Recently, the JAK2 V617F mutation was found in patients with myeloproliferative disorders (MPDs), including most with polycythemia vera (PV). The mutant JAK2 has increased kinase activity, and it was shown to be pathogenic in mouse models. Herein, we analyzed blood samples randomly collected from a clinical laboratory. Surprisingly, as many as 37 samples from a total of 3935 were found positive for the JAK2 mutation. However, only one of these samples had blood test results indicative for probable PV, but several had nonhematologic diseases. On average, samples with the mutation had normal red cell counts but significantly higher white blood cell and platelet counts, although most were within the normal range. The data suggest that the JAK2 V617F mutation is apparently much more common than MPDs. Its occurrence may be a prelude to full blood cell abnormalities and other diseases, but it cannot by itself diagnose MPDs. IntroductionProtein tyrosine kinases are central regulators of signaling pathways and are attractive therapeutic targets. [1][2][3] Recently, several groups identified a recurrent somatic activating mutation in the JAK2 tyrosine kinase in polycythemia vera (PV), essential thrombocythemia (ET), and idiopathic myelofibrosis. 4-8 Infrequent occurrence of this mutation has been also reported in chronic myelomonocytic leukemia, atypical myeloproliferative disorders (MPDs), myelodysplastic syndrome, systemic mastocytosis, chronic neutrophilic leukemia, and acute myeloid leukemia. [9][10][11][12][13] This mutation results in a valine to phenylalanine substitution within the pseudokinase domain of JAK2. The mutant JAK2 possesses enhanced tyrosine kinase activity and causes a PV-like phenotype in mouse bone marrow transplantation models. 6,14,15 However, it remains unclear whether the JAK2 V617F mutation is solely responsible for these diseases and why it is associated with such a wide spectrum of phenotypes. Previous investigations were carried out to identify JAK2 V617F in samples from specific diseases. In this study, we analyzed blood samples randomly collected from a clinical laboratory. Materials and methods Blood samplesDeidentified, consecutive peripheral blood samples from donors (all Chinese) of any age, sex, and disease were randomly collected from the clinical laboratory of China Japan Union Hospital, Changchun, China, over a period of about 6 months. Institutional Review Board approvals were obtained from both Jilin University and the University of Oklahoma Health Science Center. PCR amplificationGenomic DNA was extracted from the whole blood. A genomic DNA fragment containing the exon 14 of the JAK2 gene which bears the V617F mutation site was amplified by polymerase chain reaction (PCR) with primers P1 and P1r ( Figure 1A). The PCR was run with Taq DNA polymerase or Phusion polymerase (for confirmation of the positive samples) for 35 cycles with 94°C for 20 seconds, 60°C for 20 seconds, and 72°C for 30 seconds. For allele-specific PCR, 0.2 L of the initial PCR product was used for further PC...

Tandem reactions of Pd‐catalyzed cross‐coupling of 3‐(2‐isocyanoethyl)indoles with diazoacetates and subsequent spirocyclization/Mannich‐type reaction have been developed to assemble polycyclic spiroindoline skeletons. Formation of spiroindolenines has been proven as the crucial step for the following Mannich‐type cyclization reaction. Accordingly, a novel approach on chiral phosphoric acid catalyzed Mannich‐type cyclization toward the formation of diastereomerically and enantiomerically enriched pentacyclic spiroindolines has been established. Moreover, the products of the reaction are versatile building blocks in synthetic chemistry, as demonstrated by the synthesis of the key framework of aspidosperma and kopsia alkaloids.

The development of catalytic enantioselective isocyanide‐based reactions is currently of great interest because the resulting products are valuable in organic synthesis, pharmacological chemistry, and materials science. This review assembles and comprehensively summarizes the recent achievements in this rapidly growing area according to the reaction types. Special attention is paid to the advantages, limitations, possible mechanisms, and synthetic applications of each reaction. In addition, a personal outlook on the opportunities for further exploration is given at the end.

Selective ion-pair binding of CaX2 (X = Br-, I-) was realized by a tritopic receptor containing homoditopic anion-π binding sites and a pentaethylene glycol site. The receptor formed an unusual solvent-separated ion-pair complex with CaI2 and a contact one with CaBr2, and can be applied to selectively dissolve calcium halides in organic solvents.