An Evaluation of Orcein Methods for Demonstrating Hepatitis B Surface Antigen and Copper-Associated Protein in Human Liver

Difficulties were encountered with the orcein method currently being used to demonstrate hepatitis B surface antigen and copper-associated protein in the liver when a new batch of dye was introduced. A survey of published material produced a plethora of methods with many contradictory recommendations. A number of methods and a variety of orceins were compared to determine which methods and orcein solutions would give the most consistent results. Two methods gave equally satisfactory results and can be recommended for routine use in screening paraffin sections of liver for hepatitis B surface antigen and copper-associated protein.     

Cyclic-AMP binding to the cell surface during development of Dictyostelium discoideum

Abstract. Cell aggregation in Dictyostelium discoideum is a chemotactic process mediated by cyclic adenosine monophosphate (CAMP), which is detected by cell surface receptors. The cAMP signal is degraded by cAMP phosphodiesterase. The possibility that cAMP signals are also used for cell communication in the multicellular stages was studied by determining whether the cAMP receptors, which are essential for signal transduction, continue to function in these stages. During slug migration, the number of binding sites per cell decreases to about 15% of the maximum level acquired during aggregation. At the onset of fruiting body formation, a three- to Four-Fold increase in cAMP binding activity occurs. This increase coincides with an increase in cAMP phosphodiesterase. Both phenomena suggest that cell-cell communication mediated by cAMP is used during culmination. During both slug migration and early culmination, the prestalk cells exhibit about twice as much binding activity as the prespore cells.     

Survival in sinoatrial disorder (sick-sinus syndrome).

A total of 381 patients with established (156) or potential (225) sinoatrial dysfunction were included in a 10-year prospective survey to determine the course of the disease and the benefits of pacing. With the exclusion of nine patients who were lost to follow-up, 61 were fitted with pacemakers. The overall survival of patients with established and potential dysfunction was similar and apparently indistinguishable from that of the normal population. Pacemaker implantation had little discernible effect on mortality though it reduced some incapacitating symptoms. These findings suggest that sinoatrial dysfunction is a relatively benign condition. Hence pacing should probably not be adopted as a routing measure but be reserved for patients with troublesome symptoms.     

Interaction of hydrophobic bis (D-mannose) derivatives with adipocyte and erythrocyte sugar transport systems

The inhibition of sugar uptake by a series of hydrophobic bis(D-mannose) derivatives has been measured in rat adipocytes. When the D-mannose moieties of the bis compounds are separated by a hexane bridge the transport inhibition constant (Ki) is greater than for a decane-bridged molecule. This is probably due to the increased hydrophobicity of the bridge of the decane-bridged compound. The enhancement in affinity due to the second sugar in the bis(D-mannose) derivatives is probably only 2-fold, since half reduction of the bis(D-mannosyloxy)hexane increases Ki approx. 2-3-fold. N'-DNP-1,3-bis(D-mannos-4'-yloxy)propyl-2-amine has very high affinity in insulin-treated cells. The affinity is approx. 1000-fold higher than for D-mannose. This enhancement is probably due to the hydrophobicity of the DNP group. The distance from the sugar to the hydrophobic group is important because an increase in Ki occurs if an aminocaproyl spacer is introduced between the DNP group and 1,3-bis(D-mannos-4'-yloxy)propyl-2-amine. Aminocaproyl and glycyl spacers also increase the Ki for NAP derivatives of 1,3-bis(D-mannos-4'-yloxy)propyl-2-amine. Each of the hydrophobic bis(D-mannose) derivatives has a lower Ki in insulin-treated cells. This may be due to an insulin responsive hydrophobic interaction between the hydrophobic portion of the sugar and a hydrophobic domain in the transport system. The inhibition constants for the hydrophobic bis(D-mannose) compounds have also been measured in human erythrocytes.     

Identification of Medically Actionable Secondary Findings in the 1000 Genomes

The American College of Medical Genetics and Genomics (ACMG) recommends that clinical sequencing laboratories return secondary findings in 56 genes associated with medically actionable conditions. Our goal was to apply a systematic, stringent approach consistent with clinical standards to estimate the prevalence of pathogenic variants associated with such conditions using a diverse sequencing reference sample. Candidate variants in the 56 ACMG genes were selected from Phase 1 of the 1000 Genomes dataset, which contains sequencing information on 1,092 unrelated individuals from across the world. These variants were filtered using the Human Gene Mutation Database (HGMD) Professional version and defined parameters, appraised through literature review, and examined by a clinical laboratory specialist and expert physician. Over 70,000 genetic variants were extracted from the 56 genes, and filtering identified 237 variants annotated as disease causing by HGMD Professional. Literature review and expert evaluation determined that 7 of these variants were pathogenic or likely pathogenic. Furthermore, 5 additional truncating variants not listed as disease causing in HGMD Professional were identified as likely pathogenic. These 12 secondary findings are associated with diseases that could inform medical follow-up, including cancer predisposition syndromes, cardiac conditions, and familial hypercholesterolemia. The majority of the identified medically actionable findings were in individuals from the European (5/379) and Americas (4/181) ancestry groups, with fewer findings in Asian (2/286) and African (1/246) ancestry groups. Our results suggest that medically relevant secondary findings can be identified in approximately 1% (12/1092) of individuals in a diverse reference sample. As clinical sequencing laboratories continue to implement the ACMG recommendations, our results highlight that at least a small number of potentially important secondary findings can be selected for return. Our results also confirm that understudied populations will not reap proportionate benefits of genomic medicine, highlighting the need for continued research efforts on genetic diseases in these populations.     

Synthesis and storage of a neurohormone in insect brains in vitro

The build-up of neurosecretory material in the median neurosecretory cells and fibre tracts of cultured cockroach brains was demonstrated by staining and bioassay. Examination of the cultured brains by electron microscopy showed active production of neurosecretory granules after 3 days in vitro. The close correlation of the results obtained by these different methods of assay leaves little doubt that a neurohormone is being synthesized and stored.