Genomewide copy number alteration screening of circulating plasma DNA: potential for the detection of incipient tumors.

Background: Early cancer diagnosis might improve survival rates. As circulating tumor DNA (ctDNA) carries cancer-specific modifications, it has great potential as a noninvasive biomarker for detection of incipient tumors. Patients and methods: We collected cell-free DNA (cfDNA) samples of 1002 elderly without a prior malignancy, carried out whole-genome massive parallel sequencing and scrutinized the mapped sequences for the presence of (sub)chromosomal copy number alterations (CNAs) predictive for a malignancy. When imbalances were detected, 6-monthly clinical follow-up was carried out. Results: In 3% of participants chromosomal imbalances were detected. Follow-up analyses, including whole-body MRI screening, confirmed the presence of five hematologic malignancies: one Hodgkin lymphoma (HL), stage II; three non-HL (type chronic lymphocytic leukemia, Rai I-Binet A; type SLL, stage III; type mucosa-associated lymphoid tissue, stage I) and one myelodysplastic syndrome with excess blasts, stage II. The CNAs detected in cfDNA were tumor-specific. Furthermore, one case was identified with monoclonal B-cell lymphocytosis, a potential precursor of B-cell malignancy. In 24 additional individuals, CNAs were identified but no cancer diagnosis was made. For 9 of them, the aberrant cfDNA profile originated from peripheral blood cells. For 15 others the origin of aberrations in cfDNA remains undetermined. Conclusion(s): Genomewide profiling of cfDNA in apparently healthy individuals enables the detection of incipient hematologic malignancies as well as clonal mosaicism with unknown clinical significance. CNA screening of cellular DNA of peripheral blood in elderly has established that clonal mosaicism for these chromosomal anomalies predicts a 5- to 10-fold enhanced risk of a subsequent cancer. We demonstrate that cfDNA screening detects CNAs, which are not only derived from peripheral blood, but even more from other tissues. Since the clinical relevance of clonal mosaics in other tissues remains unknown, long-term follow-up is warranted. Taken together, this study demonstrates that genomewide cfDNA analysis has potential as an unbiased screening approach for hematological malignancies and premalignant conditions.

Quantification of trabecular spatial orientation from low-resolution images.

No accepted methodology exists to assess trabecular bone orientation from clinical CT scans. The aim of this study was to test the hypothesis that the distribution of grey values in clinical CT images is related to the underlying trabecular architecture and that this distribution can be used to identify the principal directions and local anisotropy of trabecular bone. Fourteen trabecular bone samples were extracted from high-resolution (30 µm) micro-CT scans of seven human femoral heads. Trabecular orientations and local anisotropy were calculated using grey-level deviation (GLD), a novel method providing a measure of the three-dimensional distribution of image grey values. This was repeated for different image resolutions down to 300 µm and for volumes of interest (VOIs) ranging from 1 to 7 mm. Outcomes were compared with the principal mechanical directions and with mean intercept length (MIL) as calculated for the segmented 30-µm images. For the 30-µm images, GLD predicted the mechanical principal directions equally well as MIL. For the 300-µm images, which are resolutions that can be obtained in vivo using clinical CT, only a small increase (3°-6°) in the deviation from the mechanical orientations was found. VOIs of 5 mm resulted in a robust quantification of the orientation. We conclude that GLD can quantify structural bone parameters from low-resolution CT images.

[Urinary incontinence in the elderly: distinctive feature and management]. / Incontinence du sujet âgé: particularités et prise en charge.

Urinary incontinence is very common among the elderly and results in a series of personal and economic complications. This population is often very heterogeneous with the aetiologies or dependences which are not very obvious. It is thus important to carry out among these patients a total assessment which will make it possible to detect the affections or the dependence from which it suffers. The BGMST (Belgian Geriatric Minimum Screening Test) includes a series of non aggressive tests which will answer this need. The performance of urodynamic tests is not easily achieved in case of not very autonomous or intellectually little collaborating patients. The prescription of surgical treatments is not necessarily to reject (urge incontinence). With regard to drugs, a good knowledge of their metabolism, of their interferences with other treatments in progress, is important especially in the absence of literature taking into account this heterogeneous old population. The prescriptions must be regularly re-examined in order to avoid weakening of these patients. In addition, in the elderly, aetiologies of incontinence are often multiple and consequently its daily management becomes particularly difficult.

Recovery of humoral immunity is critical for successful antiviral therapy in disseminated mouse adenovirus type 1 infection.

Severe adenovirus infections in transplant recipients undergoing immunosuppressive therapy are of increasing concern. Controversy exists on the contribution of antiviral therapy and the host immune response to recovery from these infections. Here, we established a systemic mouse adenovirus type 1 (MAV-1) infection in cyclophosphamide (CyP)-treated BALB/c mice. CyP was administered at 100 mg per kg of body weight every other day for 2, 3, or 4 weeks, thereby inducing general but reversible leukopenia, with a major suppression of the B-cell numbers and functionality that was more pronounced than that seen with T cells. The outcome of MAV-1 infection was dependent on the duration of CyP therapy, as the mice with the most severe immunosuppression were the most vulnerable to MAV-1-induced hemorrhagic enteritis and mortality. The protective effect of concomitant antiviral therapy with cidofovir depended on the level of immunosuppression. The combination of cidofovir treatment with the withdrawal of immunosuppression was the most successful regimen for increasing survival rates. Survival was clearly correlated with the clearance of virus and increased titers of MAV-1-specific antibodies in sera. In addition, the passive transfer of MAV-1-specific immunoglobulin G into MAV-1-infected SCID BALB/c mice caused a marked delay in mortality, the extent of the delay being dependent on the titer of MAV-1-specific antibodies. Based on the critical role of the humoral immune response in the early defense against disseminated adenovirus infection, the concomitant use of adenovirus-specific immunoglobulins and antiviral therapy should be considered for transplant patients at risk for severe adenovirus infections.

Mouse adenovirus type 1 attachment is not mediated by the coxsackie-adenovirus receptor.

Common human adenovirus (Ad) vectors are derived from serotype 2 or 5, which use the coxsackie-adenovirus receptor (CAR) as their primary cell receptor. We investigated the receptor usage of mouse adenovirus type 1 (MAV-1), which in vivo is characterized by a pronounced endothelial cell tropism. Alignment of the fiber knob sequences of MAV-1 and those of CAR-using adenoviruses, revealed that amino acid residues, critical for interaction with CAR, are not conserved in the MAV-1 fiber knob. Attachment of MAV-1 to Chinese hamster ovary (CHO) cells was not increased by stable transfection with mouse CAR, whereas the binding efficiency of Ad2 was 20-fold higher in the mouse CAR-transfectant compared to the wild type cells. Also, purified fiber knob of Ad5, which is interchangeable with the Ad2 fiber knob, did not compete with MAV-1 for receptor binding, indicating that MAV-1 binds to a receptor different from CAR. These results support further exploration of an MAV-1-derived vector as a potential vehicle for gene delivery to cell types which are not efficiently transduced by human adenovirus vectors.

Antiviral therapy for adenovirus infections.

The treatment of severe adenovirus keratoconjunctivitis and life-threatening adenovirus infections in immunocompromised patients is still unsatisfactory. We here review the mode of action and antiviral data for cidofovir and ribavirin, obtained in cell culture, animal models or patients. Several nucleoside or nucleotide analogues have been described that target the adenovirus polymerase, whereas other antiviral targets have been poorly investigated. Furthermore, optimal therapeutic response may be achieved by combining antiviral therapy with immunotherapeutic approaches, as currently being explored.

Mouse adenovirus type 1 infection in SCID mice: an experimental model for antiviral therapy of systemic adenovirus infections.

The importance of human adenovirus infections in immunocompromised patients urges for new and adequate antiadenovirus compounds. Since human adenoviruses are species specific, animal models for systemic adenovirus infections rely on a nonhuman adenovirus. We established mouse adenovirus type 1 (MAV-1) infection of BALB/c SCID mice as a model for the evaluation of antiadenovirus therapy. In vitro studies with mouse embryonic fibroblasts pointed to the acyclic nucleoside phosphonate cidofovir and the N-7-substituted acyclic derivative 2-amino-7-(1,3-dihydroxy-2-propoxymethyl)purine (S-2242) as markedly active compounds against MAV-1. SCID mice, infected intranasally with MAV-1, developed a fatal disseminated infection after approximately 19 days, characterized by hemorrhagic enteritis. Several techniques were optimized to monitor viral, immunological, and pathological aspects of MAV-1 infection. Real-time PCR quantification of viral DNA revealed that after replication in the lungs, virus disseminated to several organs, including the brain, liver, spleen, intestine, heart, and kidneys (resulting in viruria). Immunohistochemical staining showed that MAV-1 was localized in the endothelial cells of the affected organs. Using reverse transcription-PCR, tissue levels of proinflammatory cytokines (i.e., interleukin-1beta and tumor necrosis factor alpha) were found to be markedly increased. The MAV-1/SCID model appears to be an appropriate model for in vivo evaluation of antiadenovirus agents. Treatment with cidofovir or S-2242 at a dose of 100 mg per kg of body weight resulted in a significant delay in MAV-1-related death, although these antivirals were unable to completely suppress virus replication despite continued drug treatment. These findings suggest that complete virus clearance during antiviral therapy for disseminated adenovirus infection may require an efficient adaptive immune response from the host.

Antiadenovirus activities of several classes of nucleoside and nucleotide analogues.

The absence of any formally licensed antiadenovirus drugs and the increasing incidence of life-threatening adenovirus infections in immunosuppressed patients warrant the development of effective antiadenovirus compounds. A detailed study was performed on the antiadenovirus activities of several classes of nucleoside and nucleotide analogues in human embryonic lung fibroblast cells. The antiadenovirus activities were evaluated by three methods, viz., evaluating the adenoviral cytopathic effect, monitoring cell viability by a colorimetric assay, and real-time PCR quantitation of viral DNA as a direct parameter for virus replication. The most active and selective compounds were the acyclic nucleoside phosphonate analogues cidofovir, its adenine analogue (S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine [(S)-HPMPA], and the new derivative (S)-2,4-diamino-6-[3-hydroxy-2-(phosphonomethoxy)propoxy]pyrimidine [(S)-HPMPO-DAPy]; the N7-substituted acyclic derivative 2-amino-7-(1,3-dihydroxy-2-propoxymethyl)purine (S-2242); and the 2',3'-dideoxynucleoside analogues zalcitabine and alovudine. No antiadenovirus activity was observed for the antiviral drugs ribavirin, foscarnet, acyclovir, penciclovir, and brivudin, while ganciclovir displayed modest activity. However, in human osteosarcoma cells transfected with herpes simplex virus thymidine kinase, ganciclovir demonstrated highly potent antiadenovirus activity, suggesting that the efficacy of ganciclovir against adenovirus is limited by inefficient phosphorylation in adenovirus-infected cells, rather than by insufficient inhibition at the viral DNA polymerase level. Collectively, our antiviral data show that the adenovirus DNA polymerase exhibits sensitivity to a relatively broad spectrum of inhibitors and should be studied further as an antiviral target in antiadenovirus drug development programs.

Cerebral venous thrombosis and procoagulant factors--a case study.

Cerebral venous thrombosis is a polymorphic clinical entity for which diagnosis has become more frequent with the advent of neuroradiology. The superior sagittal and transverse sinuses are frequently involved, whereas cavernous sinus thrombosis is much less frequent. Inherited resistance to the anticoagulant action of activated protein C (APC resistance), antithrombin deficiency, protein C and S deficiencies, and hyperhomocysteinemia seem to represent major causes of thrombophilia when unusual thromboembolic events (ie, before the age of 45 years) are observed. The authors present the combined occurrence of protein C and protein S deficiencies in a 32-year-old woman, manifested by extensive cerebral venous thrombosis.

Airflow obstruction and monocyte TNF release in coal workers.

Respiratory health of 102 retired coal miners was assessed by chest radiographs, lung function measurements, and questionnaires, and related to tumor necrosis factor-alpha (TNF-alpha) production by blood monocytes upon priming with different stimuli. The objective was to assess a possible relationship between airflow obstruction and TNF-alpha production in retired coal workers. No significant differences in lung function were observed between cases of coal workers' pneumoconiosis (CWP) (n = 27; > %) and references (n = 75; = > %), nor was the effect of cumulative exposure on flow volume or impedance parameters significant. TNF-alpha release upon stimulation of blood monocytes with coal mine dust was significantly increased in cases with International Labour Organisation (ILO) score 0/1 (doubtful cases) compared to references and cases with a higher ILO score. Airflow limitation defined either as a FEV1 < 80% (N = 10; 5 cases of CWP) or as a resonance frequency > 15 Hz accompanied by a negative frequency dependence of resistance (N = 9; 4 cases of CWP) was significantly related to high levels of TNF-alpha release upon stimulation with endotoxin and silica, with silica showing the strongest relation. These data suggest that in this group airflow limitation is associated with an increased expression of inflammatory mediators indifferent of the presence of pneumoconiosis.

Molecular basis for differences in susceptibility to coal workers' pneumoconiosis.

It is generally accepted that there are differences among workers in susceptibility towards the effects of mineral dusts such as silica, coal dust and asbestos. Basic research continues to find new factors involved in the process of pulmonary fibrosis caused by these minerals. In this paper, two hypotheses implicitly generated by recent findings were tested in two case-control studies among coal miners: generation of tumour necrosis factor (TNF) by blood monocytes of miners with coal workers' pneumoconiosis (CWP) and serum type III procollagen peptide (PIIIP) in CWP. Our data indicate that both parameters can be used as biological markers for early diagnosis of CWP. A follow-up study is described in which the predictive power of increased TNF release and serum PIIIP as risk factor to develop lung fibrosis will be assessed.

Acutely symptomatic benign mediastinal cysts.

Usually, cystic mediastinal masses are considered as benign. However, the size of the cyst is of importance, chiefly in a closed space such as the superior mediastinum. Rarely a dramatic symptomatology may develop though this was the case in the two patients we describe, who were admitted in the department. In the first case (a parathyroid cyst), the symptoms were due to a thrombosis of the left innominate vein, and in the second case (a thyroid cyst), the severity was dominated by a dramatic compression of the trachea and the vessels. The contribution of computed tomography is nowadays undisputed. It enables the diagnosis of the cystic nature before surgery. The diagnosis can easily be confirmed by percutaneous drainage.

[Thyroid gland and female sexual function. I. Relation outside of pregnancy]. / Glande thyroïde et fonction sexuelle féminine. I. Relations en dehors de la grossesse.

After having reviewed briefly the main characteristics of adult thyroid function, as well as the ways in which this is controlled (the first part), the authors reviewed present day methods of investigating thyroid gland function (the second part). In the third part of the work hormonal interrelationships between the thyroid and the ovary are discussed as well as the influence of gonadal hormones on thyroid function and the influence of the latter on ovarian function and on hypothalamic-pituitary interactions. The effects of thyroid diseases on reproductive function are studied in the last part of the work.