NF-κB modifies the mammalian circadian clock through interaction with the core clock protein BMAL1

In mammals, the circadian clock coordinates cell physiological processes including inflammation. Recent studies suggested a crosstalk between these two pathways. However, the mechanism of how inflammation affects the clock is not well understood. Here, we investigated the role of the proinflammatory transcription factor NF-κB in regulating clock function. Using a combination of genetic and pharmacological approaches, we show that perturbation of the canonical NF-κB subunit RELA in the human U2OS cellular model altered core clock gene expression. While RELA activation shortened period length and dampened amplitude, its inhibition lengthened period length and caused amplitude phenotypes. NF-κB perturbation also altered circadian rhythms in the master suprachiasmatic nucleus (SCN) clock and locomotor activity behavior under different light/dark conditions. We show that RELA, like the clock repressor CRY1, repressed the transcriptional activity of BMAL1/CLOCK at the circadian E-box cis-element. Biochemical and biophysical analysis showed that RELA binds to the transactivation domain of BMAL1. These data support a model in which NF-kB competes with CRY1 and coactivator CBP/p300 for BMAL1 binding to affect circadian transcription. This is further supported by chromatin immunoprecipitation analysis showing that binding of RELA, BMAL1 and CLOCK converges on the E-boxes of clock genes. Taken together, these data support a significant role for NF-κB in directly regulating the circadian clock and highlight mutual regulation between the circadian and inflammatory pathways.

Does the Length of the Cooling-off Period Affect Audit Quality?

This paper examines the association between the length of the cooling-off period and audit quality: (1) when partners rotate back and (2) during the cooling-off period, ahead of an extension to the minimum cooling-off period requirement in Australia. Using multiple measures of audit quality, we find some evidence of a positive association between the cooling-off period length and audit quality when partners rotate back, yet evidence of a negative association between the two, during the cooling-off period. We also find that auditor and client characteristics-such as partner busyness, client knowledge, geographic proximity, and client importance-play important roles in determining the cooling-off period length and whether a partner rotates back onto a client. Overall, we provide timely evidence that extending the cooling-off period only marginally enhances audit quality when a partner rotates back onto a client, and evidence of an unintended consequence of this policy during the cooling-off period.

EP.WE.782Retrospective analysis of acute surgical admissions during COVID-19 pandemic

Background COVID-19 pandemic has affected the number of surgical admissions and the number of emergency general surgical operations performed. COVID-19 pandemic has also led to changes in how some of the acute surgical patients were managed. Aim of the study was to compare acute surgical admissions and number of emergency general surgical procedures in this period. Material and Method We retrospectively analyzed acute surgical admissions during the pandemic from 20/3/20 to 19/4/20 and compared it with acute surgical admissions during pre-COVID-19 period from 1/11/2019 to 30/11/2019. Results During the COVID-19 pandemic 97 patients were assessed and admitted by the General surgical team, during the pre COVID-19 period 205 patients were assessed and admitted by the General surgical team. The number of acute surgical admissions during COVID-19 pandemic dropped by 53%. There were 46% less emergency surgeries performed during COVID 19 pandemic period. Length of stay during and before the COVID-19pandemic were 4.1 vs 4.4 days. Conclusion During the COVID-19 pandemic number of acute surgical admissions and the number of emergency surgeries were fewer than during pre COVID-19 pandemic. Length of hospital stay was less during COVID-19 pandemic.

Pengaruh lama kering kandang dan periode laktasi terhadap produksi dan kualitas susu awal laktasi pada sapi perah

<p class="MDPI17abstract"><strong>Objective: </strong>This study aims to determine the relationship between length of dry - and lactation period on milk production of early lactation, as well as milk fat, lactose, protein and specific gravity.</p><p class="MDPI17abstract"><strong>Methods: </strong>A total of 24 dry period Friesian Holstein cows at the 7th month of pregnancy were used. The dry period length were divided into 4 groups and consists of 3 periods of lactation. Parameters determined were milk production and quality in the first month of lactation. Data were tested using the Kruskal Wallis for the effect of dry period length and lactation period on milk production and quality. Wilcoxon analysis for continued trials of significant results.<strong></strong></p><p class="MDPI17abstract"><strong>Results</strong><strong>: </strong>The results showed that 40-79 days dry period length had no significant effect on milk production (P= 0.854), milk specific gravity (P= 0.922), milk fat (P= 0.339), milk protein (P= 0.392) and milk lactose (P= 0.635). Lactation period has a significant effect on milk production (P= 0.001), milk fat (P= 0.011), milk protein (P= 0.028), milk lactose (P= 0.004), but has no significant effect on milk density (P= 0.683). The highest milk production was produced in the VI period with milk production at 675.17 liters/ month (P= 0.001).<strong></strong></p><strong>Conclusions: </strong>The dry period of 40 to 79 days gives similar milk production and quality. The lactation period affects the amount and quality of milk at early lactation. Lactating cows reared in good management are able to produce milk with good quality and high production until the-VI of lactation period.

Chloride oscillation in pacemaker neurons regulates circadian rhythms through a chloride-sensing WNK kinase signaling cascade

Central pacemaker neurons regulate circadian rhythms and undergo diurnal variation in electrical activity in mammals and flies. In mammals, circadian variation in the intracellular chloride concentration of pacemaker neurons has been proposed to influence the response to GABAergic neurotransmission through GABAA receptor chloride channels. However, results have been contradictory, and a recent study demonstrated circadian variation in pacemaker neuron chloride without an effect on GABA response. Therefore, whether and how intracellular chloride regulates circadian rhythms remains controversial. Here, we demonstrate a signaling role for intracellular chloride in the Drosophila ventral lateral (LNv) pacemaker neurons. In control flies, intracellular chloride increases in LNv neurons over the course of the morning. Chloride transport through the sodium-potassium-2-chloride (NKCC) and potassium-chloride (KCC) cotransporters is a major determinant of intracellular chloride concentrations. Drosophila melanogaster with loss-of-function mutations in the NKCC encoded by Ncc69 have abnormally low intracellular chloride six hours after lights on, and a lengthened circadian period. Loss of kcc, which is expected to increase intracellular chloride, suppresses the long-period phenotype of Ncc69 mutant flies. Activation of a chloride-inhibited kinase cascade, consisting of the WNK (With No Lysine (K)) kinase and its downstream substrate, Fray, is necessary and sufficient to prolong period length. Fray activation of an inwardly rectifying potassium channel, Irk1, is also required for the long-period phenotype. These results indicate that the NKCC-dependent rise in intracellular chloride in Drosophila LNv pacemaker neurons restrains WNK-Fray signaling and overactivation of an inwardly rectifying potassium channel to maintain normal circadian period length.

FRQ-CK1 Interaction Underlies Temperature Compensation of the Neurospora Circadian Clock

Temperature compensation allows clocks to adapt to all seasons by having a relatively constant period length at different physiological temperatures, but the mechanism of temperature compensation is unclear. Stability of clock proteins was previously proposed to be a major factor that regulated temperature compensation.