Ionotropic Receptor-dependent cool cells control the transition of temperature preference in Drosophila larvae

Temperature sensation guides animals to avoid temperature extremes and to seek their optimal temperatures. The larval stage of Drosophila development has a dramatic effect on temperature preference. While early-stage Drosophila larvae pursue a warm temperature, late-stage larvae seek a significantly lower temperature. Previous studies suggest that this transition depends on multiple rhodopsins at the late larval stage. Here, we show that early-stage larvae, in which dorsal organ cool cells (DOCCs) are functionally blocked, exhibit similar cool preference to that of wild type late-stage larvae. The molecular thermoreceptors in DOCCs are formed by three members of the Ionotropic Receptor (IR) family, IR21a, IR93a, and IR25a. Early-stage larvae of each Ir mutant pursue a cool temperature, similar to that of wild type late-stage larvae. At the late larval stage, DOCCs express decreased IR proteins and exhibit reduced cool responses. Importantly, late-stage larvae that overexpress IR21a, IR93a, and IR25a in DOCCs exhibit similar warm preference to that of wild type early-stage larvae. These data suggest that IR21a, IR93a, and IR25a in DOCCs navigate early-stage larvae to avoid cool temperatures and the reduction of these IR proteins in DOCCs results in animals remaining in cool regions during the late larval stage. Together with previous studies, we conclude that multiple temperature-sensing systems are regulated for the transition of temperature preference in fruit fly larvae.

Download Full-text

Endoplasmic Reticulum Stress Is a Target for Therapy in Paroxysmal Nocturnal Hemoglobinuria.

Blood ◽

10.1182/blood.v114.22.160.160 ◽

2009 ◽

Vol 114 (22) ◽

pp. 160-160

Author(s):

Regis Peffault de Latour ◽

Valeria Visconte ◽

Keyvan Keyvanfar ◽

Olga Nunez ◽

Marie Desierto ◽

...

Keyword(s):

Flow Cytometry ◽

Cell Line ◽

Late Stage ◽

Early Stage ◽

Proteasome Inhibitors ◽

Wild Type ◽

Deficient Cell ◽

Cd34 Cells ◽

Wild Type Counterpart ◽

Apoptosis Assay

Abstract Abstract 160 Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by hemolytic anemia, bone marrow failure and venous thromboembolism. The disease is caused by a somatic mutation of the X-linked gene PIG-A, encoding a key enzyme responsible for the biosynthesis of the glycosylphosphatidylinositol anchored proteins (GPI-APs). Unfolded GPI-APs are translated and degraded intracellularly in the endoplasmic reticulum (ER) via the proteasome machinery. We hypothesized that the accumulation of misfolded proteins in mutant PNH cells activates the unfolded protein response (UPR) signaling cascade, which physiologically maintains the quality of newly synthesized proteins. UPR could thus represent a new target for therapy in PNH. We first assessed the sensitivity of GPI-AP- deficient K562 cells to the proteasome inhibitor PS-341 (Velcade‘) by flow cytometry after staining for annexin V and propidium iodide (PI). After 24 hour exposure to PS-341, cytotoxicity was significantly higher in the K562 GPI-AP-deficient cell line compared to their wild type counterpart at all dose tested (0 to 200nM). At 50nM, the early stage apoptosis was 21±2% in the K562 GPI-AP-deficient cell line versus 9.3±2% for their wild type counterpart (p=0.013) while the late stage apoptosis was 38±2.3% versus 22±2.3% (p<10−3), respectively. Similar results were obtained in immortalized lymphoblastoid cells derived from a PNH patient after PS-341 exposure. We reproduced those results with 2 other proteasome inhibitors (MG132 and Lactacystine) confirming the fundamental role of the proteasome in PNH. By immunoblot, K562 GPI-AP-deficient cell line exhibited more expression of chaperone proteins (CHOP and BiP/GRP78) compared to wild type cells after PS-341 exposure; evidence of the UPR is initiated by accumulation of unfolded proteins. UPR activation was confirmed by the detection of XBP1 splicing form in K562 GPI-AP-deficient cells by polymerase chain reaction (PCR). Moreover, PS-341 promoted pro-apoptotic/terminal UPR gene expression in K562 GPI-APs deficient cells as illustrated by the increase expression of an UPR specific pro-apoptotic protein, NOXA. We observed a concomitant arrest in the G2 phase of the cell cycle, as detected by flow cytometry after staining with PI. We employed a mouse model bearing a conditional Pig-a gene deletion in hematopoietic cells. When the animals were treated with PS-341 (15 ug/day once a week, intraperitoneally), there was a significantly higher proportion of apoptotic GPI-AP-deficient B cells compared to normal B cells in early stage apoptosis (p=0.01) and a slight increase of in the late stage apoptosis (p=0.13). PS-341 induced apoptosis also in bone marrow samples from two patients with classical and aplastic PNH as assessed by flow cytometry, based on CD59 and CD55 expression in the CD34+ population (Figure 1). GPI-AP-CD34+ deficient cells were more sensitive to PS-341 at the early stage of apoptosis (at 50 nM, 14% vs 4.6%), as well as late stage (at 200nM: 22% vs 7.2%). In conclusion, we demonstrate that GPI-AP-deficient cells are more susceptible to proteasome inhibition through the activation of the proapoptotic/terminal UPR, which leads to specific apoptosis in these cells. The sensitivity of PNH cells in vivo and in vitro to proteasome inhibitors might allow for the development of therapeutic strategies specifically directed to target one or several components of the UPR in PNH.Figure 1:Representative apoptosis assay in CD34+ cells according to GPI expression after 24 hours exposition to PS-341Figure 1:. Representative apoptosis assay in CD34+ cells according to GPI expression after 24 hours exposition to PS-341 Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Making the Grade during Pandemic: Early-stage and Late-stage Provisional Institutions

Sociological Perspectives ◽

10.1177/07311214211028616 ◽

2021 ◽

pp. 073112142110286

Author(s):

Alexander B. Kinney ◽

Nicholas J. Rowland

Keyword(s):

Late Stage ◽

Early Stage ◽

State University ◽

Institutional Work ◽

Traditional Grading ◽

Grading Policy ◽

Temporary Solution ◽

Rural State ◽

Empirical Implications ◽

Common Understanding

This is an article that draws on the institutional work literature about provisional institutions. To date, nearly every U.S. sector has been impacted by COVID-19. To sustain their core missions, highly institutionalized organizations such as universities have had to rethink foundational structures and policies. Using a historical ethnographic approach to investigate records from faculty senate deliberations at “Rural State University” (RSU), the authors examine the implementation of a temporary grading policy to supplement traditional, qualitative grades spring 2020 during the outbreak. The authors find that RSU implemented a temporary, supplemental grading policy as a provisional institution to momentarily supersede traditional grading as a means to—as soon as possible—return to it. This finding contrasts with the common understanding that provisional institutions operate primarily as a temporary solution to a social problem that leads to more stable and enduring, ostensibly nonprovisional institutions. The temporary grading policy, the authors argue, constitutes a “late-stage” provisional institution and, with this new lens, subsequently characterize the more commonplace understanding of provisional institutions as “early-stage.” This contribution has theoretical implications for studies of institutions and empirical implications for research on shared governance and disruption in higher education.

Download Full-text

Health Insurance Status as a Predictor of Mode of Colon Cancer Detection but Not Stage at Diagnosis: Implications for Early Detection

Public Health Reports ◽

10.1177/0033354921999173 ◽

2021 ◽

pp. 003335492199917

Author(s):

Lindsey A. Jones ◽

Katherine C. Brewer ◽

Leslie R. Carnahan ◽

Jennifer A. Parsons ◽

Blase N. Polite ◽

...

Keyword(s):

Colon Cancer ◽

Health Insurance ◽

Early Detection ◽

Late Stage ◽

Early Stage ◽

Insurance Status ◽

Stage At Diagnosis ◽

Health Insurance Status ◽

Percentage Point Increase ◽

Point Increase

Objective For colon cancer patients, one goal of health insurance is to improve access to screening that leads to early detection, early-stage diagnosis, and polyp removal, all of which results in easier treatment and better outcomes. We examined associations among health insurance status, mode of detection (screen detection vs symptomatic presentation), and stage at diagnosis (early vs late) in a diverse sample of patients recently diagnosed with colon cancer from the Chicago metropolitan area. Methods Data came from the Colon Cancer Patterns of Care in Chicago study of racial and socioeconomic disparities in colon cancer screening, diagnosis, and care. We collected data from the medical records of non-Hispanic Black and non-Hispanic White patients aged ≥50 and diagnosed with colon cancer from October 2010 through January 2014 (N = 348). We used logistic regression with marginal standardization to model associations between health insurance status and study outcomes. Results After adjusting for age, race, sex, and socioeconomic status, being continuously insured 5 years before diagnosis and through diagnosis was associated with a 20 (95% CI, 8-33) percentage-point increase in prevalence of screen detection. Screen detection in turn was associated with a 15 (95% CI, 3-27) percentage-point increase in early-stage diagnosis; however, nearly half (47%; n = 54) of the 114 screen-detected patients were still diagnosed at late stage (stage 3 or 4). Health insurance status was not associated with earlier stage at diagnosis. Conclusions For health insurance to effectively shift stage at diagnosis, stronger associations are needed between health insurance and screening-related detection; between screening-related detection and early stage at diagnosis; or both. Findings also highlight the need to better understand factors contributing to late-stage colon cancer diagnosis despite screen detection.

Download Full-text

EU FP7 research funding for an orphan drug (Orfadin®) and vaccine (Hep C) development: a success and a failure?

Journal of Pharmaceutical Policy and Practice ◽

10.1186/s40545-021-00317-8 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

L. Schmidt ◽

O. Sehic ◽

C. Wild

Keyword(s):

Hepatitis C ◽

Late Stage ◽

Research Funding ◽

Early Stage ◽

Basic Research ◽

Pharmaceutical Products ◽

Clinical Development ◽

Clinical Indication ◽

Market Authorisation ◽

Risk Capital

Abstract Background We considered the extent of the contribution of publicly funded research to the late-stage clinical development of pharmaceuticals and medicinal products, based on the European Commission (EC) FP7 research funding programme. Using two EC FP7-HEALTH case study examples—representing two types of outcomes—we then estimated wider public and charitable research funding contributions. Methods Using the publicly available database of FP7-HEALTH funded projects, we identified awards relating to late-stage clinical development according to the systematic application of inclusion and exclusion criteria, classified them according to product type and clinical indication, and calculated total EC funding amounts. We then identified two case studies representing extreme outcomes: failure to proceed with the product (hepatitis C vaccine) and successful market authorisation (Orfadin® for alkaptonuria). Total public and philanthropic research funding contributions to these products were then estimated using publicly available information on funding. Results 12.3% (120/977) of all EC FP7-HEALTH awards related to the funding of late-stage clinical research, totalling € 686,871,399. Pharmaceutical products and vaccines together accounted for 84% of these late-stage clinical development research awards and 70% of its funding. The hepatitis C vaccine received total European Community (FP7 and its predecessor, EC Framework VI) funding of €13,183,813; total public and charitable research funding for this product development was estimated at € 77,060,102. The industry sponsor does not consider further development of this product viable; this now represents public risk investment. FP7 funding for the late-stage development of Orfadin® for alkaptonuria was so important that the trials it funded formed the basis for market authorisation, but it is not clear whether the price of the treatment (over €20,000 per patient per year) adequately reflects the substantial public funding contribution. Conclusions Public and charitable research funding plays an essential role, not just in early stage basic research, but also in the late-stage clinical development of products prior to market authorisation. In addition, it provides risk capital for failed products. Within this context, we consider further discussions about a public return on investment and its reflection in pricing policies and decisions justified.

Download Full-text

Natural Zeitgebers Under Temperate Conditions Cannot Compensate for the Loss of a Functional Circadian Clock in Timing of a Vital Behavior in Drosophila

Journal of Biological Rhythms ◽

10.1177/0748730421998112 ◽

2021 ◽

pp. 074873042199811

Author(s):

Franziska Ruf ◽

Oliver Mitesser ◽

Simon Tii Mungwa ◽

Melanie Horn ◽

Dirk Rieger ◽

...

Keyword(s):

Molecular Clock ◽

Time Window ◽

Fruit Fly ◽

Diel Activity ◽

Wild Type ◽

Statistical Measures ◽

Clock Mutant ◽

Full Complement ◽

The Right

The adaptive significance of adjusting behavioral activities to the right time of the day seems obvious. Laboratory studies implicated an important role of circadian clocks in behavioral timing and rhythmicity. Yet, recent studies on clock-mutant animals questioned this importance under more naturalistic settings, as various clock mutants showed nearly normal diel activity rhythms under seminatural zeitgeber conditions. We here report evidence that proper timing of eclosion, a vital behavior of the fruit fly Drosophila melanogaster, requires a functional molecular clock under quasi-natural conditions. In contrast to wild-type flies, period01 mutants with a defective molecular clock showed impaired rhythmicity and gating in a temperate environment even in the presence of a full complement of abiotic zeitgebers. Although period01 mutants still eclosed during a certain time window during the day, this time window was much broader and loosely defined, and rhythmicity was lower or lost as classified by various statistical measures. Moreover, peak eclosion time became more susceptible to variable day-to-day changes of light. In contrast, flies with impaired peptidergic interclock signaling ( Pdf01 and han5304 PDF receptor mutants) eclosed mostly rhythmically with normal gate sizes, similar to wild-type controls. Our results suggest that the presence of natural zeitgebers is not sufficient, and a functional molecular clock is required to induce stable temporal eclosion patterns in flies under temperate conditions with considerable day-to-day variation in light intensity and temperature. Temperate zeitgebers are, however, sufficient to functionally rescue a loss of PDF-mediated clock-internal and -output signaling

Download Full-text

Context-Specific Efficacy of Apalutamide Therapy in Preclinical Models of Pten-Deficient Prostate Cancer

Cancers ◽

10.3390/cancers13163975 ◽

2021 ◽

Vol 13 (16) ◽

pp. 3975

Author(s):

Marco A. De Velasco ◽

Yurie Kura ◽

Naomi Ando ◽

Noriko Sako ◽

Eri Banno ◽

...

Keyword(s):

Prostate Cancer ◽

Antitumor Activity ◽

Cancer Cells ◽

Late Stage ◽

Early Stage ◽

Castration Resistant Prostate Cancer ◽

Akt Inhibitor ◽

Molecular Features ◽

Context Specific

Significant improvements with apalutamide, a nonsteroidal antiandrogen used to treat patients suffering from advanced prostate cancer (PCa), have prompted evaluation for additional indications and therapeutic development with other agents; however, persistent androgen receptor (AR) signaling remains problematic. We used autochthonous mouse models of Pten-deficient PCa to examine the context-specific antitumor activity of apalutamide and profile its molecular responses. Overall, apalutamide showed potent antitumor activity in both early-stage and late-stage models of castration-naïve prostate cancer (CNPC). Molecular profiling by Western blot and immunohistochemistry associated persistent surviving cancer cells with upregulated AKT signaling. While apalutamide was ineffective in an early-stage model of castration-resistant prostate cancer (CRPC), it tended to prolong survival in late-stage CRPC. Molecular features associated with surviving cancer cells in CRPC included upregulated aberrant-AR, and phosphorylated S6 and proline-rich Akt substrate of 40 kDa (PRAS40). Strong synergy was observed with the pan-AKT inhibitor GSK690693 and apalutamide in vitro against the CNPC- and CRPC-derived cell lines and tended to improve the antitumor responses in CNPC but not CRPC in vivo. Upregulation of signal transducer and activator of transcription 3 (STAT3) and proviral insertion in murine-1 (PIM-1) were associated with combined apalutamide/GSK690693. Our findings show that apalutamide can attenuate Pten-deficient PCa in a context-specific manner and provides data that can be used to further study and, possibly, develop additional combinations with apalutamide.

Download Full-text

Lung Cancer Inequalities in Stage of Diagnosis in Ontario, Canada

Current Oncology ◽

10.3390/curroncol28030181 ◽

2021 ◽

Vol 28 (3) ◽

pp. 1946-1956

Author(s):

Aisha K. Lofters ◽

Evgenia Gatov ◽

Hong Lu ◽

Nancy N. Baxter ◽

Sara J. T. Guilcher ◽

...

Keyword(s):

Lung Cancer ◽

Relative Risk ◽

Late Stage ◽

Early Stage ◽

Small Cell ◽

Cancer Type ◽

Adjusted Relative Risk ◽

Late Stage Diagnosis ◽

Adjusted Model

Lung cancer is the most common cancer and cause of cancer death in Canada, with approximately 50% of cases diagnosed at stage IV. Sociodemographic inequalities in lung cancer diagnosis have been documented, but it is not known if inequalities exist with respect to immigration status. We used multiple linked health-administrative databases to create a cohort of Ontarians 40–105 years of age who were diagnosed with an incident lung cancer between 1 April 2012 and 31 March 2017. We used modified Poisson regression with robust standard errors to examine the risk of diagnosis at late vs. early stage among immigrants compared to long-term residents. The fully adjusted model included age, sex, neighborhood-area income quintile, number of Aggregated Diagnosis Group (ADG) comorbidities, cancer type, number of prior primary care visits, and continuity of care. Approximately 62% of 38,788 people with an incident lung cancer from 2012 to 2017 were diagnosed at a late stage. Immigrants to the province were no more likely to have a late-stage diagnosis than long-term residents (63.5% vs. 62.0%, relative risk (RR): 1.01 (95% confidence interval (CI): 0.99–1.04), adjusted relative risk (ARR): 1.02 (95% CI: 0.99–1.05)). However, in fully adjusted models, people with more comorbidities were less likely to have a late-stage diagnosis (adjusted relative risk (ARR): 0.82 (95% CI: 0.80–0.84) for those with 10+ vs. 0–5 ADGs). Compared to adenocarcinoma, small cell carcinoma was more likely to be diagnosed at a late stage (ARR: 1.29; 95% CI: 1.27–1.31), and squamous cell (ARR: 0.89; 95% CI: 0.87–0.91) and other lung cancers (ARR: 0.93; 95% CI: 0.91–0.94) were more likely to be diagnosed at an early stage. Men were also slightly more likely to have late-stage diagnosis in the fully adjusted model (ARR: 1.08; 95% CI: 1.05–1.08). Lung cancer in Ontario is a high-fatality cancer that is frequently diagnosed at a late stage. Having fewer comorbidities and being diagnosed with small cell carcinoma was associated with a late-stage diagnosis. The former group may have less health system contact, and the latter group has the lung cancer type most closely associated with smoking. As lung cancer screening programs start to be implemented across Canada, targeted outreach to men and to smokers, increasing awareness about screening, and connecting every Canadian with primary care should be system priorities.

Download Full-text

Characterization of neuroendocrine tumors in heterozygous mutant MENX rats: a novel model of invasive medullary thyroid carcinoma

Endocrine Related Cancer ◽

10.1530/erc-17-0456 ◽

2018 ◽

Vol 25 (2) ◽

pp. 145-162 ◽

Cited By ~ 2

Author(s):

Sara Molatore ◽

Andrea Kügler ◽

Martin Irmler ◽

Tobias Wiedemann ◽

Frauke Neff ◽

...

Keyword(s):

Neuroendocrine Tumors ◽

Early Stage ◽

Pituitary Tumors ◽

Transcriptome Profiling ◽

Thyroid Neoplasms ◽

Recessive Trait ◽

Wild Type ◽

Incurable Cancer ◽

Medullary Thyroid ◽

Adrenal Pheochromocytoma

Rats affected by the MENX syndrome spontaneously develop multiple neuroendocrine tumors (NETs) including adrenal, pituitary and thyroid gland neoplasms. MENX was initially reported to be inherited as a recessive trait and affected rats were found to be homozygous for the predisposingCdkn1bmutation encoding p27. We here report that heterozygous MENX-mutant rats (p27+/mut) develop the same spectrum of NETs seen in the homozygous (p27mut/mut) animals but with slower progression. Consequently, p27+/mut rats have a significantly shorter lifespan compared with their wild-type (p27+/+) littermates. In the tumors of p27+/mut rats, the wild-typeCdkn1ballele is neither lost nor silenced, implying that p27 is haploinsufficient for tumor suppression in this model. Transcriptome profiling of rat adrenal (pheochromocytoma) and pituitary tumors having different p27 dosages revealed a tissue-specific, dose-dependent effect of p27 on gene expression. In p27+/mut rats, thyroid neoplasms progress to invasive and metastatic medullary thyroid carcinomas (MTCs) accompanied by increased calcitonin levels, as in humans. Comparison of expression signatures of late-stage vs early-stage MTCs from p27+/mut rats identified genes potentially involved in tumor aggressiveness. The expression of a subset of these genes was evaluated in human MTCs and found to be associated with aggressive RET-M918T-positive tumors. Altogether, p27 haploinsufficiency in MENX rats uncovered a novel, representative model of invasive and metastatic MTC exploitable for translational studies of this often aggressive and incurable cancer.

Download Full-text

Endothelial dysfunction and arterial pressure regulation during early diabetes in mice: roles for nitric oxide and endothelium-derived hyperpolarizing factor

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00807.2006 ◽

2007 ◽

Vol 293 (2) ◽

pp. R707-R713 ◽

Cited By ~ 21

Author(s):

Sharyn M. Fitzgerald ◽

Barbara K. Kemp-Harper ◽

Helena C. Parkington ◽

Geoffrey A. Head ◽

Roger G. Evans

Keyword(s):

Nitric Oxide ◽

Endothelial Dysfunction ◽

Arterial Pressure ◽

Early Stage ◽

Mesenteric Arteries ◽

No Production ◽

Wild Type ◽

Early Diabetes ◽

Diabetes In Mice ◽

Vehicle Treatment

We determined whether nitric oxide (NO) counters the development of hypertension at the onset of diabetes in mice, whether this is dependent on endothelial NO synthase (eNOS), and whether non-NO endothelium-dependent vasodilator mechanisms are altered in diabetes in mice. Male mice were instrumented for chronic measurement of mean arterial pressure (MAP). In wild-type mice, MAP was greater after 5 wk of Nω-nitro-l-arginine methyl ester (l-NAME; 100 mg·kg−1·day−1 in drinking water; 97 ± 3 mmHg) than after vehicle treatment (88 ± 3 mmHg). MAP was also elevated in eNOS null mice (113 ± 4 mmHg). Seven days after streptozotocin treatment (200 mg/kg iv) MAP was further increased in l-NAME-treated mice (108 ± 5 mmHg) but not in vehicle-treated mice (88 ± 3 mmHg) nor eNOS null mice (104 ± 3 mmHg). In wild-type mice, maximal vasorelaxation of mesenteric arteries to acetylcholine was not altered by chronic l-NAME or induction of diabetes but was reduced by 42 ± 6% in l-NAME-treated diabetic mice. Furthermore, the relative roles of NO and endothelium-derived hyperpolarizing factor (EDHF) in acetylcholine-induced vasorelaxation were altered; the EDHF component was enhanced by l-NAME and blunted by diabetes. These data suggest that NO protects against the development of hypertension during early-stage diabetes in mice, even in the absence of eNOS. Furthermore, in mesenteric arteries, diabetes is associated with reduced EDHF function, with an apparent compensatory increase in NO function. Thus, prior inhibition of NOS results in endothelial dysfunction in early diabetes, since the diabetes-induced reduction in EDHF function cannot be compensated by increases in NO production.

Download Full-text

Phase Separation: From the Initial Nucleation Stage to the Final Ostwald Ripening Regime

MRS Proceedings ◽

10.1557/proc-481-125 ◽

1997 ◽

Vol 481 ◽

Author(s):

Celeste Sagui ◽

Dean Stinson O'Gorman ◽

Martin Grant

Keyword(s):

Phase Separation ◽

Late Stage ◽

Ostwald Ripening ◽

Early Stage ◽

Classical Problem ◽

Nucleation And Growth ◽

New Model ◽

Nucleation Stage ◽

Initial Nucleation

ABSTRACTIn this work we have re-examined the classical problem of nucleation and growth. A new model considers the correlations among droplets and naturally incorporates the crossover from the early-stage, nucleation dominated regime to the scaling, late-stage, coarsening regime within a single framework.

Download Full-text