Inter-laboratory study of human in vitro toxicogenomics-based tests as alternative methods for evaluating chemical carcinogenicity: a bioinformatics perspective.

The assessment of the carcinogenic potential of chemicals with alternative, human-based in vitro systems has become a major goal of toxicogenomics. The central read-out of these assays is the transcriptome, and while many studies exist that explored the gene expression responses of such systems, reports on robustness and reproducibility, when testing them independently in different laboratories, are still uncommon. Furthermore, there is limited knowledge about variability induced by the data analysis protocols. We have conducted an inter-laboratory study for testing chemical carcinogenicity evaluating two human in vitro assays: hepatoma-derived cells and hTERT-immortalized renal proximal tubule epithelial cells, representing liver and kidney as major target organs. Cellular systems were initially challenged with thirty compounds, genome-wide gene expression was measured with microarrays, and hazard classifiers were built from this training set. Subsequently, each system was independently established in three different laboratories, and gene expression measurements were conducted using anonymized compounds. Data analysis was performed independently by two separate groups applying different protocols for the assessment of inter-laboratory reproducibility and for the prediction of carcinogenic hazard. As a result, both workflows came to very similar conclusions with respect to (1) identification of experimental outliers, (2) overall assessment of robustness and inter-laboratory reproducibility and (3) re-classification of the unknown compounds to the respective toxicity classes. In summary, the developed bioinformatics workflows deliver accurate measures for inter-laboratory comparison studies, and the study can be used as guidance for validation of future carcinogenicity assays in order to implement testing of human in vitro alternatives to animal testing.

Anti-SSA/Ro52 autoantibodies in scleroderma: results of an observational, cross-sectional study.

OBJECTIVES: To date, the diagnostic utility of anti-SSA/Ro52 autoantibodies in scleroderma and the association of them with certain clinical manifestations, particularly inflammatory myositis, are still controversial. This paper aims to assess the correlation between the presence of anti-SSA/Ro52 antibodies and the demographic, clinical and prognosis characteristics of patients with systemic sclerosis (SSc). METHODS: This is a retrospective, cross-sectional and observational study in patients with SSc. Baseline demographic and clinical characteristics were recorded. Presence of anti-SSA/Ro52, anti-SSA/Ro, anti-SSB/La, snRNP/Sm, anti-centromere, anti-Scl-70 and anti-PM-Scl were analysed by immunoblot, and antinuclear antibodies (ANA) by indirect immunofluorescence. Statistical analysis was performed with PASW Statics 18 software. RESULTS: A total of 132 consecutive patients with analysis of anti-SSA/Ro52 antibodies were selected from a Spanish cohort of 408 patients with SSc, 87.1% of them being women. About half of patients had the limited form (51.5%), followed by diffused form (18.9%), sclerosis sine scleroderma (22.7%), and pre-scleroderma (6.8%). Prevalence of anti-SSA/Ro52 was 35.6%. No association between anti-SSA/Ro52 and clinical manifestations was found, while detection of anti-SSA/Ro52 was significantly associated with the presence of anti-Ro. CONCLUSIONS: The results of our study show that anti-SSA/Ro52 antibodies are often found in SSc patients. No clinical manifestations, including inflammatory myopathy, were related with anti-SSA/Ro antibodies.

Anti-ganglioside antibodies in patients with systemic lupus erythematosus and neurological manifestations.

INTRODUCTION: Anti-ganglioside antibodies (AGA) have been associated with several peripheral neuropathies, such as Miller-Fisher syndrome, Guillain-Barré syndrome and multifocal motor neuropathy. They have also been studied in patients with systemic lupus erythematosus (SLE), focusing on neuropsychiatric manifestations and peripheral neuropathy, but the results are contradictory. OBJECTIVE: To study the presence of AGA in a large cohort of patients with SLE and neuropsychiatric manifestations. PATIENTS AND METHODS: Serum from 65 consecutive patients with SLE and neuropsychiatric manifestations, collected from 1985 to 2009, was tested for the presence of AGA antibodies (GM1, GM2, GM3, asialo-GM1 GD1a, GD1b, GD3, GT1b, GQ1b) using a standard enzyme-linked immunosorbent assay ELISA test (INCAT 1999) and thin layer chromatography (TLC). RESULTS: Positive results for asialo-GM1 (IgM) were found in 10 patients, 6 were positive for asialo-GM1 (IgM and IgG), and 4 were positive for other AGA such as GM1, GM2, GM3, GD1b, GT1b, GD3, (mainly IgM). CONCLUSIONS: Clinical and statistical studies showed no correlation between AGA and neuropsychiatric manifestations of SLE. Although some patients showed reactivity to AGA, these antibodies are not a useful marker of neuropsychiatric manifestations in SLE patients.

Echocardiography at diagnosis of antiphospholipid syndrome provides prognostic information on valvular disease evolution and identifies two subtypes of patients.

The evolution of valvular disease in antiphospholipid syndrome (APS) is barely known. In order to evaluate whether the presence or absence of valvular disease at the time of diagnosis of APS, assessed by an initial echocardiogram, predicts its subsequent evolution, we performed a prospective cohort study. We included 53 patients with APS. An initial transthoracic echocardiogram was performed on patients at the time of diagnosis of APS. Serial echocardiograms were conducted along a 12-year follow-up. Final echocardiograms were used for comparative purposes. We started with 29 patients (54%) with and 24 (45%) without valvulopathy at initial echo. At the final echocardiogram, 27 of 29 patients with initial valvulopathy continued to have valvular disease (a 93% observed likelihood), and 22 of 24 patients without initial valvulopathy demonstrated an absence of valvular disease (a 91% observed likelihood). Patients with valvulopathy in comparison with those without presented more arterial thrombotic events (69% vs. 20%, P < 0.001), atherosclerotic risk factors (62% vs. 29%, P = 0.01), livedo (48% vs. 16%, P = 0.01) and migraine (41% vs. 12%, P = 0.02). We have identified two subtypes of APS patients with and without valvulopathy by defining differential clinical features and with little crossover in valvular involvement over a long follow-up period, giving a high prognostic value to the initial echocardiographic assessment.

Cause-specific mortality after a breast cancer diagnosis: a cohort study of 10,195 women in Girona and Tarragona.

INTRODUCTION: Evidence suggests an excess of long-term mortality due to cardiovascular diseases, second tumours and other causes in patients diagnosed with invasive breast cancer (BC). Our aim was to assess this risk of death in a cohort of patients diagnosed with BC in Girona and Tarragona, northeastern Spain. MATERIALS AND METHODS: Using data from the cancer registries in these areas, a population-based cohort study was carried out including all the women diagnosed with BC during 1985-2004 and followed up until December 31st 2014 (N = 10,195). The standardised mortality ratios (SMRs) were calculated for causes other than BC in the cohort at 10 years (periods 1985-1994/1995-2004) and 20 years (period 1985-1994). The impact of competing causes of death in the long-term survival was evaluated through competing risk analysis. RESULTS: The SMRs at 10 and 20 years for all-cause mortality, except BC, were 1.21 and 1.22. The main causes of mortality showing statistically significant SMR at 10 years were other tumours (colon, lung, corpus uteri, ovary, and haematological), diabetes mellitus, diseases of the nervous system, cardiovascular diseases (after BC, the second competing cause of death among patients diagnosed > 69 years) and diseases of the kidney. Globally, the 10-year SMR was higher in the first period. After 20 years of follow-up (1985-1994 cohort), there were 48.5 excess deaths per 10,000 patient-years for causes other than BC. CONCLUSIONS: Women who did not die from BC at 10 or 20 years after the BC diagnosis had 20% higher risk of dying from other causes than women without BC. This excess risk must be clinically considered during 20 years after the BC diagnosis.

Assessing predicted age-specific breast cancer mortality rates in 27 European countries by 2020.

BACKGROUND: We assessed differences in predicted breast cancer (BC) mortality rates, across Europe, by 2020, taking into account changes in the time trends of BC mortality rates during the period 2000-2010. METHODS: BC mortality data, for 27 European Union (EU) countries, were extracted from the World Health Organization mortality database. First, we compared BC mortality data between time periods 2000-2004 and 2006-2010 through standardized mortality ratios (SMRs) and carrying out a graphical assessment of the age-specific rates. Second, making use of the base period 2006-2012, we predicted BC mortality rates by 2020. Finally, making use of the SMRs and the predicted data, we identified a clustering of countries, assessing differences in the time trends between the areas defined in this clustering. RESULTS: The clustering approach identified two clusters of countries: the first cluster were countries where BC predicted mortality rates, in 2020, might slightly increase among women aged 69 and older compared with 2010 [Greece (SMR 1.01), Croatia (SMR 1.02), Latvia (SMR 1.15), Poland (SMR 1.14), Estonia (SMR 1.16), Bulgaria (SMR 1.13), Lithuania (SMR 1.03), Romania (SMR 1.13) and Slovakia (SMR 1.06)]. The second cluster was those countries where BC mortality rates level off or decrease in all age groups (remaining countries). However, BC mortality rates between these clusters might diminish and converge to similar figures by 2020. CONCLUSIONS: For the year 2020, our predictions have shown a converging pattern of BC mortality rates between European regions. Reducing disparities, in access to screening and treatment, could have a substantial effect in countries where a non-decreasing trend in age-specific BC mortality rates has been predicted.

Long-term crude probabilities of death among breast cancer patients by age and stage: a population-based survival study in Northeastern Spain (Girona-Tarragona 1985-2004).

BACKGROUND: We provide population-based long-term survival indicators of breast cancer patients by quantifying the observed survival, and the probabilities of death due to breast cancer and to other causes by age and tumor stage at diagnosis. METHODS: We included a total of 10,195 female patients diagnosed before 85 years with invasive primary breast cancer in Girona and Tarragona during the periods 1985-1994 and 1995-2004 and followed-up until December 31st 2014. The survival indicators were estimated at 5, 10, 15 and 20 years of follow-up comparing diagnostic periods. RESULTS: Comparing diagnostic periods: I) the probability of death due to other causes did not change; II) the 20-year survival for women diagnosed ≤ 49 years increased 13% (1995-2004 = 68%; 1985-1994:55%), whereas their probability of death due to breast cancer decreased at the same pace (1995-2004 = 29%; 1985-1994 = 42%); III) at 10 years of follow-up, decreases in the probabilities of death due to breast cancer across age groups switched from 11 to 17% resulting in a risk of death reduction of 19% after adjusting by stage. During 1995-2004, the stage-specific 10-year probabilities of death due to breast cancer switched from: 3-6% in stage I, 18-20% in stage II, 34-46% in stage III and surpassed 70% in stage IV beyond 5 years after diagnosis. CONCLUSIONS: In our study, women diagnosed with breast cancer had higher long-term probability to die from breast cancer than from other causes. The improvements in treatment and the lead-time bias in detecting cancer in an early stage resulted in a reduction of 19% in the risk of death between diagnostic periods.

Predicting the cancer burden in Catalonia between 2015 and 2025: the challenge of cancer management in the elderly.

BACKGROUND: Developing effective cancer control programmes requires information on the future cancer burden in an ageing population. In our study we predicted the burden of cancer in Catalonia from 2015 to 2025. METHODS: Bayesian age-period-cohort models were used to predict the burden of cancer from 2015 to 2025 using incidence data from the Girona and Tarragona cancer registries and cancer mortality data from the Catalan mortality registry. Using the Bashir-Estève method, we divided the net change in the number of cases between 2015 and 2025 into changes due to population size (S), cancer risk (R) and age (A) distribution. RESULTS: By 2025, there will be 21,743 new cancer cases in men (40% aged > 74 years) and 17,268 in women (37% aged > 74 years). More than 40% of the new cases will be diagnosed among population aged 74 and older in prostate, colorectal, lung, bladder, pancreatic and stomach cancers in men, and in colorectal, pancreatic and bladder cancers and leukaemia in women. During 2015-2025, the number of new diagnoses will increase by 5.5% in men (A + R + S = 18.1% - 13.3% + 0.7% = 5.5%) and 11.9% in women (A + R + S = 12.4% - 1.1% + 0.6% = 11.9%). Overall cancer mortality rates will continue to decrease during 2015-2025. Lung cancer will be the most lethal cancer among men (N = 2705) and women (N = 1174). CONCLUSIONS: The increase in the number of cancer cases in Catalonia from 2015 to 2025 will mostly affect the elderly, prompting the need for increased collaboration between geriatricians and oncologists.

Technetium-99m-HMPAO brain SPECT in Behçet's disease.

UNLABELLED: Behçet's disease (BD) is an idiopathic multisystem disorder. Involvement of the central nervous system (CNS) occurs in 4%-48% of cases. The aim of this study was to evaluate 99mTc-hexamethyl propyleneamine oxime (HMPAO) SPECT findings in BD patients and eventually to detect CNS involvement by depicting cerebral blood flow disturbances. METHODS: Technetium-99m-HMPAO brain SPECT was performed on 33 consecutive BD patients. Qualitative and quantitative evaluation of the cortical uptake was done using an automatic program that generated 32 regions of interest (ROIs). An uptake index for each ROI was obtained. Reference values were obtained from a healthy control group (n = 20). Twenty-five patients also had an MRI study. RESULTS: Twelve of 32 patients (36%) presented with a clinical neurological disorder. SPECT and visual evaluation revealed that 17 patients (51.5%) had abnormalities; 9 of 25 MRI studies (36%) were abnormal. Using the quantitative approach for SPECT, 23 patients (69.7%) had abnormally low values. Six of 12 patients with neurological symptoms had a visually abnormal SPECT scan, whereas quantitative analysis showed abnormalities in 11 patients. Of the 21 patients with no neurological findings, 9 had abnormal SPECT results, and 12 had low uptake indexes. CONCLUSION: HMPAO brain SPECT shows high rates of cerebral blood flow abnormalities in BD patients presenting with neuropsychiatric symptoms, and it also is frequently abnormal in asymptomatic BD patients who have no abnormalities on MR scans. Compared with visual analysis, quantitative analysis detects an even higher rate of SPECT changes in BD patients.

Salivary and lacrimal gland dysfunction (sicca syndrome) after radioiodine therapy.

UNLABELLED: Salivary gland dysfunction has been described in patients undergoing radioiodine therapy but associated lacrimal gland dysfunction (sicca syndrome) has never been reported. We conducted a prospective cohort study with follow-up for up to 3 y in a tertiary care university center to determine the prevalence of sicca syndrome in patients after high-dose radioiodine treatment. METHODS: From January 1990 to December 1995, all patients undergoing radioiodine therapy (n = 79) with a standard dose of 925 MBq to 18.5 GBq (25-500 mCi) were interviewed using a standardized questionnaire to determine subjective ocular and oral dryness and were examined for objective lacrimal and salivary gland dysfunction. RESULTS: After radioiodine treatment, 32.9% of the patients reported subjective xerostomia and 25.3% reported subjective xerophthalmia in the first year of follow-up. Xerostomia persisted to the second year of follow-up in 20.3% of cases and was still present >3 y after the last dose of radioiodine in 15.2% of cases. Xerophthalmia persisted to the second year of follow-up in 17.7% of cases and was still present in the third year of follow-up in 13.9% of cases. Severe xerostomia occurred in 4 patients. Reduced salivary and lacrimal gland function was documented in 40 (50.6%) and 14 (17.7%) of the 79 cases, respectively, in the first year of follow-up. Objective xerostomia persisted in 13.9% of cases to the second year of follow-up and was still present in all patients >3 y after the last radioiodine application. Keratoconjunctivitis sicca persisted in 11 patients (13.9%) to the second year of follow-up but was only present in 6 patients (7.6%) >3 y after the last radioiodine application. Additionally, 28/79 patients (35.4%) who had a normal salivary gland scintigraphy previously showed reduced salivary gland function in the third year of follow-up. No significant dependence on cumulative treatment was found for objective xerostomia or xerophthalmia, but doses >11.1 GBq (300 mCi) were related to stage 3 dysfunction on salivary gland scintigraphy. CONCLUSION: Salivary and lacrimal gland dysfunction (sicca syndrome) is relatively frequent after radioiodine therapy. In most cases this is a transient side effect, but in some patients it may persist for a long period or appear late.

Systemic sclerosis developing in association with the use of interferon alpha therapy for chronic viral hepatitis.

In 1992 interferon alpha (IFNalpha) was approved by the FDA for the treatment of chronic viral hepatitis B and C. Since then IFNalpha has been implicated in the development of several autoantibodies as well as in the development or exacerbation of various autoimmune disorders. Herein, we describe a 47-year-old female who developed a limited form of systemic sclerosis (SSc) with lung involvement 6 months after the institution of IFNalpha therapy for chronic active hepatitis C. There was no family or personal history of autoimmune diseases. We speculate that the immunomodulatory effects of IFNalpha triggered the clinical manifestations of SSc in this patient. To our knowledge, this is the second case of SSc developing after therapy with IFNalpha and the first in a patient treated for chronic viral hepatitis C.

[Primary antiphospholipid syndrome: study of 27 patients]. / Síndrome antifosfolípido primario: estudio de 27 pacientes.

The clinical and biological features of a series of 27 patients with the recently described primary antiphospholipid syndrome are reported. Most of them belonged to a cohort of 90 patients who were carriers of lupus anticoagulant, which had been detected in the systematic evaluation of prolonged activated partial thromboplastin times in our hospital. Since the diagnosis they underwent a prospective protocol of follow up, with a peak follow up period of 9 years. The mean age of the 27 patients was 40.8 years and there were virtually no differences between sexes. Venous thrombosis was the most common clinical finding (16 episodes in 14 of the 27 patients). The most prevalent laboratory findings were lupus anticoagulant and IgG anticardiolipin antibodies.

[Psychoactive drug use in nursing homes]. / Utilización de fármacos para el sistema nervioso central en residencias geriátricas.

BACKGROUND: Our purpose was to know the use of psychoactive drugs including neuroleptics (NL), benzodiazepines (BNZ) and antidepressants (AD) in nursing homes (NH) in the city of Barcelona. METHOD: Cross-sectional descriptive study of 384 clinical records of people living in 19 NH. The correct use of drugs was estimated by Garrad's and Beers et al criteria. RESULTS: Average age (SD) of residents was 83 years (2.0). The average consumption of drugs was 5 (2.7); 248 individuals (64.6%) were taking at least one psychoactive drug: 81 (21%) consumed NL, 179 (46.6%) BNZ and 73 (19%) AD. 48 (12.5%) NH residents consumed long-acting BNZ and 26 (6.7%) had surpassed the recommended time of consumption for short-acting BNZ. In 21 (26%) NH residents who consumed NL, its use would not be justified. CONCLUSIONS: It is necessary to reduce the use of long-acting BNZ, to encourage a correct use of NL and to achieve a correct identification of depressive disorders in NH.

[Pulmonary thromboendarterectomy in a patient with primary antiphospholipid syndrome]. / Tromboendarterectomía pulmonar en un paciente con síndrome antifosfolípido primario.

Pulmonary arterial hypertension (PAH) is an infrequent manifestation of the primary antiphospholipid syndrome (PAPS). It may appear due to different mechanisms although the most common cause is recurrent pulmonary embolisms. In some cases the thrombi do not dissolve and organize to form fibrous masses which occlude the pulmonary veins giving place to chronic thromboembolic pulmonary hypertension. When the thrombi are located in the proximal arteries, thromboendarterectomy may be curative. The first case of a patient with PAPS diagnosed with PAH secondary to chronic thrombosis of the proximal pulmonary arteries, in whom a successful pulmonary thromboendarterectomy was performed is herein reported.

[Usefulness of cyclosporin in the treatment of pemphigus vulgaris. Apropos of 3 cases]. / Utilidad de la ciclosporina en el tratamiento del pénfigo vulgar. A propósito de 3 casos.

Cyclosporin is a useful drug in several autoimmune diseases. It has been recently evaluated, with varying results, in several dermatoses such as pemphigus vulgaris. Three cases of this condition treated with cyclosporin are reported. The drug permitted to reduce and then to suppress prolonged corticosteroid therapy, and remission was maintained after exacerbation even in monotherapy. Hypertrichosis and gum hypertrophy were the common adverse effects.

[Evaluation and significance of circulating immunocomplexes and their correlation with other immunologic parameters in connective tissue diseases]. / Estudio y significado de los inmunocomplejos circulantes y su correlación con otros parámetros inmunológicos en enfermedades del tejido conjuntivo.

The presence of circulating immunocomplexes (CIC) was evaluated in several collagen diseases and in a control group of 100 healthy individuals. Three methods were used for their detection: binding to C1q in solid phase, binding to conglutinin in solid phase, and measurement of the serum capacity to solubilize an experimental immunocomplex. In the group of patients with systemic lupus erythematosus (SLE) significant differences were found for the three techniques (p less than 0.001) and also for activity (p less than 0.001). The most sensitive method was binding to C1q. The sensitivity of the three techniques for CIC was very low in the group of patients with systemic sclerosis, and the highest rate of positive results was found with binding to C1q (10%). In the group with hypersensitivity vasculitis and polyarteritis nodosa CIC were found in 71% of cases, more than one method being positive in 50%. The highest sensitivity was obtained with the conglutinin method (48%). In patients with temporal arteritis, significant differences were only found for conglutinin binding method (p less than 0.001), with low rates of positivity.

[Multi-infarct dementia associated with antiphospholipid antibodies. Presentation of 2 cases]. / Demencia multiinfártica asociada a anticuerpos antifosfolípido. Presentación de dos casos.

Two patients with positive antiphospholipid antibody and early multi-infarction dementia as a presenting feature of their illness are reported. One was included in the so called primary antiphospholipid antibody syndrome, while the second one met the criteria for systemic lupus erythematosus. We point out to the presence of aortic regurgitation in one of the patients and its possible relation with these antibodies. Although the precise mechanism of thrombosis is incompletely known, the recognition of this type of dementia is of paramount importance as it is a potentially treatable condition.

[Cryoglobulins in systemic and rheumatological diseases. Report of 70 cases (author's transl)]. / Crioglobulinas en enfermedades sistémicas y reumatológicas. Estudio en 70 casos.

Cryoglobulins are immunoglobulins characterized by precipitating when serum is cooled and redissolving when serum is heated. There is strong evidence to consider mixed cryoglobulins as circulating immunocomplexes. Cryoglobulins have been demonstrated in association to hematologic, hepatic, lymphoproliferative, autoimmune and infectious conditions. There is also an essential or idiopathic variant. The present report studies a series of 70 patients with several rheumatic and systemic diseases, and a group of ten patients with cutaneous vasculitis. Significant levels of cryoglobulins have been detected in nine cases (overall incidence 12.8 percent). The diagnoses corresponding to these patients were as follows: systemic lupus erythematosus in three cases, dermatopolymyositis in three cases, Sjögren's syndrome in two cases, and Wegener's granulomatosis in one case. Cryoglobulins could not be demonstrated in patients with rheumatoid artritis, sclerodermia, periarteritis nodosa, cutaneous vasculitis, Reiter's syndrome, ankylosing spondilitis and acute articular rheumatism. Among patients with systemic lupus erythematosus a good correlation has been observed between the presence of serum cryoglobulins, the activity and severity of the diseases and the decrease of serum complement levels.

[Cryoglobulins and liver disease. Study of 34 cases (author's transl)]. / Crioglobulinemia en hepatopatías. Estudio en 34 pacientes.

Cryoglobulins are immunoglobulins characterized by precipitating when serum is cooled and redissolving when serum is heated. There is strong evidence to consider mixed cryoglobulins as circulating immunocomplexes, and various investigators have applied the precipitating physical property as a method to isolate immunocomplexes. In the recent years some authors have reported the presence of cryoglobulins in acute and chronic liver diseases of diverse etiology. This study investigates the presence of cryoglobulins in 34 patients with different liver diseases. Mixed cryoglobulins were detected in eight patients (23.5 percent), but only three of them had clinical symptoms attributable to the existence of cryoglobulins. In relation to the etiology of the liver disease, the highest frequency has been found among patients with hepatopathies of undetermined origin.

[Absence of antiDNA antibodies in the cryoglobulins of a case with sicca syndrome (author's transl)]. / Ausencia de anticuerpos antiDNA en las crioglobulinas de un sicca syndrome.

Cryoglobulins are serum immunoglobulins (immunocomplexes) that precipitate in the cold and redissolve on warming. Cryoglobulins from patients with several diseases showed the presence of antiDNA antibodies when previously underwent incubation in acid buffer. The possibility that antiDNA antibodies might constitute the immunocomplexes found in patients with systemic lupus erythematosus and with other connective tissue diseases is suggested. Negative findings in relation to the presence of antiDNA antibodies in cryoglobulins of a case with sicca syndrome are reported.