Dual-colour CISH is a reliable alternative to FISH for assessment of topoisomerase 2-alpha amplification in breast carcinomas

Anthracyclines are among the most powerful antineoplastic drugs available for breast cancer treatment. Although HER2 amplification has been postulated to predict anthracycline benefit, numerous reports have demonstrated that HER2/TOP2A co-amplification is the clinically useful predictive marker of response to anthracyclines. The standard technique to evaluate gene status for target therapy selection is fluorescence in situ hybridization (FISH), but this technique has some disadvantages. Dual-colour chromogenic in situ hybridization (CISH) is an extension of the FISH protocol that allows bright-field microscopy and thus represents a user-friendly alternative to FISH. In order to evaluate whether dual-colour CISH is a reliable alternative to FISH in determining TOP2A gene amplification and to determine the frequency with which TOP2A and HER2 were co-amplified, we analysed 100 invasive breast cancer specimens (70 consecutive and 30 HER2-amplified samples) using tissue microarrays. Thus, a 99 % agreement was found between TOP2A status determined by dual-colour CISH and FISH, as well as a high degree of correlation in TOP2A ratios using both techniques. TOP2A gene amplification was present in 8.6 % of the 70 consecutive samples studied, all of which were HER2-amplified. Co-amplification of TOP2A was observed in 46.5 % of the additional 30 HER2-amplified samples (no TOP2A amplification was seen in non-amplified HER2 samples). We conclude that dual-colour CISH represents an excellent alternative to FISH for determination of TOP2A gene status in invasive breast cancer. Our results showing TOP2A amplification only in HER2-amplified cases also add to the evidence that TOP2A determination should be restricted to those cases.     

Predictive factors of occult metastasis and prognosis of clinical stages I and II squamous cell carcinoma of the tongue and floor of the mouth

The incidence of occult neck metastasis in early stage tumours of the tongue and floor of the mouth varies from 20% to 30%, and the survival rates in 5 years from 73% to 97%. This study analyzes the rates of occult metastasis and prognostic factors for clinical stages I and II squamous cell carcinoma of the tongue and floor of the mouth. The records of patients with squamous cell carcinoma of the tongue and floor of the mouth, without prior treatment and treated by surgery between 1965 and 1998 were reviewed. All cases were re-staged and the surgical specimens were reviewed. This study included 193 patients, 145 men (75.1%), with ages ranging from 29 to 89 years old (mean, 60 years). The tumour site was the tongue in 132 cases (68.4%), the floor of the mouth in 45 (23.3%) and both in 16 (8.3%). With regard to stage, 85 cases were at clinical stage I (44.0%) and 108, clinical stage II (56.0%). One hundred and seventeen patients (60.6%) were submitted to a neck dissection and 27 (23.1%) had metastasic lymph nodes (pN+). The only factor associated with the presence of occult metastasis for all patients was the presence of muscular infiltration (p = 0.020); for tongue tumours the presence of vascular embolization (p = 0.043) and the presence of desmoplastic reaction (p = 0.050); for floor of the mouth tumours and T2 tumors, the histological grade (p = 0.025 and p = 0.035, respectively). Disease-free survival in 5 years was 66.4% and overall survival in 5 years 68.5%. The only factor associated with disease-free survival was the presence of muscular infiltration (p = 0.019) and with overall survival were gender (p = 0.002) and clinical stage (p = 0.031). Tumours of the tongue and floor of the mouth in the initial stages, which had muscular infiltration showed a higher probability of occult metastasis and lower disease-free survival; T2 tumours showed a worse survival as did patients of the male gender.     

Occupational risk factors for hand dermatitis among professional cleaners in Spain

Background. Dermatitis is an important health outcome for workers whose jobs put them in contact with irritants or sensitizing agents. Objectives. We conducted an analysis of data from the Epidemiological Study on the Risk of Asthma in Cleaning Workers 2 (EPIASLI2) to assess worksites and cleaning products as risk factors for hand dermatitis among professional cleaning workers. Materials/methods. We distributed 4993 questionnaires to employees of 37 cleaning companies, and used data from 818 (16%) respondents who provided information about skin symptoms and cleaning‐related exposures. We assessed associations between the frequencies of worksite and cleaning product exposures and a symptom‐based definition of hand dermatitis among current cleaning workers (n = 693) and a comparison population (n = 125). Results. Hand dermatitis was reported by 28% of current cleaning workers, versus 18% of the comparison population, and was associated with cleaning outdoor areas and schools, and the use of hydrochloric acid [prevalence ratio (PR) 1.92, 95% confidence interval (CI) 1.22–3.02] and dust mop products (PR 1.75, 95% CI 1.11–2.75). Conclusions. Professional cleaning workers may not be sufficiently protected from cutaneous disease at work. Future research should further investigate the roles of multiple product exposures and personal protective equipment.     

Endurance training modulates lymphocyte function in rats with post-MI CHF

PURPOSE: Exercise training restores innate immune system cell function in post-myocardial infarction (post-MI) rats. However, studies of the involvement of lymphocyte (Ly) in the setting of the congestive heart failure (CHF) are few. To address this issue, we investigated the function of Ly obtained from cervical lymph nodes from post-MI CHF rats submitted to treadmill running training. METHODS: Twenty-five male Wistar rats were randomly assigned to the following groups: rats submitted to ligation of the left coronary artery, which were sedentary (MI-S, N = 7, only limited activity) or trained (MI-T, N = 6, on a treadmill (0% grade at 13-20 m.m) for 60 min.d, 5 d.wk, for 8-10 wk); or sham-operated rats, which were sedentary (sham-S, N = 6) or trained (sham-T, N = 6). The incorporation of [2-C]-thymidine by Ly cultivated in the presence of concanavalin A (Con A) and lipopolysaccharide (LPS), cytokine production by Ly cultivated in the presence of phytohemagglutinin (PHA), and plasma concentration of glutamine were assessed in all groups, 48 h after the last exercise session. RESULTS: Proliferative capacity was increased, following incubation with Con-A in the MI groups, when compared with the sham counterparts. When incubated in the presence of PHA, MI-S produced more IL-4 (96%) than sham-S (P < 0.001). The training protocol induced a 2.2-fold increase in the production of interleukin-2 (P < 0.001) of the cells obtained from the cervical lymph nodes of MI-T, compared with MI-S. CONCLUSION: The moderate endurance training protocol caused an increase in IL-2 production, and a trend toward the reversion of the Th1/Th2 imbalance associated with IL-4 production increased in the post-MI CHF animal model.     

Emergence of Ciprofloxacin-Nonsusceptible Streptococcus pyogenes Isolates from Healthy Children and Pediatric Patients in Portugal

We describe 66 ciprofloxacin-nonsusceptible Streptococcus pyogenes isolates recovered from colonized and infected children. The ParC S79A substitution was frequent and associated with the emm6/sequence type 382 (emm6/ST382) lineage. The ParC D83G substitution was detected in two isolates (emm5/ST99 and emm28/ST52 lineages). One isolate (emm89/ST101) had no quinolone resistance-determining region codon substitutions or other resistance mechanisms. Five of 66 isolates were levofloxacin resistant. Although fluoroquinolones are not used in children, they may be putative disseminators of fluoroquinolone-nonsusceptible strains in the community.Streptococcus pyogenes clinical isolates with reduced susceptibility to fluoroquinolones (1, 3, 8, 14, 16, 18, 27, 29) or with high-level resistance (15, 16, 23-25, 28) have been described previously, and the reduced susceptibility to fluoroquinolones is mediated by point mutations in the quinolone resistance-determining region (QRDR) of the parC gene (3, 16, 18) whereas high-level resistance has been associated with mutations in the QRDRs of both parC and gyrA genes (23, 28). To the best of our knowledge, there are no reports documenting the prevalence and characterization of fluoroquinolone-nonsusceptible S. pyogenes associated with asymptomatic colonization. Since 1999 and 2000, we have been collecting S. pyogenes isolates from pediatric patients from different clinical origins and from carriers for the surveillance of antimicrobial susceptibility and for the epidemiological characterization of the isolates. This study aimed to describe the prevalence of ciprofloxacin-nonsusceptible S. pyogenes isolates from colonized and infected Portuguese children from 1999 to 2006 and to characterize the associated clones and resistance mechanisms.