Integrative Analysis of DCE-MRI and Gene Expression Profiles in Construction of a Gene Classifier for Assessment of Hypoxia-Related Risk of Chemoradiotherapy Failure in Cervical Cancer.

PURPOSE: A 31-gene expression signature reflected in dynamic contrast enhanced (DCE)-MR images and correlated with hypoxia-related aggressiveness in cervical cancer was identified in previous work. We here aimed to construct a dichotomous classifier with key signature genes and a predefined classification threshold that separated cervical cancer patients into a more and less hypoxic group with different outcome to chemoradiotherapy. EXPERIMENTAL DESIGN: A training cohort of 42 patients and two independent cohorts of 108 and 131 patients were included. Gene expression data were generated from tumor biopsies by two Illumina array generations (WG-6, HT-12). Technical transfer of the classifier to a reverse transcription quantitative PCR (RT-qPCR) platform was performed for 74 patients. The amplitude ABrix in the Brix pharmacokinetic model was extracted from DCE-MR images of 64 patients and used as an indicator of hypoxia. RESULTS: Classifier candidates were constructed by integrative analysis of ABrix and gene expression profiles in the training cohort and evaluated by a leave-one-out cross-validation approach. On the basis of their ability to separate patients correctly according to hypoxia status, a 6-gene classifier was identified. The classifier separated the patients into two groups with different progression-free survival probability. The robustness of the classifier was demonstrated by successful validation of hypoxia association and prognostic value across cohorts, array generations, and assay platforms. The prognostic value was independent of existing clinical markers, regardless of clinical endpoints. CONCLUSIONS: A robust DCE-MRI-associated gene classifier has been constructed that may be used to achieve an early indication of patients' risk of hypoxia-related chemoradiotherapy failure. Clin Cancer Res; 22(16); 4067-76. ©2016 AACR.

Interplay between promoter methylation and chromosomal loss in gene silencing at 3p11-p14 in cervical cancer.

Loss of 3p11-p14 is a frequent event in epithelial cancer and a candidate prognostic biomarker in cervical cancer. In addition to loss, promoter methylation can participate in gene silencing and promote tumor aggressiveness. We have performed a complete mapping of promoter methylation at 3p11-p14 in two independent cohorts of cervical cancer patients (n = 149, n = 121), using Illumina 450K methylation arrays. The aim was to investigate whether hyperm-ethylation was frequent and could contribute to gene silencing and disease aggressiveness either alone or combined with loss. By comparing the methylation level of individual CpG sites with corresponding data of normal cervical tissue, 26 out of 41 genes were found to be hypermethylated in both cohorts. The frequency of patients with hypermethylation of these genes was found to be higher at tumor stages of 3 and 4 than in stage 1 tumors. Seventeen of the 26 genes were transcriptionally downregulated in cancer compared to normal tissue, whereof 6 genes showed a significant correlation between methylation and expression. Integrated analysis of methylation, gene dosage, and expression of the 26 hypermethylated genes identified 3 regulation patterns encompassing 8 hypermethylated genes; a methylation driven pattern (C3orf14, GPR27, ZNF717), a gene dosage driven pattern (THOC7, PSMD6), and a combined methylation and gene dosage driven pattern (FHIT, ADAMTS9, LRIG1). In survival analysis, patients with both hypermethylation and loss of LRIG1 had a worse outcome compared to those harboring only hypermethylation or none of the events. C3orf14 emerged as a novel methylation regulated suppressor gene, for which knockdown was found to promote invasive growth in human papilloma virus (HPV)-transformed keratinocytes. In conclusion, hypermethylation at 3p11-p14 is common in cervical cancer and may exert a selection pressure during carcinogenesis alone or combined with loss. Information on both events could lead to improved prognostic markers.

High salivary secretory IgA antibody levels are associated with less late-onset wheezing in IgE-sensitized infants.

Low levels of secretory IgA (SIgA) and transient IgA deficiency have been associated with an increased risk for allergy, but data are conflicting. The aim was to assess the relationship between salivary SIgA antibody levels at 1 yr and wheezing at age four in a birth cohort, in particular the possible protective role of salivary SIgA in sensitized children. Saliva samples were obtained from all children (n=67) with a positive skin prick test (SPT) at 1 yr and 212 children with a negative SPT. In all, 200 of these children responded to questionnaires at 4 yrs and 183 were skin prick tested at that age. The levels of salivary SIgA and salivary IgA antibodies to the most common food allergen egg and inhalant allergen cat were analyzed by ELISA. Serum was analyzed for IgE antibodies to egg and cat. Development of late-onset wheezing was associated with low SIgA levels in children with positive SPT to at least one allergen both at 1 and 4 yrs of age (p=0.04), as well as in children with circulating IgE antibodies to egg or cat at 1 yr (p=0.02). None of nine persistently sensitized children with SIgA levels in the upper quartile developed wheezing, when compared to 10/20 children with lower levels (p=0.01). Older siblings, more than three infections during infancy, at least one smoking parent, and male gender, were all associated with SIgA in the upper quartile. In conclusion, high levels of SIgA antibodies in sensitized infants were associated with significantly less late-onset wheezing, supporting a protective role against development of asthmatic symptoms. Recurrent infections and other factors supporting an increased microbial pressure during infancy were associated with high levels of salivary SIgA.

Leptin and asthma in overweight children at 12 years of age.

Obesity is suggested as a risk factor for asthma, but the mechanisms are unclear. The relationship between obesity and asthma has not been considered in children born with very low-birth weight (VLBW). We hypothesized that overweight was a contributing factor for asthma in VLBW children, and that leptin and leptin-associated cytokines might play roles in overweight-related asthma. Seventy-four VLBW and 64 normal birth weight (NBW) children participated in a 12-yr follow up study assessing asthma and allergy. Twenty-seven (12 VLBW) of the 138 children were overweight according to the proposed international definition. The diagnosis of current asthma was made by a pediatrician. Serum levels of leptin and interferon (IFN)-gamma were analyzed by enzyme-linked immunosorbent assay (ELISA). Leptin levels were considerably higher in the overweight than in the non-overweight children (median value: 18.1 vs. 2.8 ng/ml, p < 0.001). In the overweight children, current asthmatics had twice as high levels of leptin as children without current asthma (median value: 30.8 vs. 14.3 ng/ml, p = 0.14), but this was not the case in the non-overweight children. IFN-gamma was more often detected in the overweight than in the non-overweight children (61% vs. 12%, p < 0.001), and there was a positive correlation between the levels of leptin and the levels of IFN-gamma (Rho = 0.40, p < 0.001). In the VLBW group, the overweight children had a significantly increased risk for current asthma compared with the non-overweight children after adjustment for the neonatal risk factors [adjusted odds ratio (OR) 5.8, 95% confidence interval (CI): 1.2-27]. Thus, overweight was associated with asthma in the VLBW children. Our hypothesis remained that leptin might be involved in the pathogenesis of asthma in the overweight children, and IFN-gamma might be a pathway in the process of leptin-induced inflammation.

Mammary epithelial paracellular permeability in atopic and non-atopic mothers versus childhood atopy.

Sodium/potassium (Na/K) ratios are considered to be a marker of mammary epithelial paracellular permeability. The aim of the present study was to investigate the association between maternal atopy and Na/K ratios in breast milk and the association between Na/K ratios in breast milk and the development of atopy in the offspring. Early and mature milk samples were obtained from 30 atopic and 43 non-atopic women. We found no differences in the Na/K ratios between atopic and non-atopic women. At 18 months of age, 22 (30%) of the children had a positive skin prick test (SPT) and 26 (36%) had symptoms of atopic diseases. Overall, high levels of Na/K compared with low and slightly raised levels of Na/K in the maternal milk tended to be associated with a positive SPT and atopic disease. However, if the mother was atopic, high levels of Na/K in early or mature milk were associated with a significantly increased risk of a positive SPT or atopic disease in the offspring [RR = 4.8 (1.9-12)] whereas no such association was observed in non-atopic mothers [RR = 0.8 (0.4-1.7), p for interaction = 0.001]. Thus, high Na/K levels in the breast milk may be associated with the development of atopy and atopic diseases in the offspring of atopic mothers.

Allergen-induced cytokine secretion in atopic and non-atopic asthmatic children.

Atopic asthma is characterized by excessive T helper 2 (Th2)-like immunity to allergens in the bronchial mucosa. The Th2-cytokine interleukin (IL)-4 induces IgE production, while the Th2-cytokine IL-5 promotes eosinophilic inflammation in the airways of asthmatics. Most asthmatics are atopic, but a subgroup is non-atopic. We hypothesize that allergen-induced Th2, particularly IL-5, responses can be observed in peripheral blood in both atopic and non-atopic asthmatic children but not in healthy control children. The aim of the present study was to determine IL-4, IL-5, IL-9, IL-10, IL-13 and IFN-gamma secretion induced from peripheral blood mononuclear cells (PBMC) by a broad panel of inhalant allergens (timothy, cat, birch, dog and house dust mite) in asthmatic children with and without sensitization. The study included 13 atopic asthmatic, 5 non-atopic asthmatic, and 12 non-atopic non-asthmatic children. PBMC were stimulated with allergens and cytokine production was measured with enzyme-linked immunosorbent assay (ELISA). Higher levels of cat and dog antigen-induced IL-5 release were more commonly observed in both atopic and non-atopic asthmatics than in controls. Children with atopic, but not non-atopic, asthma produced higher levels of allergen-induced IL-4 and IL-9 than controls. Non-atopic asthmatics produced more IL-10 than atopic asthmatics after cat stimulation. High levels of eosinophilia-associated IL-5 responses are induced by cat and dog allergen in both atopic and non-atopic asthmatic children. The Th2 cytokines IL-4 and IL-9 were associated only with atopic asthma, probably due to their IgE-inducing properties.

Effects of breast milk from allergic and non-allergic mothers on mitogen- and allergen-induced cytokine production.

Breast milk contains several components that provide specific immunity and affect the maturation of the infant's immune system. The aim of this study was to analyze the effects of breast milk, on mitogen- and allergen-induced cytokine production from cord blood mononuclear cells (CBMC), and if those effects differ between allergic and non-allergic mothers. The cells were incubated for 96 h with phytohemagglutinin (PHA), ovalbumin or cat dander in the presence of various dilutions of colostrum. Colostrum inhibited both mitogen- and cat-induced IFN-gamma and mitogen-induced interleukin-4 (IL-4) production. The inhibition on IFN-gamma production was to some extent caused by TGF-beta, as the effect was modified when an anti-TGF-beta antibody was added to the cultures. In contrast, colostrum enhanced allergen-induced production of the Th2-like cytokines IL-5 and IL-13, and this was accompanied with increased production of IL-10. No differences were found between allergic and non-allergic mothers. The inhibitory effect of breast milk on IFN-gamma production, which was partly due to the high levels of TGF-beta, together with the enhancing effect on IL-10 secretion, confirm that breast milk is anti-inflammatory. Although the production of IL-5 and IL-13 was enhanced by colostrum, this was accompanied with an increased production of IL-10. Together with the high levels of TGF-beta in breast milk and inhibitory effect of colostrum on IL-4 production, this suggests a possible mechanism whereby breast-feeding may protect against the development of allergy. Despite differences in the composition of breast milk between allergic and non-allergic mothers, the effects of breast milk on cytokine production from CBMC were independent of the atopic status of the mothers.

Cytokine, chemokine and secretory IgA levels in human milk in relation to atopic disease and IgA production in infants.

The relationship between breast-feeding, IgA production and development of atopic disease in children is a matter of controversy. Some of this controversy might be due to individual differences in the composition of breast milk. The aim of this study was to relate the levels of cytokines, chemokines and secretory (S)-IgA antibodies in breast milk to the development of atopic manifestation and salivary IgA production in infants. Cytokine, chemokine and SIgA levels, as measured with enzyme-linked immunosorbent assay (ELISA), in colostrum and mature milk were analyzed in relation to the development of positive skin-prick tests (SPT), allergic symptoms and salivary IgA antibody production during the first 2 years of life in 53 infants. There was no association between levels of IL-4, -5, -6, -8, -10, -13, -16, IFN-gamma, TGF-beta1, -beta2, RANTES, eotaxin or SIgA levels in the breast milk with either SPT-positivity, development of allergic symptoms or salivary IgA levels during the first 2 years of life in the infants. Thus, differences in the composition of cytokines, chemokines and SIgA in breast milk did not, to any major degree, affect the development of a positive SPT, atopic symptoms, nor salivary IgA antibody production during the first 2 years of life.

Enhancement of hypoxemia by right-to-left atrial shunting in severe asthma.

We report two cases of severe hypoxemia due to right-to-left shunt in acute asthmatic patients. During acute asthma, the transmural right atrial pressure can be higher than left atrial pressure during inspiration and then induce a right-to-left shunt through inter-atrial communication leading to hypoxemia. Contrast echography as well as Doppler analysis can easily confirm the diagnosis.

[Rhabdomyolysis in community-acquired pneumonia]. / Rhabdomyolyses au cours des pneumopathies communautaires.

Several cases of rhabdomyolysis associated with pneumococcal pneumonia have been recently reported. However their significance have been poorly investigated. In this retrospective study, we have compared the patients admitted in ICU for acute community-acquired pneumonia with or without rhabdomyolysis (group A: CPK > or = 1,000 Ul/l and group B: CPK < 1,000 Ul/l). Among the 41 patients of the study, 12 (29%) belonged to the group A (CPK = 1,852 +/- 535 Ul/l) and 29 (71%) to group B (CPK = 190 +/- 35 Ul/l). Neither the initial severity, nor renal impairment were different in the two groups. However the mortality was significantly higher in the group A (4/12) than in group B (2/29) (P < 0.05). In this study, pneumonia is frequently associated with rhabdomyolysis, without etiologic significance. In patients with pneumonia, rhabdomyolysis seems to be a bad prognostic indice unrelated with renal impairment.

[A fatal case of viper bite in an adult in France]. / Un cas mortel de morsure de vipère chez un adulte en France.

Vipera bites in France are supposed to be benign. We report here, the case of a snake bite with fatal outcome due to hypoxemia and shock, occurring in a young woman without previous disease. This case could lead to reevaluate the indication of antivenomous serotherapy in the severe envenomations. However the tolerance of the new serums has to the confirmed.

Severe non-hypoxic bradycardia during disconnection from the ventilator during the recovery phase of ARDS.

Four patients with adult respiratory distress syndrome developed sinus bradycardia during weaning procedures with no evidence of hypoxemia. Bradycardia occurred immediately after the patients' endotracheal tubes were disconnected from the ventilator and most of the time resolved after reconnection. However, 3 patients eventually deteriorated, requiring advanced life support and for one of them, cardiac pacing. The precise mechanism of these bradycardic episodes remains unclear but was unrelated to hypoxemia or acid-base disorder.

[Continuous hemofiltration in the treatment of acute kidney failure in multiple organ failure]. / Hémofiltration continue dans le traitement de l'insuffisance rénale aiguë au cours de défaillance polyviscérale.

Twenty-nine patients with acute renal failure and multiple organ failure were treated with continuous arterio-venous or veno-venous haemofiltration for a period of 6 +/- 5 days. Sixteen of these patients were improved and haemofiltration was withdrawn, but secondary worsening occurred in 8 cases. The method is well tolerated by the cardiovascular system and provides a satisfactory metabolic control. The low patient's survival rate (27 percent) is due to the severity of the disease. In the absence of controlled studies, it is difficult to assert that continuous haemofiltration is better than conventional haemodialysis.

[Spontaneous course of myxoma of the left atrium]. / Evolution spontanée du myxome de l'oreillette gauche.

Little is known about the natural history of left intra-atrial myxomas. We report 3 cases of that disease where successive echocardiographic examinations provided figures of 11, 12 and 14 months respectively for the formation of the tumour in the left atrium. Echocardiography is perfectly reliable for the diagnosis of myxoma. False-negative results are rare and usually due to very small myxomas being beyond the resolution potential of the instrument; this seems to have been the case in 2 of our 3 patients. Two data, however, remain unknown: the beginning of formation and the rapidity of tumoral growth. These 3 cases raise the problem of repeat echocardiography some time after a cerebral accident of suspected embolic origin, when the initial examination is negative.

Sequential myocardial scintigraphy with technetium-99m stannous pyrophosphate following myocardial infarction.

Studies have shown that technetium-99m stannous pyrophosphate (Tc-PPi) is effective for the detection and imaging of acute myocardial infarction. Positive Tc-PPi myocardial scintigrams, however, have been reported in patients with other forms of heart disease and no evidence of recent myocardial infarction. To help define the usefulness of this test, we undertook a prospective study to ascertain when Tc-PPi myocardial scintigrams return to normal after myocardial infarction. Twenty patients with acute myocardial infarction were followed with Tc-PPi scintigrams at 1 and 2 wk, and 1, after infarction. The serial scintigrams revealed that a) 15 of 18 scintigrams were positive within the first week after infarction, b) the number of markedly positive scintigrams decreased promptly after the first week, and c) some scintigrams (11 of 18 at 1 mo, and 3 of 18 at 9 mo) remained positive throughout the study. The possible explanations for persistently positive scintigrams are discussed. Persistently positive scintigrams may hinder the usefulness of Tc-PPi myocardial scintigraphy for the diagnosis of acute myocardial infarction in patients who have had a myocardial infarction within the previous 9 mo.