antiviral agent, and in 3 from the control group. Using an analogical scale, patients of both groups indicated a similar improvement of their health immediately after the intratympanic injections. The scores obtained with a 36-item short-form health survey quality of life questionnaire and the Dizziness Handicap Inventory were also similar for both groups. In conclusion, most patients were improved after the intratympanic injections, but there was no obvious difference between the treated and control groups. The benefit might be due to the middle ear ventilation or reflect an improvement in the patients’ emotional state.
Do Viruses Cause Inner Ear Disturbances?
Authors: Pyykkö I. , Zou J.
Published: February 2008

The association of viral infection to inner ear disease is controversial. Experiments on animals show that several viruses are capable of causing hearing loss, if applied into the perilymph. Some of these have specific affinity to the cellular type of the inner ear, as sensory epithelia and cochlear nerve. Some viruses as adenoviruses and Coxsackie virus B have specific CAR receptors that are identified in different cell types, whereas other act by attaching onto nonspecific cellular surface receptors. Some viruses such as varicella zoster virus (VZV) do not cause disease in rodents. We assessed 273 patients with clinical, serological, neuro-otologic and endoscopic evaluations. Of the 273 patients, 43 served as control subjects. The patients either had Ménière’s disease (n = 158), recurrent vertigo of unknown etiology (n = 56), or hearing loss (n = 17). Antibodies against neurotropic and common viruses were evaluated. VZV, influenza B, CBV5 and RSV titers were significantly elevated in patients with inner ear disease when compared with the control group. In analyzing the internal relationship, VZV and influenza B were intercorrelated. We did not find a correlation between hearing loss and viral titers. In conclusion, VZV, Coxsackie virus B5 and influenza B virus may be the main causes of inner ear disorder. The spiral and Scarpa’s ganglion are potential sites harboring viral DNA for possible latent infection.
Viral Etiology for Inner Ear Diseases: Proven, Unproven, Unlikely
Authors: Oliveira Carlos A. , Sampaio André L. , Bahmad, Jr. Fayez M. , Araújo Mercedes F.S.
Published: February 2008

This is a revision article that deals with the broad field of inner ear disease caused by viral infections. Some of these entities have been proven to have a viral etiology. Others have strong evidence in favor of a viral causation but still cannot be considered as a viral disease. Finally, other entities are suggestive of a viral etiology but when the whole body of evidence is considered one concludes that a viral etiology is indeed unlikely. We review the literature and add our own experience in this subject. Clearly, the most important evidence about this subject came from the study of temporal bone histopathology. Certainly, we can learn much more if we continue to collect and study temporal bone specimens histopathologically.
Specific transcriptional changes in human fetuses with autosomal trisomies
Authors: Altug-Teber Ö. , Bonin M. , Walter M. , Mau-Holzmann U.A. , Dufke A. , Stappert H. , Tekesin I. , Heilbronner H. , Nieselt K. , Riess O.
Published: February 2008

Among full autosomal trisomies, only trisomies of chromosome 21 (Down syndrome), 18 (Edwards syndrome) and 13 (Patau syndrome) are compatible with postnatal survival. But the mechanisms, how a supernumerary chromosome disrupts the normal development and causes specific phenotypes, are still not fully explained. As an alternative to gene dosage effect due to the trisomic chromosome a genome-wide transcriptional dysregulation has been postulated. The aim of this study was to define the transcriptional changes in trisomy 13, 18, and 21 during early fetal development in order to obtain more insights into the molecular etiopathology of aneuploidy. Using oligonucleotide microarrays, we analyzed whole genome expression profiles in cultured amniocytes (AC) and chorionic villus cells (CV) from pregnancies with a normal karyotype and with trisomies of human chromosomes 13, 18 and 21. We observed a low to moderate up-regulation for a subset of genes of the trisomic chromosomes. Transcriptional levels of most of the genes on the supernumerary chromosome appeared similar to the respective chromosomal pair in normal karyotypes. A subset of chromosome 21 genes including the DSCR1 gene involved in fetal heart development was consistently up-regulated in different prenatal tissues (AC, CV) of trisomy 21 fetuses whereas only minor changes were found for genes of all other chromosomes. In contrast, in trisomy 18 vigorous downstream transcriptional changes were found. Global transcriptome analysis for autosomal trisomies 13, 18, and 21 supported a combination of the two major hypotheses.
Intragenic breakpoints localized by array CGH in a t(2;6) familial translocation
Authors: Bernheim A. , Toujani S. , Guillaud-Bataille M. , Richon C. , Waxin H. , Dessen P. , Berger R.
Published: February 2008

A 244K genome-wide array based comparative genomic hybridization study was carried out in a familial translocation t(2;6)(p25;p21) balanced in the mother and unbalanced in her daughter. In the past, this translocation has allowed us to localize the HLA multigene cluster to chromosome 6. With microarray technology, confirmation of the chromosome localization of the HLA system was easily obtained, showing that such approach may be applied to the breakpoint localizations of other familial structural changes when they are unbalanced. The disruption of genes at the translocation breakpoints that did not have any phenotypic consequences in the parent will allow the generation of a map of ‘haplotolerant genes’. In addition, many genomic variants were detected with this technology, enlarging the possibility of analyzing their possible contribution to phenotypic diversity.
Cytogenetic sensitivity of G0 lymphocytes of Fanconi anemia patients and obligate carriers to mitomycin C and ionizing radiation
Authors: Mohseni-Meybodi A. , Mozdarani H. , Vosough P.
Published: February 2008

It is well known that Fanconi anemia (FA) patients show a hypersensitivity to the effect of cross-linking agents such as mitomycin C (MMC) and diepoxybutane (DEB), while the sensitivity of these patients to ionizing radiation is still controversial. Fanconi anemia heterozygotes do not show a hypersensitivity to the above mentioned agents compared to normal individuals. To examine the radio-sensitivity of Fanconi anemia patients and heterozygotes, ten patients and 13 heterozygotes were enrolled in this study. Standard metaphase analysis for detection and verification of radio-sensitivity was used to establish the relationship between γ-ray and chromosome breakages in these groups. Statistical analysis was used for the assessment of aberrations including chromatid and chromosome breaks and exchanges. Results of chromosome aberration yield that: (i) differentiation between obligate carriers and the control group after MMC treatment and gamma irradiation was not possible; (ii) homozygotes were clearly distinguishable from heterozygotes and controls after MMC treatment; (iii) FA patients don’t show hypersensitivity to gamma irradiation compared to normal controls and heterozygous carriers.
U-HO1, a new cell line derived from a primary refractory classical Hodgkin lymphoma
Authors: Mader A. , Brüderlein S. , Wegener S. , Melzner I. , Popov S. , Müller-Hermelink H.K. , Barth T.F. , Viardot A. , Möller P.
Published: February 2008

The Hodgkin cell line U-HO1 was established from a malignant pleural effusion of a 23-year-old male patient during the end stage of refractory nodular sclerosing classical Hodgkin lymphoma (cHL). Since its establishment in 2005, U-HO1 has maintained stable characteristics in vitro and has a doubling time of about 4 days under standard culture conditions. U-HO1 forms typical Reed-Sternberg cells in suspension, is EBV negative, lacks HLA-A, -B, -C but expresses HLA-D proteins/CD74 and exposes CD15 together with CD30 in the absence of CD19 and CD20 on the cell surface. Karyotype analysis of U-HO1 revealed a hyperdiploid karyotype with multiple clonal aberrations. Most significant is an elongated chromosome 2, der(2)t(2;10)(q35; q16.1)add(2)(p13). CGH analysis revealed the following imbalances: ish cgh dim(1)(p13p31)(p12q21), enh(2)(p13p23), dim(4)(q31.3qter), enh(6)(q22q27), enh(12), enh(18), enh(20) (q13.1pter). FISH analysis showed about six-fold amplification of REL and BCL11A, thus, U-HO1 is prototypical for cHL in every aspect tested so far. As an outstanding feature compared to the existing HL cell lines, U-HO1 has high levels of microRNA transcripts of MIRN216 and MIRN217 located in the amplicon 2p16. Compared to other HL cell lines, U-HO1 proved far less genetically aberrant suggesting that U-HO1’s imbalances suffice to cause the full-blown phenotype of primary refractory cHL.
Chromosomal localization of the telomeric (TTAGGG)n sequence in eight species of New World Primates (Neotropical Primates, Platyrrhini)
Authors: Mudry M.D. , Nieves M. , Bolzán A.D.
Published: February 2008

Chromosomal localization of the telomeric sequence (TTAGGG)n in eight New World Primates (Platyrrhini) (Alouatta caraya, Alouatta palliata, Alouatta guariba clamitans, Aotus azarae, Ateles chamek, Cebus nigritus, Cebus paraguayanus, and Saimiri boliviensis) using Fluorescence In Situ Hybridization (FISH) with a peptide nucleic acid (PNA) pantelomeric probe and their possible relationship with the C-banding pattern were analyzed. FISH showed telomeric signals only at the terminal regions of chromosomes from all the species analyzed. Although all of them showed centromeric C+ bands and different size and location of extracentromeric C+ bands, none, except Aotus azarae exhibited (peri)centromeric interstitial telomere-like sequences (ITS). The presence of ITS in Aotus azarae was limited to one pair of submetacentric chromosomes and very likely represents telomeric sequences remaining after a fusion event of ancestral chromosomes during karyotype evolution. Therefore, our data indicate that the distribution of heterochromatin blocks do not correlate with the presence of ITS. However, we cannot rule out the possibility that simple ITS arrays with a few copies of the (TTAGGG)n sequence, not detectable by conventional FISH, might play a role in the karyotypic evolution of Ceboidea. Further FISH and molecular studies will be needed to confirm this hypothesis.
Characterization of a balanced reciprocal translocation, rcp(9;11)(q27;q11) in cattle
Authors: De Lorenzi L. , De Giovanni A. , Molteni L. , Denis C. , Eggen A. , Parma P.
Published: February 2008

Cytogenetic analysis of a phenotypically normal young bull from Marchigiana breed revealed the presence of an abnormal karyotype. The observation of longer and smaller chromosomes than BTA1 and BTA29, respectively in all metaphases suggested the presence of a reciprocal translocation. RBG-banding confirmed this hypothesis revealing the involvement of BTA9 and BTA11. FISH analyses using cattle-specific BAC clones (474A12 and 293G09 for BTA9; 035D03 for BTA11) identified rcp(9;11)(q27;q11) in the two regions affected. Moreover analyses performed on both parents established the ‘de novo’ origin of the anomaly. Comparison with human homologue sequences (HSA6q24.3→q25.3 for BTA9q27 and HSA2q11.1→q12.1 for BTA11q11) revealed that both breakpoint regions are gene rich as up to date at least 200 genes have been localized in these regions. Thus, further analyses are required to identify the sequences disrupted by the breakpoints and to verify their consequences on rcp carrier phenotype.
Chromosome number reduction accompanied by extensive heterochromatin addition in the bat Glauconycteris beatrix (Mammalia; Chiroptera, Vespertilionidae)
Authors: Volleth M. , Heller K.G.
Published: February 2008

The presumed ancestral karyotype of the bat family Vespertilionidae consists of 44 chromosomes with a fundamental number of autosomal arms (FNa) of 50. Previously, only two of the roughly 350 vespertilionid species have been reported with 2n lower than 26. In this paper we report the 2n = 22 karyotype of the African vespertilionid Glauconycteris beatrix which shows an X-autosome translocation and extended, paracentromeric, chromomycin-A3-positive heterochromatin.
High chromosomal polymorphism and heterozygosity in Cyclocephala tridentata from Guadeloupe: chromosome comparison with some other species of Dynastinae (Coleoptera: Scarabaeidae)
Authors: Dutrillaux A.M. , Xie H. , Dutrillaux B.
Published: February 2008

Very distinct karyotypes have been observed in two Cyclocephala species from Guadeloupe, considered as very close and possibly vicariant: C. insulicola with only metacentric and C. tridentata tridentata with many acrocentric autosomes. This prompted us to study the karyotype of a few other neotropical Dynastinae belonging to four of the eight existing tribes, to find out which one of these two species had the most divergent chromosomes from their ancestral condition. In the four additional species studied, i.e., Cyclocephalamaffafa, Strategus syphax, Ligyrus cuniculus and Megasoma actaeon, a karyotype composed of 20 chromosomes, including 18 meta- or submetacentric autosomes was found, as it was in C. insulicola. Thus, the karyotype of C. t. tridentata, in which most of the 18 autosomes were acrocentric, is apomorphic. In addition, it was highly polymorphic, with six different karyotypes observed among the ten specimens studied. All had one to four heterozygous chromosome pairs formed by one acrocentric and one submetacentric carrying a large juxta-centromeric heterochromatic component. This heterozygosity did not seem to impair either meiotic synapsis or chiasma formation and chromosome segregation. Such high rates of chromosome heterozygosity and polymorphism are infrequent and never described in beetles. This suggests that C. t. tridentata undergoes an active process of chromosome evolution. A possible relationship with insularity and/or pesticide exposure is briefly discussed.
Comparison of mealybug (Planococcus lilacinus) and fruit fly genomes: isolation and analysis of conserved sequences and their utility in studying synteny in the mealybug
Authors: Mohan K.N. , Rani B.S. , Selvam S. , Debarshi S. , Kadandale J.S.
Published: February 2008

By using ligation-mediated PCR products from mealybug DNA as tester and biotinylated fly DNA as driver, we recovered a fraction of the tester that remains hybridized to driver following high-stringency washing conditions. This fraction is expected to contain mealybug sequences conserved in the fly (MCF). Reciprocal experiments enabled the isolation of fly sequences conserved in the mealybug (FCM). Coding sequences among MCF show amino acid identities >40% with fly proteins, allowing a reliable identification of orthologs. Three sequences from the fly cytogenetic positions 98–99 were hybridized onto mealybug chromosomes and the results identified differences in synteny between the two species. Taken together, our results present a method for direct isolation of sequences conserved between an ‘orphan’ (mealybug) genome and a ‘reference’ (fly) genome and showed that these sequences can be used to study chromosome synteny in the mealybug.
Condensation of rye chromatin in somatic interphase nuclei of Ph1 and ph1b wheat
Authors: Kopecky D. , Allen D.C. , Duchoslav M. , Dolezel J. , Lukaszewski A.J.
Published: February 2008

The Ph1 locus in hexaploid wheat (Triticum aestivum L.) enforces diploid-like behavior in the first metaphase of meiosis. To test the hypothesis that this chromosome pairing control is exercised by affecting the degree of chromatin condensation, the dispersion of rye chromatin in interphase nuclei in somatic tissues of wheat-rye chromosome translocations 1RS.1BL, 2RS.2BL, 2BS.2RL, 3RS.3DL and 5RS.5BL was compared in Ph1 and ph1b isogenic backgrounds. No significant differences in rye chromatin condensation that could be attributed to the Ph1 locus were detected. Regardless of the Ph1 status, each rye chromosome arm tested conformed to the general Rabl’s orientation and occupied portions of the nuclei proportional to their length. Earlier observations that indicated the involvement of Ph1 locus in rye chromatin condensation in wheat could have been due either to specific loci on the studied 5RL rye arm that control the chromosome condensation process or to damage to the genetic system controlling chromatin condensation in the existing ph1b stocks of wheat. That damage might have been caused by homoeologous recombination and uneven disjunction of chromosomes from multivalents.
Isolation and characterization of the highly repeated fraction of the banana genome
Authors: Hřibová E. , Doleželová M. , Town C.D. , Macas J. , Doležel J.
Published: February 2008

Although the nuclear genome of banana (Musa spp.) is relatively small (1C ∼ 610 Mbp for M. acuminata), the results obtained from other sequenced genomes suggest that more than half of the banana genome may be composed of repetitive and non-coding DNA sequences. Knowledge of repetitive DNA can facilitate mapping of important traits, phylogenetic studies, BAC-based physical mapping, and genome sequencing/annotation. However, only a few repetitive DNA sequences have been characterized in banana. In this work, we used DNA reassociation kinetics to isolate the highly repeated fraction of the banana genome (M. acuminata ‘Calcutta 4’). Two libraries, one prepared from Cot ≤0.05 DNA (2,688 clones) and one from Cot ≤0.1 sequences (4,608 clones), were constructed, and 614 DNA clones were chosen randomly for sequencing and further characterization. Dot-plot analysis revealed that 14% of the sequenced clones contained various semi-tandem and palindromic repeated sequences. ‘BLAST’ homology searches showed that, in addition to tandem repeats, the Cot libraries were composed mainly of different types of retrotransposons, the most frequent being the Ty3/gypsy type monkey retrotransposon. Selected sequences displaying tandem organization properties were mapped by PRimed IN Situ DNA labeling (PRINS) to the secondary constriction on metaphase chromosomes of M. acuminata ‘Calcutta 4’. Southern hybridization with selected BAC clones carrying 45S rDNA confirmed the presence of the tandem repeats in the 45S rDNA unit. This work significantly expands the knowledge of the repetitive fraction of the Musa genome and organization of its chromosomes.
Identification and characterization of a new type of asymmetrical dicentric chromosome derived from a single maternal chromosome 18
Authors: Lin C.C. , Li Y.-C. , Liu P.-P. , Hsieh L.-J. , Cheng Y.-M. , Teng R.-H. , Shi S.-L. , Tsai F.-J.
Published: February 2008

Molecular cytogenetic analysis identified a new type of dicentric chromosome involving different breakpoints at 18q in a female fetus. The chromosome anomaly was designated as an asymmetrical pseudoisodicentric chromosome 18, 46,XX,psu dic(18)(pter→q11.2::q21.3→pter)mat. A series of BAC clones for 18q11.2 and q21.3 regions were used to identify one breakpoint within the region q11.2 between 19.8 and 21.6 Mb from the telomere of 18p and another breakpoint within q21.3 between 55.4 and 56.9 Mb from the telomere of 18p by FISH analysis. Real-time quantitative PCR and microsatellite analysis further verified that the dicentric chromosome was maternal in origin and resulted from a break-reunion between sister chromatids of a single maternal chromosome. We propose that a loop-type configuration of sister chromatids took place and that the break-reunion occurred at cross sites of the loop to form an asymmetrical isodicentric chromosome during either mitosis or meiosis. In this case, the asymmetrical pseudoisodicentric resulted in an 18pter→ q11.2 duplication and an 18q21.3→qter deletion, which could have led to certain dysmorphic features of 18q– syndrome in this fetus.