Automated processing of fluorescence in-situ hybridization slides for HER2 testing in breast and gastro-esophageal carcinomas

Background

HER2 fluorescence in-situ hybridization (FISH) is used in breast and gastro-esophageal carcinoma for determining HER2 gene amplification and patients' eligibility for HER2 targeted therapeutics. Traditional manual processing of the FISH slides is labor intensive because of multiple steps that require hands on manipulation of the slides and specifically timed intervals between steps. This highly manual processing also introduces inter-run and inter-operator variability that may affect the quality of the FISH result. Therefore, we sought to incorporate an automated processing instrument into our FISH workflow.

Methods

Twenty-six cases including breast (20) and gastro-esophageal (6) cancer comprising 23 biopsies and three excision specimens were tested for HER2 FISH (Pathvysion, Abbott) using the Thermobrite Elite (TBE) system (Leica). Up to 12 slides can be run simultaneously. All cases were previously tested by the Pathvysion HER2 FISH assay with manual preparation. Twenty cells were counted by two observers for each case; five cases were tested on three separate runs by different operators to evaluate the precision and inter-operator variability.

Results

There was 100% concordance in the scoring between the manual and TBE methods as well as among the five cases that were tested on three runs. Only one case failed due to poor probe hybridization. In total, seven cases were positive for HER2 amplification (HER2:CEP17 ratio > 2.2) and the remaining 19 were negative (HER2:CEP17 ratio < 1.8) utilizing the 2007 ASCO/CAP scoring criteria. Due to the automated denaturation and hybridization, for each run, there was a reduction in labor of 3.5 h which could then be dedicated to other lab functions.

Conclusion

The TBE is a walk away pre- and post-hybridization system that automates FISH slide processing, improves work flow and consistency and saves approximately 3.5 h of technologist time. The instrument has a small footprint thus occupying minimal counter space. TBE processed slides performed exceptionally well in comparison to the manual technique with no disagreement in HER2 amplification status.     

Protective role of DDAH2 (rs805304) gene polymorphism in patients with myocardial infarction

The purpose of the present study was to establish the role of DDAH gene polymorphisms in the risk of developing myocardial infarction (MI) in a clinical cohort of Mexican patients. One polymorphism (rs1498373) in the DDAH1 and three in the DDAH2 (rs805304, rs3131383, and rs805305) genes were performed by TaqMan genotyping assays in 473 patients with MI and 447 healthy unrelated controls. Similar distribution of DDAH1 and DDAH2 polymorphisms was observed in MI patients and healthy controls. Under a recessive model adjusted for age, gender, and obesity, the rs805304 C allele was associated with decreased risk of MI (OR = 0.70, 95% CI = 0.51–0.96, P = 0.030). The effect of the polymorphisms on various cardiovascular risk factors was analyzed. Under a recessive model adjusted for age and gender, the DDAH2 rs805304 C allele was associated with decreased risk of obesity (OR = 0.35, 95% CI = 0.22–0.57, P = 0.001). The three DDAH2 polymorphisms were in strong linkage disequilibrium. Our results suggest that the rs805304 C allele was associated with decreased risk of MI and decreased risk of obesity.     

Fetal hydronephrosis: is there hope for consensus?

This review article aims at summarizing the data regarding fetal and neonatal hydronephrosis, at correlating controversial data with the differences in the practice of obstetrical sonography from one country to another, and finally, at presenting our own criteria for fetal renal collecting system dilatation along with our own guidelines of postnatal investigation.     

Hypertension 2008—Awareness,Understanding, and Treatment of Previously Diagnosed Hypertension in Baby Boomers and Seniors: A Survey Conducted by Harris Interactive on Behalf of the Preventive Cardiovascular Nurses Association

J Clin Hypertens (Greenwich). A Harris Interactive survey of 1548 hypertensive persons aged 44 and older confirms the findings of previous studies that showed suboptimal rates of adherence to medication and lifestyle regimens to lower blood pressure, despite a high level of awareness of the health consequences of uncontrolled blood pressure. When the study population was analyzed by age group (baby boomers, ages 44 to 62 years, and seniors, ages ≥63 years), nonadherence was greater in the baby boomer cohort, which nevertheless had a higher level of concern than the seniors. Poor communication between patients and health care providers contributes to nonadherence to treatment regimens. Patients’ age plays an important role in their attitudes and behaviors regarding illness and treatment as well as their preferences as to the types of educational materials they would find helpful and the ways those materials can best be delivered. Because of the growing population of baby boomers further studies are warranted to evaluate attitudes, knowledge, and behaviors concerning the identification and treatment of hypertension.     

Welfare reform and the health of young children: a sentinel survey in 6 US cities

CONTEXT: Welfare reform under the 1996 Personal Responsibility and Work Opportunity Reconciliation Act replaced entitlement to cash assistance for low-income families with Temporary Assistance to Needy Families, thereby terminating or decreasing cash support for many participants. Proponents anticipated that continued receipt of food stamps would offset the effects of cash benefit losses, although access to food stamps was also restricted. OBJECTIVE: To examine associations of loss or reduction of welfare with food security and health outcomes among children aged 36 months or younger at 6 urban hospitals and clinics. DESIGN AND SETTING: A multisite retrospective cohort study with cross-sectional surveys at urban medical centers in 5 states and Washington, DC, from August 1998 through December 2000. PARTICIPANTS: The caregivers of 2718 children aged 36 months or younger whose households received welfare or had lost welfare through sanctions were interviewed at hospital clinics and emergency departments. MAIN OUTCOME MEASURES: Household food security status, history of hospitalization, and, for a subsample interviewed in emergency departments, whether the child was admitted to the hospital the day of the visit. RESULTS: After controlling for potential confounding factors, children in families whose welfare was terminated or reduced by sanctions had greater odds of being food insecure (adjusted odds ratio [AOR], 1.5; 95% confidence interval [CI], 1.1-1.9), of having been hospitalized since birth (AOR, 1.3; 95% CI, 1.0-1.7) and, for the emergency department subsample, of being admitted the day of an emergency department visit (AOR, 1.9; 95% CI, 1.2-3.0) compared with those without decreased benefits. Children in families whose welfare benefits were decreased administratively because of changes in income or expenses had greater odds of being food insecure (AOR, 1.5; 95% CI, 1.1-2.2) and of being admitted the day of an emergency department visit (AOR, 2.8; 95% CI, 1.4-5.6). Receiving food stamps does not mitigate the effects of the loss or reduction of welfare benefits on food security or hospitalizations. CONCLUSION: Terminating or reducing welfare benefits by sanctions, or decreasing benefits because of changes in income or expenses, is associated with greater odds that young children will experience food insecurity and hospitalizations.     

A comparison of oral diazepam versus midazolam,administered with intravenous meperidine,as premedication to sedation for pediatric endoscopy

OBJECTIVES: This study was performed to compare the effects of oral midazolam and oral diazepam, administered with intravenous (IV) meperidine, on pre-procedural, procedural, and post-procedural sedation and recovery in children undergoing diagnostic upper endoscopy. The costs of pre-procedure sedation were compared for the two benzodiazepines. METHODS: A randomized, double-blind study was conducted in 154 children (mean age 96.73 +/- 59.34 months, 53% male) undergoing endoscopy. Oral midazolam (0.5 mg/kg, maximum dose of 20 mg) or oral diazepam (0.3 mg/kg, maximum dose of 10 mg) was given before IV insertion, and with IV meperidine (2 mg/kg, maximum dose of 100 mg) given to all patients just before upper endoscopy. Further "rescue" midazolam doses (to a maximum cumulative dose of 5 mg) were given as needed to achieve a pre-procedure sedation score of > or =2. All patients received intravenous propofol for procedural sedation. Patients were evaluated for the efficacy and safety of pre-procedural sedation, sedation during upper endoscopy, and recovery following completion of the procedure. RESULTS: There were no significant differences between study groups for level of pre-procedural sedation, need for midazolam rescue in endoscopy, effectiveness of procedural sedation, occurrence of adverse events, and recovery parameters. CONCLUSIONS: Oral midazolam and diazepam, in conjunction with IV administration of meperidine, provide comparable, effective, and safe premedication for children undergoing upper endoscopy. The cost of midazolam was substantially higher than diazepam.     

Alendronate treatment for infants with osteogenesis imperfecta: demonstration of efficacy in a mouse model

Recent non-placebo-controlled studies of the bisphosphonate pamidronate have shown it to be effective in reducing fractures and improving bone density in infants and children with osteogenesis imperfecta (OI). To evaluate the effects of bisphosphonate treatment in a controlled study, the oim/oim mouse model of OI was studied. Nursing infant mouse pups (approximately 2 wk old) with moderate to severe OI (oim/oim mouse) and age- and background-matched control mice (+/+) were treated either with the third-generation bisphosphonate alendronate (ALN), or with saline. Fracture risk, bone quality, and growth were evaluated over a 12-wk treatment period. ALN at a dose of 0.03 mg/kg/d or saline was administered via s.c. injection to infant oim/oim and wild-type (+/+) mice from 2 to 14 wk of age (n = 20 per subgroup). The average number of fractures sustained by the ALN-treated oim/oim mice was reduced significantly compared with the untreated oim/oim mice (0.7 +/- 0.7 fractures/mouse versus 2.0 +/- 0.2 fractures/mouse). Bone density increased significantly in the femur and the spine with treatment (2.0 +/- 0.5 versus 1.2 +/- 0.5 in femur and 2.1 +/- 0.5 versus1.6 +/- 0.5 in spine). Histologic evaluation revealed the percentage of metaphyseal tibial bone increased significantly with treatment in both +/+ and oim/oim mice. Mechanical testing revealed an increase in structural stiffness for both treated +/+ and oim/oim mice compared with untreated animals. None of the material properties examined were significantly altered with treatment, nor was spinal curvature affected. Weight gain and long bone growth were comparable in the treated and untreated oim/oim mice. In wild-type mice, femur lengths were significantly shorter in the treated mice compared with untreated counterparts. This animal study demonstrates that treatment of OI in mice as early as 2 wk of age with ALN appears to be effective in reducing fractures and increasing bone properties. Based on the data from this study, ALN therapy in infants with OI should prove to be effective.