Mitochondrial DNA (mtDNA) analysis of highly degraded skeletal remains is often used for forensic identification due largely to the high genome copy number per cell. Literature from the “ancient DNA” field has shown that highly degraded samples contain populations of intact DNA molecules that are severely restricted in size (1–4). Hand et al. have demonstrated the targeting and preferential amplification of authentic human DNA sequences with small amplicon products of 150 bp or less (1,2). Given this understanding of ancient DNA preservation and amplification, we report an improved approach to forensic mtDNA analysis of hypervariable regions 1 and 2 (HV1/HV2) in highly degraded specimens. This “mini-primer set” (MPS) amplification strategy consists of four overlapping products that span each of the HV regions and range from 126 to 170 bp, with an average size of 141 bp. For this study, 11 extracts representing a range of sample quality were prepared from nonprobative forensic specimens. We demonstrate a significant increase in MPS amplification success when compared to testing methods using ∼250 bp amplicons. Further, 16 of 17 independent amplifications previously “unreported” due to mixed sequences provided potentially reportable sequence data from a single, authentic template with MPS testing.

Fragment-based drug discovery (FBDD) is contingent on the development of analytical methods to identify weak protein-fragment noncovalent interactions. Herein we have combined an underutilized fragment screening method, native state mass spectrometry, together with two proven and popular fragment screening methods, surface plasmon resonance and X-ray crystallography, in a fragment screening campaign against human carbonic anhydrase II (CA II). In an initial fragment screen against a 720-member fragment library (the "CSIRO Fragment Library") seven CA II binding fragments, including a selection of nonclassical CA II binding chemotypes, were identified. A further 70 compounds that comprised the initial hit chemotypes were subsequently sourced from the full CSIRO compound collection and screened. The fragment results were extremely well correlated across the three methods. Our findings demonstrate that there is a tremendous opportunity to apply native state mass spectrometry as a complementary fragment screening method to accelerate drug discovery.

Diphenyliodine(III) triflate is able to transfer Ph + to Pd(II) and Pt(II) with cleavage of a phenyl-iodine bond and formation of metal(IV) species, leading to the first identified transfer of Ph + to Pd(II) from an aryl-halogen bond, and, for platinum, a methodology providing a facile route to dimethyl(aryl)platinum(IV) and 1,4-arenediyl-bridged Pt(IV) species and the first archetypal triarylplatinum(IV) complex. Thus, [IPh 2 ][OTf] reacts with PtMe 2 (bpy) (bpy ) 2,2′-bipyridine) at -50°C to form iodobenzene and the Pt(IV) complex trans-Pt IV (OTf)Me 2 -Ph(N∼N) (1b) (Ph trans to OTf), and on addition of NaI, the species PtIMe 2 Ph(bpy) (2a (Ph cis to I) and 2b (Ph trans to I) in 2:1 ratio) may be isolated at -20°C. Similarly, metalla-(II)cyclopentane complexes M(C 4 H 8 )(bpy) react with [IPh 2 ][OTf] to form trans-Pt(OTf)(C 4 H 8 )-Ph(bpy) (3b) and a 1:1 ratio of cis-(4a) and trans-Pd(OTf)(C 4 H 8 )Ph(bpy) (4b); addition of halide ion gives trans-PtI(C 4 H 8 )Ph(bpy) (5b) and a 1:3 ratio of cis and trans isomers for PdI(C 4 H 8 )Ph(bpy) (6a, 6b) and PdCl(C 4 H 8 )Ph(bpy) (7a, 7b). Complex 5b isomerizes to form a 2:1 mixture of cis-PtI(C 4 H 8 )Ph(bpy) (5a) and 5b at ambient temperature in acetone. Dimethyl(2,2′-bipyridine)palladium(II) reacts with [IPh 2 ][OTf] to form Pd(OTf)Me 2 Ph(bpy), followed by transfer of a methyl group from Pd(IV) to Pd(II), to form trimethylpalladium-(IV) species. Dimethyl(2,2′-bipyridine)platinum(II) reacts with [IPh(C 6 H 4 -4-I)][OTf], followed by addition of sodium iodide, to form a 1:1 mixture of trans-PtIMe 2 Ph(bpy) (2b) and transPtIMe 2 (C 6 H 4 -4-I)(bpy) (8b), and with [IPh(C 6 H 4 -4-IPh)][OTf] 2 to form the 1,4-arenediyl complex trans-1,4-{PtIMe 2 (bpy)} 2 C 6 H 4 (9b). Diphenyl{di(tert-butyl)-2,2′-bipyridine}platinum-(II) reacts with [IPh 2 ][OTf] at 25°C over 2 days to form the triphenylplatinum(IV) complex Pt(OTf)Ph 3 (Bu t 2 bpy) (10), and addition of iodide ion results in isolation of PtIPh 3 (Bu t 2 bpy) (11). Structural studies of trans-PtIMe 2 Ph(bpy) (2b) and trans-Pt(C 4 H 8 )Ph(bpy) (5b) reveal distorted octahedral geometry and the fac-PtC 3 configuration expected for all of the metal-(IV) complexes. The compound [IPh(C 6 H 4 -4-I)][OTf] has two sets of cation-anion pairs with a complex array of weak interactions, and the cations have C-I-C angles close to 90°.

The total synthesis of the spiropiperidine alkaloid (-)-perhydrohistrionicotoxin (perhydro-HTX) 2 has been accomplished on a gram scale by employing both conventional batch chemistry as well as microreactor techniques. (S)-(-)-6-Pentyltetrahydro-pyran-2-one 8 underwent nucleophilic ring opening to afford the alcohol 10, which was elaborated to the nitrone 13. Protection of the nitrone as the 1,3-adduct of styrene and side-chain extension to the unsaturated nitrile afforded a precursor 17, which underwent dipolar cycloreversion and 1,3-dipolar cycloaddition to give the core spirocyclic precursor 18 that was converted into perhydro-HTX 2. The principal steps to the spirocycle 18 have successfully been transferred into flow mode by using different types of microreactors and in a telescoped fashion, allowing for a more rapid access to the histrionicotoxins and their analogues by continuous processing.