Differential amplicons (ΔAmp)-a new molecular method to assess RNA integrity.

Integrity of the mRNA in clinical samples has major impact on the quality of measured expression levels. This is independent of the measurement technique being next generation sequencing (NGS), Quantitative real-time PCR (qPCR) or microarray profiling. If mRNA is highly degraded or damaged, measured data will be very unreliable and the whole study is likely a waste of time and money. It is therefore common strategy to test the quality of RNA in samples before conducting large and costly studies. Most methods today to assess the quality of RNA are ignorant to the nature of the RNA and, therefore, reflect the integrity of ribosomal RNA, which is the dominant species, rather than of mRNAs, microRNAs and long non-coding RNAs, which usually are the species of interest. Here, we present a novel molecular approach to assess the quality of the targeted RNA species by measuring the differential amplification (ΔAmp) of an Endogenous RNase Resistant (ERR) marker relative to a reference gene, optionally combined with the measurement of two amplicons of different lengths. The combination reveals any mRNA degradation caused by ribonucleases as well as physical, chemical or UV damage. ΔAmp has superior sensitivity to common microfluidic electrophoretic methods, senses the integrity of the actual targeted RNA species, and allows for a smoother and more cost efficient workflow.

Immersion technique for brain removal in perinatal autopsies.

Perinatal autopsies present forensic patholgists with a variety of challenges, not the least of which involves the removal and examination of very small and sometimes fragile organs. Removal of the immature brain can be particularly troublesome. Even if great care is taken during brain removal, one is often left with no more than a semifluid amorphous mass of softened tissue by the time the brain is ready to be fixed in formalin. We describe a method of perinatal brain removal which helps to preserve brain shape and integrity. By removing the brain while the head (and body) is totally immersed in water, we find that the brain is easier to remove and less apt to destruction. Subsequent fixation in formalin results in well-preserved, intact specimens, allowing for optimal examination and sectioning.

Continued heavy drinking and survival in alcoholic cirrhotics.

Survival of 64 male chronic alcoholic cirrhotics with first-onset ascites discharged from a gastrointestinal convalescent service was examined over a 32-mo period of study. Of 54 patients discharged as improved, 23 (43%) remained abstinent for a mean time of 14.0 mo and 15 (28%) resumed heavy drinking (greater than 2 g/kg/day). Twelve of 15 heavy drinkers died in a mean time of 7.2 mo, while all but one abstainer were alive. Differences in survival were statistically significantly different (p less than 0.001) when examined by the life-table method. Survival of 16 "moderate or binge" drinkers was not significantly different from abstainers. However, the number of rehospitalizations for liver disease was increased in this group. The study indicates that continued heavy drinking is associated with poor survival of alcoholic cirrhotics.

Dental infection in a cirrhotic patient. Source of recurrent sepsis.

A patient with alcoholic cirrhosis had multiple episodes of sepsis with Klebsiella pneumonia. Repeated searches for the source of infection finally revealed the organism in the root of a tooth. Evidence indicated that this site was the primary source of infection. The importance of dental infections in alcoholics and the difficulty in diagnosing those infections are emphasized by this case.