Apoptosis and necrosis, two major forms of cell death, can be distinguished morphologically and biochemically. Internucleosomal DNA fragmentation (INDF) is a biochemical hallmark of apoptosis, and caspase-activated DNase (CAD), also known as DNA fragmentation factor 40 kDa (DFF40), is one of the major effector endonucleases. DNase γ, a Mg2+/Ca2+-dependent endonuclease, is also known to generate INDF but its role among other apoptosis-associated endonucleases in cell death is unclear. Here we show that (i) INDF occurs even during necrosis in cell lines, primary cells, and in tissues of mice in vivo, and (ii) DNase γ, but not CAD, is the effector endonuclease for INDF in cells undergoing necrosis. These results document a previously unappreciated role for INDF in necrosis and define its molecular basis.

Major depression is often a relapsing disorder. It is therefore important to start its treatment with therapies that maximize the chance of not only getting the patients well but also keeping them well. We examined the associations between initial treatments and sustained response by conducting a network meta-analysis of randomized controlled trials (RCTs) in which adult patients with major depression were randomized to acute treatment with a psychotherapy (PSY), a protocolized antidepressant pharmacotherapy (PHA), their combination (COM), standard treatment in primary or secondary care (STD), or pill placebo, and were then followed up through a maintenance phase. By design, acute phase treatment could be continued into the maintenance phase, switched to another treatment or followed by discretionary treatment. We included 81 RCTs, with 13,722 participants. Sustained response was defined as responding to the acute treatment and subsequently having no depressive relapse through the maintenance phase (mean duration: 42.2±16.2 weeks, range 24-104 weeks). We extracted the data reported at the time point closest to 12 months. COM resulted in more sustained response than PHA, both when these treatments were continued into the maintenance phase (OR=2.

Exiguobacterium sp. F42 was screened as a producer of an enzyme catalyzing the NADPH-dependent stereoselective reduction of ethyl 3-oxo-3-(2-thienyl)propanoate (KEES) to ethyl (S)-3-hydroxy-3-(2-thienyl)propanoate ((S)-HEES). (S)-HEES is a key intermediate for the synthesis of (S)-duloxetine, a potent inhibitor of the serotonin and norepinephrine uptake carriers. The responsible enzyme (KEES reductase) was partially purified, and the gene encoding KEES reductase was cloned and sequenced via an inverse PCR approach. Sequence analysis of the gene for KEES reductase revealed that the enzyme was a member of the short chain dehydrogenase/reductase family. The probable NADPH-interacting site and 3 catalytic residues (Ser-Tyr-Lys) were fully conserved. The gene was highly expressed in Escherichia coli, and the gene product was purified to homogeneity from the recombinant E. coli by simpler procedures than from the original host. The molecular mass of the purified enzyme was 27,500 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 55,000 as determined by gel filtration chromatography. Our results show that this enzyme can be used for the practical production of (S)-HEES.

This paper reports the temporal structures of the relationships between 24-hour heart rate variability (HRV) and physical activity (PA) in clinical big data of simultaneously recorded electrocardiograms (ECG) and actigraphic data in 21,682 male and 29,446 female patients (age, 20-100 yr). We found that SDNN (standard deviation of 24-hour normal-to-normal R-R interval [NN interval] of ECG) and PA during daytime, evening, and nighttime decreased with age. In regression models adjusted for the effect of age, daytime PA explained 7% of inter-individual variance in SDNN, while evening and early morning PA explained only <0.6% and nighttime PA negatively contributed to SDNN. Daytime but not nighttime PA is positively associated with increased 24-hour HRV.

DNA fragmentation is a biochemical hallmark of apoptosis. Several endonucleases, including CAD/DFF40 and endonuclease G, are implicated in DNA fragmentation. DNase γ has also been considered to be one of the enzymes involved, but its role in relation to CAD/DFF40 in apoptosis has not been fully elucidated. Here, we distinguished between DNase γ-dependent and CAD/ DFF40-dependent DNA fragmentations. We found that DNase γ activities appeared in the late apoptotic phase and accelerated DNA fragmentation. Thus, even if the apoptotic DNA fragmentation is initiated by CAD/DFF40, DNase γ is required for the more complete digestion of the genomic DNA in dying cells.Apoptosis, or programmed cell death, eliminates unwanted cells during development, homeostasis, and immune responses (11,20). It is characterized by changes in cell morphology, including the formation of membrane-associated apoptotic bodies, membrane blebbing, nuclear breakdown, and chromatin condensation. At the molecular level, these changes are accompanied by the cleavage of several proteins and DNA fragmentation. DNA fragmentation is the final step in the disposal of the genomic DNA in cells undergoing apoptosis. The fragmentation of the genomic DNA in apoptotic cells occurs in two steps: the initial macrofragmentation (fragments ≧ 50 kb, a size consistent with that of a chromatin loop domain) and the subsequent internucleosomal DNA fragmentation, which is often observed as DNA laddering (12). The DNA fragmentation during apoptosis is not a passive process but rather an active process that participates in the enhancement of the apoptotic cascade in response to proapoptotic stimulations (2). However, the mechanism of the DNA fragmentation in apoptotic cells remains to be clarified. Although several endonucleases have been implicated in this process, their specific roles are still under debate. The DNA fragmentation factor (DFF) is a wellcharacterized apoptotic endonuclease (9). DFF is a heterodimer composed of a 40-kDa molecule and a 45-kDa molecule (DFF40 and DFF45, respectively).