Annotation of putative circadian rhythm-associated genes in Diaphorina citri (Hemiptera: Liviidae).

The circadian rhythm involves multiple genes that generate an internal molecular clock, allowing organisms to anticipate environmental conditions produced by the Earth's rotation on its axis. Here, we present the results of the manual curation of 27 genes that are associated with circadian rhythm in the genome of Diaphorina citri, the Asian citrus psyllid. This insect is the vector for the bacterial pathogen Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus greening disease (Huanglongbing). This disease severely affects citrus industries and has drastically decreased crop yields worldwide. Based on cry1 and cry2 identified in the psyllid genome, D. citri likely possesses a circadian model similar to the lepidopteran butterfly, Danaus plexippus. Manual annotation will improve the quality of circadian rhythm gene models, allowing the future development of molecular therapeutics, such as RNA interference or antisense technologies, to target these genes to disrupt the psyllid biology.

Tea tree oil extract causes mitochondrial superoxide production and apoptosis as an anticancer agent, promoting tumor infiltrating neutrophils cytotoxic for breast cancer to induce tumor regression.

The antitumor activity of the tea tree oil (TTO) derived product, Melaleuca Alternifolia Concentrate (MAC) was characterized mechanistically at the molecular and cellular level. MAC was analyzed for its anticancer activity against human prostate (LNCaP) and breast (MCF-7) cancer cell lines growing in vitro. MAC (0.02-0.06% v/v) dose-dependently induced the intrinsic (mitochondrial) apoptotic pathway in both the LNCaP and MCF-7 cell lines, involving increased mitochondrial superoxide production, loss of mitochondrial membrane potential (MMP), caspase 3/7 activation, as well as the presence of TUNEL+ and cleaved-PARP+ cell populations. At concentrations of 0.01-0.04% v/v, MAC caused cell cycle arrest in the G0/1-phase, as well as autophagy. The in vivo anticancer actions of MAC were examined as a treatment in the FVB/N c-Neu murine model for spontaneously arising breast cancers. Intratumoral MAC injections (1-4% v/v) significantly suppressed tumor progression in a dose-dependent manner and was associated with greater levels of tumor infiltrating neutrophils exhibiting anticancer cytotoxic activity. Induction of breast cancer cell death by MAC via the mitochondrial apoptotic pathway was also replicated occurring in tumors treated in vivo. In conclusion, our data highlights the potential for the Melaleuca-derived MAC product inducing anticancer neutrophil influx, supporting its application as a novel therapeutic agent.

Utilizing a chromosomal-length genome assembly to annotate the Wnt signaling pathway in the Asian citrus psyllid, Diaphorina citri.

The Asian citrus psyllid, Diaphorina citri, is an insect vector that transmits Candidatus Liberibacter asiaticus, the causal agent of the Huanglongbing (HLB), or citrus greening disease. This disease has devastated Florida's citrus industry, and threatens California's industry as well as other citrus producing regions around the world. To find novel solutions to the disease, a better understanding of the vector is needed. The D. citri genome has been used to identify and characterize genes involved in Wnt signaling pathways. Wnt signaling is utilized for many important biological processes in metazoans, such as patterning and tissue generation. Curation based on RNA sequencing data and sequence homology confirms 24 Wnt signaling genes within the D. citri genome, including homologs for beta-catenin, Frizzled receptors, and seven Wnt-ligands. Through phylogenetic analysis, we classify D. citri Wnt ligands as Wg/Wnt1, Wnt5, Wnt6, Wnt7, Wnt10, Wnt11, and WntA. The D. citri version 3.0 genome with chromosomal length scaffolds reveals a conserved Wnt1-Wnt6-Wnt10 gene cluster with a gene configuration like that in Drosophila melanogaster. These findings provide greater insight into the evolutionary history of D. citri and Wnt signaling in this important hemipteran vector. Manual annotation was essential for identifying high quality gene models. These gene models can be used to develop molecular systems, such as CRISPR and RNAi, which target and control psyllid populations to manage the spread of HLB. Manual annotation of Wnt signaling pathways was done as part of a collaborative community annotation project.

Medicare bundled payment program: meaningful risk and limited reward.

CMS's Bundled Payments for Care Improvement initiative is accepting applications from healthcare provider organizations that would like to participate in the one of four alternative payment models under the initiative. Three of these models-the All MS-DRG Hospital Model, the Global Hospitalization Model, and the Global Episode Model-target care delivered in an acute care setting. Hospitals contemplating participation in one of the models should weigh the benefits and risks of participation; it is likely the significant risk and limited financial benefit associated with the program will dissuade most providers from participating inthe initiative.

Managing the risks of accountable care.

Hospitals and health systems should assess the risks of participation in Medicare's Shared Savings Program before deciding whether to participate: Start-up costs will likely be prohibitive. The ACO will lack key tools needed to drive down unnecessary medical costs. Participating providers would have to accept significant downside risk for costs over which they have no control. ACOs generating significant savings in a program year would still face numerous preconditions to-and limitations on-payment.

High stakes: avoiding risks associated with vendor discounts.

The federal government has long been concerned about several types of discount arrangements, some of which are increasing in sophistication, including: Swapping. Bundling. Tiered discounts. Free use of high-end equipment in return for protocols or data.

The return of gainsharing.

Hospitals may participate in gainsharing arrangements, but those arrangements should include safeguards that will minimize regulatory exposure. Gainsharing provides a powerful means by which hospitals and physicians can reconcile their conflicting economic interest while promoting quality of care. Hospitals are challenged to structure gainsharing arrangements so they maximize productive hospital-physician collaboration while minimizing the OIG's compliance concerns.

Stark realities for hospitals.

As phase II of the Stark law becomes a reality, hospitals will find that the new physician self-referral regulations fall into three categories: the helpful, the unhelpful, and the marginal. Providers need to understand the principal changes in each category and modify their compliance efforts when compensating physicians.

Gainsharing: a cost-reduction strategy that may be back.

The HHS Office of Inspector General (OIG) seemingly has reversed its formerly unwavering prohibition on gainsharing arrangements. In Advisory Opinion 01-1, the OIG approved an arrangement between a hospital and a physician group targeted at reducing operating expenses as long as they adopted appropriate safeguards to protect against Federal healthcare-program fraud and abuse. With similar approval, hospitals could take advantage of this opportunity to enter into their own gainsharing arrangements with physicians. There are, however, limitations that should be considered when structuring any preliminary gainsharing initiatives. Several Federal laws already exist that define and restrict gainsharing arrangements.