Arterial Elastin as Seen with Scanning Electron Microscopy: A Review

All large arteries contain elastin, collagen, and muscle which can be seen with light microscopy and transmission electron microscopy. Elastin forms an internal elastic lamina (IEL) in all arteries, but also forms multiple fenestrated sheets in the media of the aorta and other large arteries. The fenestrations in the media are larger than those in the IEL. The adventitial elastin is more fibrous and often contains tubular elastin surrounding vasa vasorum when prepared by removing all non-elastin by placing the aorta in 0.1 N NaOH at 70-75°C for five hours. The fenestrations are larger near branches and in an experimentally created poststenotic dilatation. Atherosclerosis appears associated with both new elastin formation in early atherosclerosis and elastolysis in late disease

Scanning Electron Microscopy of Early Atherosclerosis in Rabbits Using Aortic Casts

Our research involves measuring the size and location of atherosclerotic lesions on the intimal surface of arteries. To this end we have developed a new method in which scanning electron micrographs of vascular casts with impressions of these lesions on their surface are used. This method is compared with light microscopy and scanning electron microscopy of tissue with lesions. All three methods are found to detect lesions equally well. We also examine the cellular elements in the lesion to determine how the images are formed

Quantitative Measurement from Vascular Casts

A review of quantitative measurements show casting materials shrink from 0.2 - 20% and have viscosities ranging from 1.4 - 100,000 centipoise. Blood vessels have highly variable mechanical properties. Some microvessels are very stiff having little change in dimensions with pressure. Larger vessels generally change diameter significantly but show highly variable changes in length with pressure. Perfusion fixation does not fix the dimensions of blood vessels. Dog carotid arteries well fixed with glutaraldehyde at physiologic dimensions retain ≈20% of their elastic recoil circumferentially and ≈30% longitudinally. We recommend vascular casting as a method of accurately measuring the vasculature if care is taken to use low shrinkage casting resins and maintain physiologic transmural pressures for the duration of any casting procedure, even if prefixation is used. We measured a ≈10% error in our method of measuring both the size and location of periorificial atherosclerotic lesions from aortic casts. Little is known about how vascular smooth muscle tone changes during casting

Loss-of-Function Mutations in PTPN11 Cause Metachondromatosis, but Not Ollier Disease or Maffucci Syndrome

Metachondromatosis (MC) is a rare, autosomal dominant, incompletely penetrant combined exostosis and enchondromatosis tumor syndrome. MC is clinically distinct from other multiple exostosis or multiple enchondromatosis syndromes and is unlinked to EXT1 and EXT2, the genes responsible for autosomal dominant multiple osteochondromas (MO). To identify a gene for MC, we performed linkage analysis with high-density SNP arrays in a single family, used a targeted array to capture exons and promoter sequences from the linked interval in 16 participants from 11 MC families, and sequenced the captured DNA using high-throughput parallel sequencing technologies. DNA capture and parallel sequencing identified heterozygous putative loss-of-function mutations in PTPN11 in 4 of the 11 families. Sanger sequence analysis of PTPN11 coding regions in a total of 17 MC families identified mutations in 10 of them (5 frameshift, 2 nonsense, and 3 splice-site mutations). Copy number analysis of sequencing reads from a second targeted capture that included the entire PTPN11 gene identified an additional family with a 15 kb deletion spanning exon 7 of PTPN11. Microdissected MC lesions from two patients with PTPN11 mutations demonstrated loss-of-heterozygosity for the wild-type allele. We next sequenced PTPN11 in DNA samples from 54 patients with the multiple enchondromatosis disorders Ollier disease or Maffucci syndrome, but found no coding sequence PTPN11 mutations. We conclude that heterozygous loss-of-function mutations in PTPN11 are a frequent cause of MC, that lesions in patients with MC appear to arise following a “second hit,” that MC may be locus heterogeneous since 1 familial and 5 sporadically occurring cases lacked obvious disease-causing PTPN11 mutations, and that PTPN11 mutations are not a common cause of Ollier disease or Maffucci syndrome