New Developments in the Understanding and Treatment of Autoimmune Hemolytic Anemia: Traditional and Novel Tests.

Careful consideration of the clinical history with traditional testing such as an antibody screen and direct antiglobulin test (DAT) allow for the categorization of most forms of autoimmune hemolytic anemia. Based on the initial findings, specialized testing can further categorize disease entities and increase the sensitivity of testing. In this section, we explain the diagnostic findings of both traditional and novel testing and how their appropriate interpretations help distinguish the forms of autoimmune hemolytic anemia (AIHA).

ABO maternal-child discordance: Evidence of variable allelic expression and considerations for investigation.

BACKGROUND: We report a case of apparent mother-child ABO group noninheritance. A Caucasian mother initially typed as group O and her infant group AB. Investigation ruled out preanalytical causes such as mislabeled samples and in vitro fertilization. MATERIALS AND METHODS: Red blood cells were characterized by routine serologic testing. Genomic data were analyzed by targeted polymerase chain reaction-restriction fragment length polymorphism and Sanger sequencing. Transferase structures were modeled using PyMOL molecular visualization software. RESULTS: Serologic testing initially demonstrated the mother was group O, father group AB, and infant group AB. Further testing of the maternal sample with anti-A,B demonstrated weak A expression. Molecular testing revealed the maternal sample had an ABO*O.01.01 allele in trans to an A allele, ABO*AW.29 (c.311T>A, p.Ile104Asn), determined by gene sequencing. The sample from the infant carried the same ABO*AW.29 allele in trans to a B allele, ABO*B.01. CONCLUSION: ABO genotyping revealed an A transferase encoded by ABO*AW.29, with apparent variable activity. Although A antigen expression is well known to be weak in newborns, it was robust on the red blood cells (RBCs) of the AB infant and undetectable with anti-A on the mother. Variable expression of weak subgroups may reflect competition or enhancement by a codominant allele, as well as glycan chain maturation on red cells. Previous examples in group AB mothers with Aweak infants suggested that the decreased expression is primarily due to glycan immaturity. To our knowledge, this is the first reported case of the ABO*AW.29 allele presenting with weak A expression in a group Aweak mother and robust A expression in a group AB infant, suggesting the in trans allele is an important factor in determining transferase activity and may override age-related effects.

Molecular phylogeny of the genus Lasiopogon (Diptera: Asilidae) and a taxonomic revision of the bivittatus section.

Nearctic species of Lasiopogon Loew comprising the bivittatus section (the bivittatus group sensu Cannings 2002) are revised, with the description of 13 new species, elevation of one subspecies to species, and redescriptions of 13 previously described taxa. An updated key to western Nearctic Lasiopogon adults is provided, as are notes on taxonomy, distribution, phylogeny, and ecology. A Bayesian species tree for 67 species of Lasiopogon is estimated from one mitochondrial (COI) and three nuclear protein-coding loci (AATS, PEPCK, Wg), and compared to a previously published morphology-based phylogeny. The following new species of Lasiopogon are described (assigned to the bivittatus section except as noted): L. anaphlecter sp. nov., L. apoecus sp. nov., L. asilomar sp. nov., L. bitumineus sp. nov., L. canningsi sp. nov., L. condylophorus sp. nov., L. esau sp. nov., L. karli sp. nov. (assigned to cinereus group of opaculus section), L. nelsoni sp. nov., L. odontotus sp. nov., L. sierra sp. nov., L. tumulicola sp. nov., L. wilcoxi sp. nov.; L. puyallupi Cole Wilcox 1938 stat. nov. is elevated from subspecies; and the following previously described species are considered valid: L. actius Melander 1923, L. albidus Cole Wilcox 1938, L. arenicola (Osten Sacken 1877), L. bivittatus Loew 1866, L. californicus Cole Wilcox 1938, L. dimicki Cole Wilcox 1938, L. drabicolum Cole 1916, L. gabrieli Cole Wilcox 1938, L. littoris Cole 1924, L. ripicola Melander 1923, L. willametti Cole Wilcox 1938, L. zonatus Cole Wilcox 1938. The species L. martinensis Cole Wilcox 1938 is considered valid but transferred to the tetragrammus group of the opaculus section.

Comparative Decellularization and Recellularization of Wild-Type and Alpha 1,3 Galactosyltransferase Knockout Pig Lungs: A Model for Ex Vivo Xenogeneic Lung Bioengineering and Transplantation.

BACKGROUND: A novel potential approach for lung transplantation could be to utilize xenogeneic decellularized pig lung scaffolds that are recellularized with human lung cells. However, pig tissues express several immunogenic proteins, notably galactosylated cell surface glycoproteins resulting from alpha 1,3 galactosyltransferase (α-gal) activity, that could conceivably prevent effective use. Use of lungs from α-gal knock out (α-gal KO) pigs presents a potential alternative and thus comparative de- and recellularization of wild-type and α-gal KO pig lungs was assessed. METHODS: Decellularized lungs were compared by histologic, immunohistochemical, and mass spectrometric techniques. Recellularization was assessed following compartmental inoculation of human lung bronchial epithelial cells, human lung fibroblasts, human bone marrow-derived mesenchymal stromal cells (all via airway inoculation), and human pulmonary vascular endothelial cells (CBF) (vascular inoculation). RESULTS: No obvious differences in histologic structure was observed but an approximate 25% difference in retention of residual proteins was determined between decellularized wild-type and α-gal KO pig lungs, including retention of α-galactosylated epitopes in acellular wild-type pig lungs. However, robust initial recellularization and subsequent growth and proliferation was observed for all cell types with no obvious differences between cells seeded into wild-type versus α-gal KO lungs. CONCLUSION: These proof of concept studies demonstrate that decellularized wild-type and α-gal KO pig lungs can be comparably decellularized and comparably support initial growth of human lung cells, despite some differences in retained proteins. α-Gal KO pig lungs are a suitable platform for further studies of xenogeneic lung regeneration.

Amatoxin and phallotoxin concentration in amanita bisporigera spores.

Even though amatoxins and phallotoxins have been well characterized in basidiocarps of Amanita species, to our knowledge no report of these toxins in spores of Amanitas has been published. Reversed phase HPLC was used to determine non-zero concentrations of alpha-amanitin (0.30 mg/g), and phallacidin (0.02 mg/g) in spores taken from white Amanita sect. phalloideae species. We did not find significant amounts of phalloidin in Amanita spores. We also report concentrations of these toxins from pileus and stipe tissues that are similar to previously reported values, lending support to the hypothesis that toxin concentrations in spores are much less than in other basidiocarp tissues.