Utilization of fluorescence in situ hybridization with cytokeratin discriminators in TOP2A assessment of chemotherapy-treated patients with breast cancer

Tumor biomarkers increasingly provide information for predicting outcomes with chemotherapeutic regimens (personalized medicine). Topo2A is a DNA helicase targeted by anthracyclines, cytotoxic therapeutics used in both adjuvant and palliative treatments of breast cancer. TOP2A gene amplification/deletion is implicated in response to anthracycline-based chemotherapy. We describe an approach for analyzing formalin-fixed, paraffin-embedded breast tumors on tissue microarrays with TOP2A fluorescence in situ hybridization coupled with cytokeratin immunofluorescence to target tumor cells. Stained tissue from patient specimens was imaged and analyzed using Metafer/Metacyte (Metasystems, Waltham, MA, USA), including customized image classifiers. TOP2A/CEN17 ratios of 2.0 or greater (amplified) and 0.8 or less (deleted) were observed for 10.0% and 6.1% of the patients, respectively. Patient outcomes for adjuvant chemotherapy (cyclophosphamide-epirubicin-fluorouracil, cyclophosphamide-methotrexate-fluorouracil, no chemotherapy) were evaluated. No statistical significance was achieved for clinical end points regarding TOP2A status in anthracycline-treated patients. However, patients with TOP2A aberrations receiving methotrexate-based therapy exhibited a significant decrease in 5-year distant disease-free survival and breast cancer-specific overall survival, especially for patients with TOP2A deletions (disease-free survival: hazard ratio, 5.31 [P = .001], and breast cancer-specific overall survival: hazard ratio, 6.45 [P ≤ .001]). No significant differences were seen in patients included in the no-chemotherapy group. Topo2A protein levels were assessed by immunohistochemistry with no correlative statistical relevance to immunofluorescence/fluorescence in situ hybridization-based prognosis for cyclophosphamide-epirubicin-fluorouracil or cyclophosphamide-methotrexate-fluorouracil groups. Interestingly, aberrant (under)expressing patients in the no-chemotherapy group exhibited better 5-year distant disease-free survival (hazard ratio, 0.39; P = .004), trending toward more favorable breast cancer-specific overall survival (hazard ratio, 0.61; P = .11). Our results indicate a strategy by which fluorescence in situ hybridization scoring targeted to cytokeratin-positive tumor cells may provide a tool for added precision and efficiency in TOP2A evaluation from tumor tissue.     

Prevalence of faecal carriage of Carbapenemase Producing Enterobacteriaceae in healthy Indian subjects from the community

PurposeFaecal carriage of carbapenemase-producing Enterobacterales (CPE) has been extensively investigated in hospitalized patients, but limited data is available on the carriage rate in healthy individuals in India.MethodsA total of 1000 stool samples were screened for CPE from healthy individuals in Chennai (n ?= ?50), Hyderabad (n ?= ?184) and Mumbai (n ?= ?766). Diluted stool samples were cultured on chromID CARBA SMART plates. Growing colonies were screened for CPE by RAPIDEC® CARBA NP Test and minimum inhibitory concentration (MIC) of imipenem by E-Test. PCR was performed for confirmation of CPE genes.ResultsOut of the 1000 stool samples tested, 6.1% were positive for CPE. A total of 64 carbapenem resistant isolates (56 ?E.coli, 4 Klebsiella pneumoniae, 3 Enterobacter cloacae and 1 Citrobacter freundii) were recovered from ChromID CARBA SMART biplate. Carbapenemase production was identified in 57/64 isolates by RAPIDEC® CARBA NP test. PCR analysis showed 28 bla_NDM-1 and 33 bla_OXA48. Three remaining isolates (2 ?E.coli, 1 ?K.pneumoniae) were negative for the tested carbapenemase genes. Interestingly, out of these 61 PCR positive isolates, 49.1% displayed imipenem MIC within the susceptibility range on the basis of CLSI interpretative criteria.ConclusionsFaecal carriage of CPE among healthy individuals was 6.1%. Comprehensive measures to improve the sanitation scenario and implementation of National AMR action plan are needed to prevent further generation and dissemination of carbapenem resistant Enterobacterales (CRE).     

Practice patterns of physiotherapists in neonatal intensive care units: A national survey

Objective:

To determine practice pattern of physiotherapists in the neonatal intensive care units (ICUs) in India with regards to cardiopulmonary and neuromuscular physiotherapy.

Materials and Methods:

A cross-sectional survey was conducted across India, in which 285 questionnaires were sent via e-mail to physiotherapists working in neonatal intensive care units.

Results:

A total of 139 completed questionnaires were returned with a response rate of 48.7%, with a majority of responses from Karnataka, Maharashtra and Gujarat. More than 90% of physiotherapists performed chest physiotherapy in neonatal ICUs. Chest physiotherapy assessment predominantly focused on vital parameter assessment (86%) and in treatment predominantly focused on percussion (74.1%), vibration (75.5%), chest manipulation (73.3%), postural drainage (67.6%) and suction (65.4%). In neuromuscular physiotherapy more than 60% of physiotherapists used positioning, and parent education, whereas more than 45% focused on passive range of motion exercise and therapeutic handling.

Conclusion:

The practice pattern of physiotherapists for neonates in neonatal intensive care units involves both chest physiotherapy as well neuromuscular physiotherapy. Chest physiotherapy assessment focused mainly on vital parameter assessment (heart rate, respiratory rate and partial pressure of oxygen saturation SpO₂). Treatment focused on airway clearance techniques including percussion, vibration, postural drainage and airway suction. In neuromuscular physiotherapy most physiotherapists focused on parent education and passive range of motion exercise, therapeutic handling, as well as positioning.