Microdissection genotyping of archival fixative treated tissue for Gaucher disease

The genetic diagnosis of Gaucher disease by molecular methods is complicated by the existence of a highly homologous transcribed pseudogene (96% identity) that is found in close proximity to the true gene on chromosome 1q21. In addition, the pseudogene sequence can mimic disease-causing mutations in the true gene. Selective polymerase chain reaction (PCR) amplification of the true gene can be accomplished in extracted DNA from fresh-frozen samples by designing oligonucleotide primers to hybridize to defined regions that are not present in the pseudogene. This standard molecular approach, which entails amplification of relatively long segments of intact DNA, is not feasible in archival, paraffin-embedded, solid-tissue specimens in which the negative effects of chemical fixation result in DNA strand scission and breakdown of nucleic acid. A novel approach, specifically created for use with archival, fixative-treated tissue specimens, was developed for detection and characterization of common mutations of Gaucher disease. Three separate robust PCR reactions were formulated, 2 for selective amplification of portions of only the true gene exons 2 and 9, with a third reaction targeting exon 10, wherein both the true and pseudogene were coamplified. In the latter, DNA sequencing was used to determine the presence of true and pseudogene allele content in addition to identification of base sequence alterations. This method, requiring a single, 4-microm-thick histologic section, was successfully applied to archival paraffin block tissue specimens that had been in storage for up to 75 years. It was capable of accurately genotyping common Gaucher disease mutations as well as discovering a novel mutation and genetic polymorphism. We recommend our approach when only fixative-treated tis sue is available for molecular genotyping.     

Comparison of EGS4 and MCNP4b Monte Carlo codes for generation of photon phase space distributions for a Varian 2100C

Monte Carlo based dose calculation algorithms require input data or distributions describing the phase space of the photons and secondary electrons prior to the patient-dependent part of the beam-line geometry. The accuracy of the treatment plan itself is dependent upon the accuracy of this distribution. The purpose of this work is to compare phase space distributions (PSDs) generated with the MCNP4b and EGS4 Monte Carlo codes for the 6 and 18 MV photon modes of the Varian 2100C and determine if differences relevant to Monte Carlo based patient dose calculations exist. Calculations are performed with the same energy transport cut-off values. At 6 MV, target bremsstrahlung production for MCNP4b is approximately 10% less than for EGS4, while at 18 MV the difference is about 5%. These differences are due to the different bremsstrahlung cross sections used in the codes. Although the absolute bremsstrahlung production differs between MCNP4b and EGS4, normalized PSDs agree at the end of the patient-independent geometry (prior to the jaws), resulting in similar dose distributions in a homogeneous phantom. EGS4 and MCNP4b are equally suitable for the generation of PSDs for Monte Carlo based dose computations.     

Recurrent diffuse lipomatosis of the neck--a case report

Diffuse lipomatosis is a rare disorder of adipose tissue occurring in young people . It has a predilection for trunk and proximal extremities where it presents as poorly circumscribed overgrowth of fatty tissue. Definite diagnosis is established by histological examination of tumor. Though it attains extensive size and has a high tendency to recur, the clinical course is benign.     

Patterns of coverage of psychiatric emergencies

A massive change in the detection of psychiatric cases in the emergency room was recorded when pattern of coverage was changed from "on-call" basis to "continuous physical presence" of psychiatry residents in the emergency room.     

Alterations in Breast Cancer Biomarkers Following Neoadjuvant Therapy

Annals of Surgical Oncology - Biomarker changes in patients with residual disease (RD) after neoadjuvant systemic therapy (NAT) have unclear consequences. This study examined the prevalence of...