Prospective neoadjuvant analysis of PET imaging and mechanisms of resistance to Trastuzumab shows role of HIF1 and autophagy.

BACKGROUND: Although Trastuzumab has improved survival of HER2+ breast cancer patients, resistance to the agent pre-exists or develops through the course of therapy. Here we show that a specific metabolism and autophagy-related cancer cell phenotype relates to resistance of HER2+ breast cancer to Trastuzumab and chemotherapy. METHODS: Twenty-eight patients with locally advanced primary breast cancer were prospectively scheduled to received one cycle of Trastuzumab followed by a new biopsy on day 21, followed by taxol/Trastuzumab chemotherapy for four cycles before surgery. FDG PET/CT scan was used to monitor tumour response. Tissue samples were immunohistochemically analysed for metabolism and autophagy markers. RESULTS: In pre-Trastuzumab biopsies, the LC3A+/HER2+ cell population was correlated with HIF1α expression (P=0.01), while GLUT1 and LC3B expression were correlated with Ki67 proliferation index (P=0.01 and P=0.01, respectively). FDG PET tumour dimensions before therapy were correlated with LC3B expression (P=0.005). Administration of Trastuzumab significantly reduced clinical and PET-detected tumour dimensions (P<0.01). An inverse association of tumour response with the percentage of cells expressing HIF1α at baseline was documented (P=0.01). Administration of Trastuzumab resulted in a decrease of the proliferation index (P=0.004), GLUT1 (P=0.04) and HER2 (P=0.01) expression. In contrast, the percentage of LC3A+/HER2+ cells was increased (P=0.01). High baseline HIF1α expression was the only parameter associated with poorer pathological response to preoperative chemotherapy (P=0.001). CONCLUSIONS: As the HER2+/LC3A+ phenotype, which often overexpresses HIF1α, is a major subpopulation increasing after therapy with Trastuzumab, LC3A- and HIF1α-targeting therapies should be investigated for the augmentation of anti-HER2 therapy efficacy.

Mammaglobin B (SCGB2A1) is a novel tumour antigen highly differentially expressed in all major histological types of ovarian cancer: implications for ovarian cancer immunotherapy.

BACKGROUND: We studied the genetic fingerprints of ovarian cancer and validated the potential of Mammaglobin b (SCGB2A1), one of the top differentially expressed genes found in our analysis, as a novel ovarian tumour rejection antigen. METHODS: We profiled 70 ovarian carcinomas including 24 serous (OSPC), 15 clear-cell (CC), 24 endometrioid (EAC) and 7 poorly differentiated tumours, and 14 normal human ovarian surface epithelial (HOSE) control cell lines using the Human HG-U133 Plus 2.0 chip (Affymetrix). Quantitative real-time PCR and immunohistochemistry staining techniques were used to validate microarray data at RNA and protein levels for SCGB2A1. Full-length human-recombinant SCGB2A1 was used to pulse monocyte-derived dendritic cells (DCs) to stimulate autologous SCGB2A1-specific cytotoxic T-lymphocyte (CTL) responses against chemo-naive and chemo-resistant autologous ovarian tumours. RESULTS: Gene expression profiling identified SCGB2A1 as a top differentially expressed gene in all histological ovarian cancer types tested. The CD8+ CTL populations generated against SCGB2A1 were able to consistently induce lysis of autologous primary (chemo-naive) and metastatic/recurrent (chemo-resistant) target tumour cells expressing SCGB2A1, whereas autologous HLA-identical noncancerous cells were not lysed. Cytotoxicity against autologous tumour cells was significantly inhibited by anti-HLA-class I (W6/32) monoclonal antibody. Intracellular cytokine expression measured by flow cytometry showed a striking type 1 cytokine profile (i.e., high IFN-γ secretion) in SCGB2A1-specific CTLs. CONCLUSION: SCGB2A1 is a top differentially expressed gene in all major histological types of ovarian cancers and may represent a novel and attractive target for the immunotherapy of patients harbouring recurrent disease resistant to chemotherapy.

First measurement of the partial widths of 209Bi decay to the ground and to the first excited states.

209Bi alpha decay to the ground and to the first excited state have been recently observed for the first time with a large BGO scintillating bolometer. The half-life of 209Bi is determined to be τ(1/2)=(2.01±0.08)×10(19) yr while the branching ratio for the ground-state to ground-state transition is (98.8±0.3)%.

Diagnostic and prognostic impact of serum HE4 detection in endometrial carcinoma patients.

BACKGROUND: To date, no good marker for screening or disease monitoring of endometrial cancer (EC) is available. The aims of this study were to investigate HE4 gene, protein expression and serum HE4 (sHE4) levels in a panel of ECs and normal endometria (NEs) and to correlate sHE4 with patient clinicopathological characteristics and prognosis. METHODS: Using quantitative real-time PCR we tested 46 ECs and 20 NEs for HE4 gene expression. Protein expression was analysed by immunohistochemistry on tissue microarrays in 153 ECs and 33 NEs. Pre-operative serum samples from 138 EC and 76 NE patients were analysed with HE4-EIA assay. Association between sHE4 and patient clinicopathological characteristics or outcome was evaluated. RESULTS: Protein and HE4 gene were significantly upregulated in EC tissues and sera, compared with controls. High sHE4 levels were significantly associated with worse EC clinical characteristics. By univariate survival analysis, high sHE4 levels significantly correlated with decreased overall survival, progression-free survival and disease-free survival, retaining their independent prognostic value on the poorly differentiated EC cohort. CONCLUSION: We demonstrate, for the first time, that high sHE4 levels correlates with an aggressive EC phenotype and may constitute an independent prognostic factor for poorly differentiated-ECs. Determination of sHE4 could be clinically useful in identifying high-risk EC patients for a more aggressive adjuvant therapy.

Development of a combined SEM and ICP-MS approach for the qualitative and quantitative analyses of metal nano and microparticles in food products [corrected].

An integrated approach based on the use of inductively coupled plasma mass spectrometry (ICP-MS) and scanning electron microscopy (SEM) for the qualitative and quantitative analyses of metal particles in foods was devised and validated. Different raw materials and food products, like wheat, durum wheat, wheat flour, semolina, cookies, and pasta were considered. Attention was paid to the development of sample treatment protocols for each type of sample to avoid potential artifacts such as aggregation or agglomeration. The analytical protocols developed followed by ICP-MS and SEM investigations allowed us the quantitative determination and the morphological and dimensional characterization of metal nano- and microparticles isolated from the raw materials and finished food products considered. The ICP-MS method was validated in terms of linearity (0.8-80 µg/g and 0.09-9 µg/g for Fe and Ti, respectively), quantification limits (0.73 µg/g for Fe and 0.09 µg/g for Ti), repeatability (relative standard deviation (RSD) % equal to 10% for Fe and 20% in a wheat matrix as an example), and extraction recoveries (93 ± 2-101 ± 2%). Validation of the scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS) measurements was performed working in a dimensional range from 1 to 100 µm with an estimated error in the size determination equal to 0.5 µm. ICP-MS data as well as SEM measurements showed a decrease in the concentration of metal particles from wheat to flour and from durum wheat to semolina samples, thus indicating an external contamination of grains by metal particles. These findings were confirmed by environmental SEM analysis, which allowed investigation of particles of lower dimensions. Generally, the largest number of particles was found in the case of iron and titanium, whereas particles of copper and zinc were only occasionally found without any possibility of quantifying their number.

Mouse mesenchymal stem cells inhibit high endothelial cell activation and lymphocyte homing to lymph nodes by releasing TIMP-1.

Mesenchymal stem cells (MSC) represent a promising therapeutic approach in many diseases in view of their potent immunomodulatory properties, which are only partially understood. Here, we show that the endothelium is a specific and key target of MSC during immunity and inflammation. In mice, MSC inhibit activation and proliferation of endothelial cells in remote inflamed lymph nodes (LNs), affect elongation and arborization of high endothelial venules (HEVs) and inhibit T-cell homing. The proteomic analysis of the MSC secretome identified the tissue inhibitor of metalloproteinase-1 (TIMP-1) as a potential effector molecule responsible for the anti-angiogenic properties of MSC. Both in vitro and in vivo, TIMP-1 activity is responsible for the anti-angiogenic effects of MSC, and increasing TIMP-1 concentrations delivered by an Adeno Associated Virus (AAV) vector recapitulates the effects of MSC transplantation on draining LNs. Thus, this study discovers a new and highly efficient general mechanism through which MSC tune down immunity and inflammation, identifies TIMP-1 as a novel biomarker of MSC-based therapy and opens the gate to new therapeutic approaches of inflammatory diseases.

The effect of dietary soy supplementation on hot flushes.

OBJECTIVE: To assess the effect of daily dietary supplementation of soy protein isolate powder on hot flushes in postmenopausal women. METHODS: We carried out a double-blind, parallel, multicenter, randomized placebo-controlled trial of 104 postmenopausal women. Fifty-one patients (age range 48-61 years) took 60 g of products containing 40 g of isolated soy protein [corrected] daily and 53 patients (age range 45-62 years) took 60 g of placebo (casein) daily. The study lasted 12 weeks. Using analysis of covariance, we analyzed changes from baseline in mean number of moderate to severe hot flushes (including night sweats) during treatment. RESULTS: Soy was significantly superior to placebo (P < .01 in reducing the mean number of hot flushes per 24 hours after 4, 8, and 12 weeks of treatment. In particular, women taking soy had a 26% reduction in the mean number of hot flushes by week 3 and a 33% reduction by week 4 (P < .001 by the Wilcoxon exact test). By the end of the 12th week, patients taking soy had a 45% reduction in their daily hot flushes versus a 30% reduction obtained with the placebo (P < .01). The overall rates of adverse effects were similar for soy and casein-placebo. Twenty-five patients dropped out of the study: 11 in the soy group and 14 in the placebo group. Gastrointestinal side effects were the most common cause of premature withdrawal from the study (seven patients in each group). CONCLUSION: Soy protein isolate added daily to the diet substantially reduced the frequency of hot flushes in climacteric women.

Surgical correction of the septate uterus.

PIP: Metroplasty for septate uterus performed in 28 patients at Methodist Hospital in Houston is discussed. The preoperative fetal survival rate was 19.5%. Prior to laparotomy, D and C was performed in all patients. During the operation a lower abdominal transverse incision was used for abdominal exploration. The uterus was first divided from above downward in an anteroposterior direction until the endometrial cavity was entered. The incision was kept in the midline to minimize blood loss. Once the endometrial cavity was exposed, scissors were used to cut the septum bilaterally without excising it. Care was taken not to cut into the myometrium superiorly. The uterine walls were then reapproximated by simple interrupted no. 1 chromic catgut sutures that transversed serosa, myometrium, and endometrium. The first sutures were placed on the anterior and posterior aspects of the uterus just below the incision in the uterine wall. The remaining sutures were placed about .5 to 1 cm apart; none were tied until all sutures were in place. Closure was in a longitudinal anteroposterior direction along the same line as the incision. Approximation of the serosal surface was further enhanced by a running inverting 3-0 chromic catgut suture. In 16 patients, an IUD was placed in the uterine cavity before closure and left in place for 1 to 3 months. The frequency of associated endometriosis was 32.1%. Of the patients wishing to conceive, 73% became pregnant. The postoperative fetal survival rate was 87.5%. Delivery was by cesarean section.^ieng

The menopausal transition: a dynamic approach to the pathogenesis of neurovegetative complaints.

In our cross-sectional study we investigated the separate influence of three main factors, namely menopausal and estrogen status, and chronological age, on ten neurovegetative climacteric complaints reported in the scale of Kupperman et al. A multivariate statistical analysis was performed by a multivariate statistical approach on 1161 untreated women seen at the Menopause Center of the Ferrara University Hospital. Ninety women (age range, 41-54 years) were premenopausal; 492 women (age range, 38-55 years) were perimenopausal with irregular periods or amenorrhea for less than 12 months; 468 women (age range, 41-69 years) had a spontaneous menopause (age range, 37-66 years); 111 had had hysterectomy with bilateral ovariectomy while still regularly menstruating. Serum estrone was used as the indicator of the patients' estrogen status. A clear positive trend was demonstrated between menopausal status and the prevalence of depression, hot flushes, insomnia and joint pain. However, only the prevalence of hot flushes amongst these four symptoms was significantly related with the climacteric estrogen decline (beta = -0.006, P = 0.001). Moreover, menopausal status appeared to influence the intensity of fatigue, hot flushes, insomnia and paresthesia. Age was found to significantly (P = 0.053) co-vary only with the intensity of the hot flushes, with a positive relation (beta = 0.092, r = 0.104, P = 0.003), whereas estrone values did not significantly co-vary with any symptom. Furthermore, while neurovegetative symptoms are largely present also in the absence of hot flushes, when these latter are present, they exacerbate both the intensity and the prevalence of all the other symptoms.(ABSTRACT TRUNCATED AT 250 WORDS)

Sequential addition of low dose of medrogestone or medroxyprogesterone acetate to transdermal estradiol: a pilot study on their influence on the endometrium.

We evaluated bleeding pattern and endometrium following the administration of two of the most common types of progestogens used in hormone replacement therapy, medroxyprogesterone acetate (MPA) and medrogestone acetate. Twenty eight patients in spontaneous menopause were randomly allocated to two groups. Group 1 (n = 14) received 5 mg/day of of MPA and group 2 (n = 14) received 5 mg/day of medrogestone: both the progestogens were sequentially added for the last 12 days of a 21-day period of transdermal estradiol administration (50 micrograms per day). A 7-day treatment-free period completed the cycle. The study treatments were administered for 6 cycles. The endomtria were checked for their thickness by transvaginal ultrasound before starting treatment and at 6th treatment cycle (days 6-10 of the estrogen-only phase and during the period between days 8 and 12 of the progestogen addition). Endometrial biopsies were performed before starting treatment only in the patients with a positive progesterone challenge test and in all the patients at the end of the study during the addition of the progestogen. The bleeding pattern was closely monitored. MPA is accompanied by a thick endometrium with full secretory transformation in all cases. On the contrary, the same dose of medrogestone induced a consistent decrease of estrogen primed endometrium with only 4 cases of full secretory transformation. Four medrogestone-treated patients dropped out due to unscheduled bleeding. A low dose of medrogestone added to transdermal estradiol induced incomplete transformation of endometrium and oligo-amenorrhea more frequently than MPA, but it increased the chances of irregular bleeding. MPA fully transformed the endometrium: periods were thus heavier but regular. None of the patients in either group had endometrial hyperplasia.

Trazodone: a non-hormonal alternative for neurovegetative climacteric symptoms.

Hormone replacement therapy (HRT) is the treatment of choice for neurovegetative climacteric symptoms. In some women, however, HRT may either be contraindicated, or the patients themselves may prefer a non-hormonal form of treatment. Trazodone is a drug that acts a weak, but specific, inhibitor of the uptake of 3H-serotonin and is generally used for its antidepressant effects. In this study we have observed the efficacy of oral Trazodone (75 mg/day) in the treatment of the climacteric symptoms in 25 menopausal patients recruited at the Menopause Clinic of Ferrara University Hospital. The symptoms were scored from 0 to 3 according to presence and intensity. The patients were all complaining of climacteric neurovegetative symptoms (average symptom score 2.43). Symptoms scores were recorded before starting treatment and then again after 3 months. The appeared to be particularly effective on the intensity of anxiety (OR: 0.08, CI: 0-0.080), insomnia (OR: 0.15, CI: 0.02-0.71), and irritability (OR: 0.29, CI: 0.04-1.48). The intensity of hot flushes appeared reduced but was not statistically significant (OR: 0.52, CI: 0.08-1.87). However, the average total score of symptoms appearing in the Kupperman scale was reduced (-14%) after treatment. Trazodone should be kept in mind as a possible alternative to HRT. This drug can be particularly useful for those patients whose climacteric symptoms have a marked connotation of anxiety rather than for hot flushes or when HRT are contraindicated.

Effects of cyclic therapy with intranasal carbocalcitonin in healthy spontaneous postmenopausal women.

Carbocalcitonin spray administered for 12 months at a daily dosage of 80 U MRC according to five schedules has been tested on 150 normal spontaneous postmenopausal women for its influence on bone mineral density (BMD), bone metabolism and osteoarticular pain. BMD was monitored before and at the end of treatment in comparison with BMD of untreated control women. Metabolic markers (serum alkaline phosphatase, serum osteocalcin and urinary hydroxyproline) were also evaluated before and during treatment (at the 9th or 10th month of treatment). Osteoarticular pain was assessed by an analogic visual scale. Intranasal carbocalcitonin, administered according to cyclic schedules at a high frequency dosage, was able to maintain bone mass only in the earlier postmenopausal women. BMD percent increase after 12 months of treatment was 1.10 and 1.31 in women with low (< 0.870 mg/cm2) and high baseline BMD (> or = 0.870 mg/cm2), respectively. In advanced menopause the maintaining effect of carbocalcitonin on BMD seemed evident only if the baseline bone mass was lower than the BMD of the age matched control group. At least six months of treatment/year is necessary for effective therapy. Both systemic and local tolerance were optimal. No significant side-effects were detected.

[Value and current role of anterior mediastinotomy in the diagnosis of mediastinal diseases. Apropos of a series of 100 cases]. / Intérêt et place actuelle de la médiastinotomie antérieure dans le diagnostic des affections médiastinales. A propos d'une série de cent cas.

One hundred anterior mediastinotomies performed between 1981 and 1987 resulted in the diagnosis of 90 malignant mediastino-hilar or anterior mediastinal tumours, 4 benign lymph node diseases and 6 non-specific lesions. This easy to perform biopsy technique was followed by low morbidity (7%) and one death after 48 hours due to major mediastinal compression. Its sensitivity and specificity make it a valuable alternative and, according to the authors, preferable to mediastinoscopy in the diagnosis of tumours of lymph node diseases in the anterior, pulmonary hila, especially on the left side, and anterior mediastinal lymph node chains.

[Treatment of esophagotracheal fistula following intensive care. An experience of 17 cases]. / Traitement des fistules oesophago-trachéales de la réanimation (FOT). Expérience de dix-sept cas.

Oesophago-tracheal fistulae secondary to respiratory intensive care have become the commonest type of non-tumoural fistula in adults: 17/26 cases. They complicated a long period of difficult and septic intensive care, after a mean interval of 30 days. The diagnosis was confirmed by bronchoscopy which also provided good assessment of the site and extent of the fistula and the associated tracheal lesions. The severity of this complication is stressed by 5 deaths in the present series (1 toxi-infectious syndrome after 24 hours, 2 abandonments due to a hopeless resuscitation prognosis, 2 cases of oesophago-tracheal reflux which could have been prevented). One should not wait for spontaneous cure (although this did take place in 2 cases). Ideal direct suture (9 cases: 9 cures) requires local conditions favourable for healing and rapid respiratory autonomy. These conditions can be achieved by an active approach to protection against oesophago-tracheal reflux: at least lower oesophageal exclusion (4 cases) by ligation or anti-reflux operation. Oesophageal exclusion followed by oesophago-coloplasty is sometimes the only solution in cases of severe destruction of cervico-mediastinal tissues (2 cases: 1 cure). Five tracheal strictures required an associated resection-anastomosis (5 cures).

[Contribution of x-ray computed tomography and MRI to the surgical diagnosis of bronchial cancers. Comparative evaluation of x-ray computed tomography, MRI and thoracotomy in 111 cases]. / Apport respectif TDM et IRM au diagnostic d'opérabilité des cancers bronchiques. Bilan d'une confrontation TDM-IRM-thoracotomie dans 111 cas.

Computed tomography and magnetic resonance imaging have respectively decreased the incidence of exploratory thoracotomy by 11.6% to 5.7% then 5.3% after their successive inclusion in the assessment of the operability of lung cancers. Based on a systematic comparison of CT and MRI with exploratory thoracotomy in 111 patients, the yield, sensitivity and positive predictive value of these examinations was assessed in relation to tumour extension to various sites: to the chest wall, for which the sensitivity was poor (38% for CT, 54% for MRI) with a moderate predictive value (71% for CT, 77% for MRI); to the mediastinum with improved sensitivity (69% and 72%) but an uncertain predictive value (61% and 72%). The sensitivity and predictive value were then measured for lymph node involvement: N1: moderate sensitivity (69% and 76%) but a good predictive value (95% and 92%); N2 and N3: good sensitivity (79% and 93%) but a poor predictive value (70% and 66%). Apart from a few particular indications specific to MRI, especially in the apex, left hilum and in contact with the atrium, the efficacy of these two investigations is very similar. However, their lack of specificity means that certain exploratory thoracotomies are still justified in order to assess the operability of the tumour. A surgical or mediastinoscopic anatomical assessment is still necessary for a good classification of lymph node extension of lung tumours.