PERIOD Research Articles

A Distinct Visual Pathway Mediates High-Intensity Light Adaptation of the Circadian Clock in Drosophila.

To provide organisms with a fitness advantage, circadian clocks have to react appropriately to changes in their environment. High-intensity (HI) light plays an essential role in the adaptation to hot summer days, which especially endanger insects of desiccation or prey visibility. Here, we show that solely increasing light intensity leads to an increased midday siesta in Drosophila behavior. Interestingly, this change is independent of the fly's circadian photoreceptor cryptochrome and is solely caused by a small visual organ, the Hofbauer-Buchner eyelets. Using receptor knock-downs, immunostaining, and recently developed calcium tools, we show that the eyelets activate key core clock neurons, namely the s-LNvs, at HI. This activation delays the decrease of PERIOD (PER) in the middle of the day and propagates to downstream target clock neurons that prolong the siesta. We show a new pathway for integrating light-intensity information into the clock network, suggesting new network properties and surprising parallels between Drosophila and the mammalian system.SIGNIFICANCE STATEMENT The ability of animals to adapt to their ever-changing environment plays an important role in their fitness. A key player in this adaptation is the circadian clock. For animals to predict the changes of day and night, they must constantly monitor, detect and incorporate changes in the environment. The appropriate incorporation and reaction to high-intensity (HI) light is of special importance for insects because they might suffer from desiccation during hot summer days. We show here that different photoreceptors have specialized functions to integrate low-intensity, medium-intensity, or HI light into the circadian system in Drosophila These results show surprising parallels to mammalian mechanisms, which also use different photoreceptor subtypes to respond to different light intensities.

Intertextuality in the works of I.P. Shchegolikhin

The article attempts to highlight the problem: can works created in the same historical pe￾riod, based on previous literary experience, form their own style of the era and a special type of image of the author. The authors of the article believe that in the literary text “intertwine”, connecting, signs that go back to different works, elements of other texts-predecessors are combined, as a result of which the text is a “mosaic of quotations “preserving” foreign sounds”, where new lines appear on top of existing ones.In the article, the authors say that the work of I. Shchegolikhin developed in the context of cultural interaction in the cross-border territories of Kazakhstan and Russia, so I. p. Shchegolikhincan rightly be called a representative of Eurasian culture, a Kazakhstani with a special worldview, which is reflected in the implementation of the artistic intent of his works. The author also discusses the issues related to the fact that each work of the writer, building its intertextual field, creates its own history of culture. The au￾thors of this article, relying on the theory of intertextuality and using the elements of intertextual analysis, interpret the individual picture of the world created by the creative imagination of the artist through the prism of the functions of intertextual connections in different works, thereby revealing the peculiarities of the style of a certain era.

CK1α Collaborates with DOUBLETIME to Regulate PERIOD Function in the Drosophila Circadian Clock.

The animal circadian timing system interprets environmental time cues and internal metabolic status to orchestrate circadian rhythms of physiology, allowing animals to perform necessary tasks in a time-of-day-dependent manner. Normal progression of circadian rhythms is dependent on the daily cycling of core transcriptional factors that make up cell-autonomous molecular oscillators. In Drosophila, PERIOD (PER), TIMELESS (TIM), CLOCK (CLK), and CYCLE (CYC) are core clock proteins that function in a transcriptional-translational feedback mechanism to regulate the circadian transcriptome. Posttranslational modifications of core clock proteins provide precise temporal control over when they are active as regulators of clock-controlled genes. In particular, phosphorylation is a key regulatory mechanism that dictates the subcellular localization, stability, and transcriptional activity of clock proteins. Previously, casein kinase 1α (CK1α) has been identified as a kinase that phosphorylates mammalian PER1 and modulates its stability, but the mechanisms by which it modulates PER protein stability is still unclear. Using Drosophila as a model, we show that CK1α has an overall function of speeding up PER metabolism and is required to maintain the 24 h period of circadian rhythms. Our results indicate that CK1α collaborates with the key clock kinase DOUBLETIME (DBT) in both the cytoplasm and the nucleus to regulate the timing of PER-dependent repression of the circadian transcriptome. Specifically, we observe that CK1α promotes PER nuclear localization by antagonizing the activity of DBT to inhibit PER nuclear translocation. Furthermore, CK1α enhances DBT-dependent PER phosphorylation and degradation once PER moves into the nucleus.SIGNIFICANCE STATEMENT Circadian clocks are endogenous timers that integrate environmental signals to impose temporal control over organismal physiology over the 24 h day/night cycle. To maintain the 24 h period length of circadian clocks and to ensure that circadian rhythms are in synchrony with the external environment, key proteins that make up the molecular oscillator are extensively regulated by phosphorylation to ensure that they perform proper time-of-day-specific functions. Casein kinase 1α (CK1α) has previously been identified as a kinase that phosphorylates mammalian PERIOD (PER) proteins to promote their degradation, but the mechanism by which it modulates PER stability is unclear. In this study, we characterize the mechanisms by which CK1α interacts with DOUBLETIME (DBT) to achieve the overall function of speeding up PER metabolism and to ensure proper time-keeping.

The auxin-inducible degradation system enables conditional PERIOD protein depletion in the nervous system of Drosophila melanogaster.

Tools that allow inducible and reversible depletion of target proteins are critical for biological studies. The plant-derived auxin-inducible degradation system (AID) enables the degradation of target proteins tagged with the AID motif. This system has been recently employed in mammalian cells as well as in Caenorhabditis elegans and Drosophila. To test the utility of the AID approach in the nervous system, we used circadian locomotor rhythms as a model and applied the AID method to temporally and spatially degrade PERIOD (PER), a critical pacemaker protein in Drosophila. We found that the period locus can be efficiently tagged with the AID motif by CRISPR/Cas9-based genome editing without disrupting PER function. Moreover, we demonstrated that the AID system could be used to induce rapid and efficient protein degradation in the nervous system as shown by effects on circadian and sleep behaviors. Furthermore, the protein degradation by AID was rapidly reversible after auxin removal. Together, our results show that the AID system provides a powerful tool for behavior studies in Drosophila.

Pro-inflammatory cytokines IL-6 and CCL2 suppress expression of circadian gene Period2 in mammary epithelial cells

Chronic inflammation is known to contribute to tumor initiation and cancer progression. In breast tissue, the core circadian gene Period (PER)2 plays a critical role in mammary gland development and possesses tumor suppressor function. Interleukin (IL)-6 and C-C motif chemokine ligand (CCL) 2 are among the most abundant cytokines in the inflammatory microenvironment. We found that acute stimulation by IL-6/CCL2 reduced PER2 expression in non-tumorigenic breast epithelial cells. Longer term exposure to IL-6/CCL2 suppressed PER2 to an even lower level. IL-6 activated STAT3/NFκB p50 signaling to recruit HDAC1 to the PER2 promoter. CCL2 activated the PI3K/AKT pathway to promote ELK-1 cytoplasm-to-nucleus translocation, recruit HDAC1 to the proximal PER2 promoter and facilitate DNMT3-EZH2-PER2 promoter association. Ectopic expression of PER2 inhibited IL-6 or CCL2 induced mammosphere forming ability and reduced sphere size indicating that PER2 repression in breast epithelial cells can be crucial to activate tumorigenesis in an inflammatory microenvironment. The diminished expression of PER2 can be observed over a time scale of hours to weeks following IL-6/CCL2 stimulation suggesting that PER2 suppression occurs in the early stage of the interaction between an inflammatory microenvironment and normal breast epithelial cells. These data show new mechanisms by which mammary cells interact with a cancerous microenvironment and provide additional evidence that PER2 expression contributes to breast tumorigenesis.

Cryptochrome 2 (CRY2) Suppresses Proliferation and Migration and Regulates Clock Gene Network in Osteosarcoma Cells.

BackgroundCircadian disruption is a potential cancer risk factor in humans. However, the role of the clock gene, cryptochrome 2 (CRY2), in osteosarcoma (OS) is still not clear.Material/MethodsTo evaluate the potential role of CRY2 in HOS osteosarcoma cells, CRY2-silenced cell lines were established. Furthermore, we investigated the effect of CRY2 knockdown on HOS cells by CCK-8, colony formation, migration assay, and flow cytometry, in vitro.ResultsCRY2 knockdown promoted HOS OS cell proliferation and migration. We used a cell cycle assay to show that CRY2 knockdown increased the S phase cell population and reduced the G1 phase cell population. Western blot analyses showed that CRY2 knockdown decreased P53 expression and increased expression of c-myc and cyclin D1. Simultaneously, CRY2 knockdown increased the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2, but did not change the phosphorylation of c-Jun N terminal kinase (JNK) and P38. CRY2 knockdown also increased the expression of matrix metalloproteinase (MMP)-2 and β-catenin, and increased OS cell proliferation and migration by inducing cell cycle progression and promoting mitogen-activated protein kinase (MAPK) and Wnt/β-catenin signaling pathways. Although it has previously been unclear whether the expression of CRY2 affects the expression of other clock genes in the clock gene network, our results show that knockdown of CRY2 significantly increased the mRNA expression of CRY1, Period (PER) 1, PER2, BMAL1, and CLOCK.ConclusionsOur results suggest that CRY2 may be an anti-oncogene in OS, whose functions involve both downstream genes and other circadian genes.

Non-canonical Phototransduction Mediates Synchronization of the Drosophila melanogaster Circadian Clock and Retinal Light Responses

SummaryThe daily light-dark cycles represent a key signal for synchronizing circadian clocks. Both insects and mammals possess dedicated “circadian” photoreceptors but also utilize the visual system for clock resetting. In Drosophila, circadian clock resetting is achieved by the blue-light photoreceptor cryptochrome (CRY), which is expressed within subsets of the brain clock neurons. In addition, rhodopsin-expressing photoreceptor cells contribute to light synchronization. Light resets the molecular clock by CRY-dependent degradation of the clock protein Timeless (TIM), although in specific subsets of key circadian pacemaker neurons, including the small ventral lateral neurons (s-LNvs), TIM and Period (PER) oscillations can be synchronized by light independent of CRY and canonical visual Rhodopsin phototransduction. Here, we show that at least three of the seven Drosophila rhodopsins can utilize an alternative transduction mechanism involving the same α-subunit of the heterotrimeric G protein operating in canonical visual phototransduction (Gq). Surprisingly, in mutants lacking the canonical phospholipase C-β (PLC-β) encoded by the no receptor potential A (norpA) gene, we uncovered a novel transduction pathway using a different PLC-β encoded by the Plc21C gene. This novel pathway is important for behavioral clock resetting to semi-natural light-dark cycles and mediates light-dependent molecular synchronization within the s-LNv clock neurons. The same pathway appears to be responsible for norpA-independent light responses in the compound eye. We show that Rhodopsin 5 (Rh5) and Rh6, present in the R8 subset of retinal photoreceptor cells, drive both the long-term circadian and rapid light responses in the eye.

High-Amplitude Circadian Rhythms in Drosophila Driven by Calcineurin-Mediated Post-translational Control of sarah.

Post-translational control is a crucial mechanism for circadian timekeeping. Evolutionarily conserved kinases and phosphatases have been implicated in circadian phosphorylation and the degradation of clock-relevant proteins, which sustain high-amplitude rhythms with 24-hr periodicity in animal behaviors and physiology. Here, we report a novel clock function of the heterodimeric Ca2+/calmodulin-dependent phosphatase calcineurin and its regulator sarah (sra) in Drosophila Genomic deletion of the sra locus dampened circadian locomotor activity rhythms in free-running constant dark after entrainment in light-dark cycles. Poor rhythms in sra mutant behaviors were accompanied by lower expression of two oscillating clock proteins, PERIOD (PER) and TIMELESS (TIM), at the post-transcriptional level. RNA interference-mediated sra depletion in circadian pacemaker neurons was sufficient to phenocopy loss-of-function mutation in sra On the other hand, a constitutively active form of the catalytic calcineurin subunit, Pp2B-14DACT, shortened circadian periodicity in locomotor behaviors and phase-advanced PER and TIM rhythms when overexpressed in clock neurons. Heterozygous sra deletion induced behavioral arrhythmicity in Pp2B-14DACT flies, whereas sra overexpression rescued short periods in these animals. Finally, pharmacological inhibition of calcineurin in either wild-type flies or clock-less S2 cells decreased the levels of PER and TIM, likely by facilitating their proteasomal degradation. Taken together, these data suggest that sra negatively regulates calcineurin by cell-autonomously titrating calcineurin-dependent stabilization of PER and TIM proteins, thereby sustaining high-amplitude behavioral rhythms in Drosophila.

Psychometric Properties of the Sexual Event Diary in a Sample of Dutch Women With Female Sexual Interest/Arousal Disorder

BackgroundEfficacy of on-demand drugs for women with hypoactive sexual desire disorder or female sexual interest/arousal disorder (FSIAD) should be assessed using a validated instrument that assesses the discrete sexual events during which the on-demand drug is taken, because this type of assessment is more proximate to an on-demand drug's efficacy compared to instruments that assess sexual function over longer periods of time. AimThe aim of this study was to assess the psychometric properties of the Dutch translation of the previously validated 11-item Sexual Event Diary (SED) for measuring sexual satisfaction and sexual functioning during discrete sexual events. MethodsPsychometric assessment was performed on data of 1,840 SEDs from 139 women with hypoactive sexual desire disorder/FSIAD, collected during a randomized clinical cross-over trial conducted in the Netherlands. OutcomesItem scores of the SED at the event level, and at subject level, summarized item scores during the placebo run-in period (PRI) and active treatment period, and score changes from PRI to active treatment period. ResultsReliability and convergent validity were confirmed. All item scores showed the ability to discriminate between known groups. Larger mean score changes from PRI were observed in groups with known benefit from the medication, as compared to those with no benefit. Guyatt effect sizes ranged from 0.51–1.02, thereby demonstrating ability to detect change. Clinical TranslationThe Dutch version of the SED is an excellent instrument for assessing female sexual functioning and sexual satisfaction during discrete sexual events and for assessing these concepts over longer periods of time. ConclusionsData were collected in a randomized, well-controlled trial. The large number of data points gave high statistical power, and the results confirmed previous findings. However, care is needed when generalizing the SED's validity to other areas of research, eg, recreational drug use and sexual risky behaviors, since the current validation study has not used such data. Consistent with the US-English version, the Dutch version of the SED is a reliable, valid, and responsive instrument, and suitable for use in evaluating effects of on-demand drugs in women with FSIAD.van Nes Y, Bloemers J, Kessels R, et al. Psychometric Properties of the Sexual Event Diary in a Sample of Dutch Women With Female Sexual Interest/Arousal Disorder. J Sex Med 2018;15:722–731.

CK1/Doubletime activity delays transcription activation in the circadian clock.

In the Drosophila circadian clock, Period (PER) and Timeless (TIM) proteins inhibit Clock-mediated transcription of per and tim genes until PER is degraded by Doubletime/CK1 (DBT)-mediated phosphorylation, establishing a negative feedback loop. Multiple regulatory delays within this feedback loop ensure ~24 hr periodicity. Of these delays, the mechanisms that regulate delayed PER degradation (and Clock reactivation) remain unclear. Here we show that phosphorylation of certain DBT target sites within a central region of PER affect PER inhibition of Clock and the stability of the PER/TIM complex. Our results indicate that phosphorylation of PER residue S589 stabilizes and activates PER inhibitory function in the presence of TIM, but promotes PER degradation in its absence. The role of DBT in regulating PER activity, stabilization and degradation ensures that these events are chronologically and biochemically linked, and contributes to the timing of an essential delay that influences the period of the circadian clock.

Phosphorylation of SLIMB by Minibrain/DYRK1a Activates SLIMB‐mediated Circadian Clock Protein Degradation

SLIMB/β‐TrCP is a F‐box protein that provides substrate specificity to E3 ubiquitin ligase protein complexes, which ubiquitinate cellular proteins and target them for degradation by the 26S proteasome. Timely degradation of substrate proteins is intricately linked and critical to the regulation of numerous molecular processes, including cell cycle regulation, developmental timing, tumorigenesis, metabolism, and circadian biology. Previous work in our laboratory characterized the requirements for the interaction between SLIMB and PERIOD (PER), a key transcription factor that regulates circadian gene expression. The level of PER oscillates in a circadian manner and loss of SLIMB function disrupts PER cycling, leading to misregulation of the circadian oscillator. In contrast to the numerous studies examining the post‐translation modification (PTM) of substrate proteins to enable recognition by SLIMB, e.g. interaction of SLIMB and PER, there is a deficit of literature that investigates the PTM of F‐box proteins that modulate their function. To understand the post‐translational regulation of SLIMB function, we have identified 7 phosphorylation sites on SLIMB using mass spectrometry proteomics and characterized the function of these SLIMB PTMs in the context of the Drosophila circadian clock system.Support or Funding InformationNIH R01 GM102225 to JCC, F31 DK104624 to AJC, and T32 GM007377This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Kidney‐Specific KO of the Circadian Clock Protein BMAL1 Lowers Blood Pressure in Male C57BL/6J Mice

All living organisms have an internal circadian clock that regulates the timing of many physiological functions. Dysregulation of circadian rhythms is associated with cardiovascular disease and cancer. Blood pressure (BP) has a circadian rhythm with a peak during the active phase and a dip during the rest phase. The core of the circadian clock consists of four core transcription factors: BMAL1, CLOCK, CRYTOCHROME (CRY), and PERIOD (PER). BMAL1 and CLOCK bind as a heterodimer to E‐box response elements of 40% of all expressed genes to regulate expression. PER and CRY act in a negative feedback loop, inhibiting the actions of BMAL1 and CLOCK. In 2007, Curtis et al. reported that global Bmal1 KO male mice had a 10 mmHg decrease in their mean arterial pressure (MAP) and lost their circadian rhythm of BP relative to control mice. Evidence from our laboratory and others suggest there is a link between the circadian clock in the kidney and BP regulation. We hypothesized that BMAL1 is critical for proper regulation of renal gene expression and that kidney‐specific KO of Bmal1 would alter BP. We generated kidney‐specific Bmal1 KO mice using Ksp‐cadherin Cre, which typically results in a distal nephron‐specific KO. Continuous BP measurements were made in the Cre+ kidney‐specific Bmal1 KO mice (KS‐Bmal1 KO) and the Cre‐ littermate control mice (CNTL) using radiotelemetry. Female CNTL and KS‐Bmal1 KO mice did not exhibit a detectable difference in MAP (~106 mm Hg in both genotypes, p=0.93, n=6 mice per group). Male CNTL mice exhibited a 24 hr MAP of 110 mm Hg whereas male KS‐Bmal1 KO mice had a 24 hr MAP of 99 mm Hg (P<0.001, n=5 CNTL, 7 KO). Kidney‐specific KO of Bmal1 did not appear to alter the circadian rhythm of BP in either gender. In response to a high salt diet plus mineralocorticoid treatment (HS/DOCP), male KS‐Bmal1 KO mice MAP increased by 10 mm Hg whereas the MAP for CNTL mice only increased by 4 mm Hg (P<0.01, n=3 CNTL, 7 KO). Female KS‐Bmal1 KO mice MAP did not change in response to HS/DOCP. These data suggest that kidney‐specific KO of Bmal1 increases sensitivity of male mice to salt and mineralocorticoid treatment. Preliminary metabolic cage data demonstrated that male KS‐Bmal1 KO mice retained urinary sodium during the first two days of HS/DOCP treatment whereas CNTL male mice continued to excrete increasing amounts of sodium (P<0.05, n=3 mice per genotype). The present study demonstrated that BMAL1 in the kidney likely acts in a sex‐specific manner and may not be important for circadian fluctuations in BP under the conditions tested. Together these results suggest that BMAL1 plays a critical role in the regulation of renal sodium reabsorption with important consequences for overall BP control.Support or Funding InformationNIH NIDDK R01DK109570 awarded to Michelle Gumz; NIH T32DK104721 awarded to Lauren Douma; American Heart Association Grant‐in‐aid awarded to Michelle Gumz; Gatorade Trust, Division of Nephrology, Department of Medicine, University of FloridaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

On Variations in the Level of PER in Glial Clocks of Drosophila Optic Lobe and Its Negative Regulation by PDF Signaling.

We show that the level of the core protein of the circadian clock Period (PER) expressed by glial peripheral oscillators depends on their location in the Drosophila optic lobe. It appears to be controlled by the ventral lateral neurons (LNvs) that release the circadian neurotransmitter Pigment Dispersing Factor (PDF). We demonstrate that glial cells of the distal medulla neuropil (dMnGl) that lie in the vicinity of the PDF-releasing terminals of the LNvs possess receptors for PDF (PDFRs) and express PER at significantly higher level than other types of glia. Surprisingly, the amplitude of PER molecular oscillations in dMnGl is increased twofold in PDF-free environment, that is in Pdf0 mutants. The Pdf0 mutants also reveal an increased level of glia-specific protein REPO in dMnGl. The photoreceptors of the compound eye (R-cells) of the PDF-null flies, on the other hand, exhibit de-synchrony of PER molecular oscillations, which manifests itself as increased variability of PER-specific immunofluorescence among the R-cells. Moreover, the daily pattern of expression of the presynaptic protein Bruchpilot (BRP) in the lamina terminals of the R-cells is changed in Pdf0 mutant. Considering that PDFRs are also expressed by the marginal glia of the lamina that surround the R-cell terminals, the LNv pacemakers appear to be the likely modulators of molecular cycling in the peripheral clocks of both the glial cells and the photoreceptors of the compound eye. Consequently, some form of PDF-based coupling of the glial clocks and the photoreceptors of the eye with the central LNv pacemakers must be operational.

Infection by Xanthomonas campestris pv.viticola under temperature increase and carbon dioxide concentrations

The experiments were carried out under controlled conditions to evaluate the impact of increased temperature and concentration of carbon dioxide on infection of Xanthomonas campestris pv viticola, the causal agent of bacterial canker in Vine seedlings. It proceeded the evaluation of the following epidemiological components: incubation period (PI), severity (SEV) and using that data were calculated the area under the disease progress curve (AUDPC). It used grape seedlings (Italia, Crimson Seedless, Sugraone and Selection 8) inoculated with bacterial suspension (108 CFU mL-1). The experimental design was completely randomized, factorial arrangement 4x4 (cultivar x temperature) and 4x2 (cultivar x carbon dioxide concentration) which was carried out twice. The data were subjected to variance analysis. Increasing temperature reduced bacterium’s incubation period with significant differences between genotypes. For Selection 8 and Crimson temperature increase caused enhancement on severity and AUDPC. For Seleção 8 the incubation period (PI) was extended from 7.93 to 30.18 days when the concentration changed from 390 to 770 µmol/mol. The increased CO2 concentration reduced AACPSD and SEV for Sugraone and Selection 8. The results show that the temperature and carbon dioxide (CO2) concentration of the air may have different effects on bacterial canker of grapevine.

Shedding Light on Night Owl Behavior: How the Disordered C-Terminal Tail of CRY1 Modulates Circadian Timekeeping

Circadian rhythms are driven by a negative feedback loop in which the heterodimer CLOCK:BMAL1 promotes transcription of clock-controlled genes, including its own repressors Period (PER) and Cryptochrome (CRY). The deregulation of circadian rhythms can lead to delayed sleep phase disorder (DSPD), where the afflicted “night owls” are only able to fall asleep hours after midnight and have difficulty waking up early in the morning. CRY proteins contain a photolyase homology region (PHR) and an intrinsically disordered C-terminal tail; previous studies identified an essential role for the CRY1 PHR in generating circadian rhythms, while deletion of the tail was shown to alter the length of the circadian period. Our lab recently identified how the CRY1 PHR binds to both CLOCK and BMAL1 to induce repression and close the negative feedback loop. To date, little is known about how the CRY1 PHR and tail might interact to control this process. One prevalent mutation in DSPD patients leads to deletion of CRY1 Exon 11, a region that encodes for 25 amino acids in the tail, resulting in enhanced co-immunoprecipitation by CLOCK:BMAL1 and a longer circadian period. We hypothesized that the CRY1 tail might bind directly to the PHR to regulate interactions with CLOCK or BMAL1. Here we show that the C-terminal tail and CLOCK compete for binding to the CRY1 PHR, and that the peptide encoded by Exon 11 is sufficient for this interaction. Therefore, our data demonstrate how the tail can directly regulate CRY1 interactions with CLOCK:BMAL1 to provide initial insight into the molecular basis of DSPD.

Use of Solar Energy Hybrid Dryer with Techno-Ergonomic Application to Increase Productivity of Dodol Wokers in Buleleng, Bali

Penglatan Village is one of dodol industrial centres in Buleleng Regency, Bali. Dodol is a balinese traditional snack that usually used for religion ceremonies offering or “sesajen”. Dodol’s making processes had several layer of stages, from making dough, stirring dough, packaging and drying dodol. Because the drying process is done with traditional work tools, this process cause several ergonomic problems. Based on preliminary research, the complaints that felt by dodol workers are pain in the neck, shoulders, back, waist, head, and hands. Therefore, productivity of workers are decreasing. This problem solved by designing a hybrid solar drying tools with the application of techno-ergonomic, appropriate technology concept (TTG) and from ergonomic science with the application of SHIP concept (systemic, holistic, interdisciplinary, participatory). This research use 20 people as sample. The sample performance is observed while working traditionally and using techno-ergonomic hybrid solar dryer tools. The measurements conducted three times which are First Period (PI), Second Period (PII), and Third Period (PIII) with interspersed time for rest. WOP (washing out period) is intended to eliminate residual effects. The data were analyzed with SPSS program with significance level of 0.05. Hopefully, with this solution will improve worker productivity.

Circadian clock and oral cancer.

The circadian clock is comprised of a master component situated in the hypothalamic suprachiasmatic nucleus and subordinate clock genes in almost every cell of the body. The circadian clock genes and their encoded proteins govern the organism to follow the natural signals of time, and adapt to external changes in the environment. The majority of physiological processes in mammals exhibit variable circadian rhythms, which are generated and coordinated by an oscillation in the expression of the clock genes. A number of studies have reported that alteration in the expression level of clock genes is correlated with several pathological conditions, including cancer. However, little is known about the role of clock genes in homeostasis of the oral epithelium and their disturbances in oral carcinogenesis. The present review summarizes the current state of knowledge of the implications of clock genes in oral cancer. It has been demonstrated that the development of oral squamous cell carcinoma undergoes circadian oscillation in relation to tumor volume and proliferation rate. The circadian clock gene period (PER)1 has been associated with oral cancer pathogenesis and it is suggested that changes in the expression of PER1 may exhibit an important role in the development, invasion, and metastasis of oral squamous cell carcinoma. However, its role remains elusive and there is a need for further research in order to understand the underlying mechanisms of the clock genes in oral cancer pathogenesis.

Evaluating the Autonomy of the Drosophila Circadian Clock in Dissociated Neuronal Culture.

Circadian behavioral rhythms offer an excellent model to study intricate interactions between the molecular and neuronal mechanisms of behavior. In mammals, pacemaker neurons in the suprachiasmatic nucleus (SCN) generate rhythms cell-autonomously, which are synchronized by the network interactions within the circadian circuit to drive behavioral rhythms. However, whether this principle is universal to circadian systems in animals remains unanswered. Here, we examined the autonomy of the Drosophila circadian clock by monitoring transcriptional and post-transcriptional rhythms of individual clock neurons in dispersed culture with time-lapse microscopy. Expression patterns of the transcriptional reporter show that CLOCK/CYCLE (CLK/CYC)-mediated transcription is constantly active in dissociated clock neurons. In contrast, the expression profile of the post-transcriptional reporter indicates that PERIOD (PER) protein levels fluctuate and ~10% of cells display rhythms in PER levels with periods in the circadian range. Nevertheless, PER and TIM are enriched in the cytoplasm and no periodic PER nuclear accumulation was observed. These results suggest that repression of CLK/CYC-mediated transcription by nuclear PER is impaired, and thus the negative feedback loop of the molecular clock is incomplete in isolated clock neurons. We further demonstrate that, by pharmacological assays using the non-amidated form of neuropeptide pigment-dispersing factor (PDF), which could be specifically secreted from larval LNvs and adult s-LNvs, downstream events of the PDF signaling are partly impaired in dissociated larval clock neurons. Although non-amidated PDF is likely to be less active than the amidated one, these results point out the possibility that alteration in PDF downstream signaling may play a role in dampening of molecular rhythms in isolated clock neurons. Taken together, our results suggest that Drosophila clocks are weak oscillators that need to be in the intact circadian circuit to generate robust 24-h rhythms.

SIK3–HDAC4 signaling regulates Drosophila circadian male sex drive rhythm via modulating the DN1 clock neurons

The physiology and behavior of many organisms are subject to daily cycles. In Drosophila melanogaster the daily locomotion patterns of single flies are characterized by bursts of activity at dawn and dusk. Two distinct clusters of clock neurons-morning oscillators (M cells) and evening oscillators (E cells)-are largely responsible for these activity bursts. In contrast, male-female pairs of flies follow a distinct pattern, most notably characterized by an activity trough at dusk followed by a high level of male courtship during the night. This male sex drive rhythm (MSDR) is mediated by the M cells along with DN1 neurons, a cluster of clock neurons located in the dorsal posterior region of the brain. Here we report that males lacking Salt-inducible kinase 3 (SIK3) expression in M cells exhibit a short period of MSDR but a long period of single-fly locomotor rhythm (SLR). Moreover, lack of Sik3 in M cells decreases the amplitude of PERIOD (PER) cycling in DN1 neurons, suggesting that SIK3 non-cell-autonomously regulates DN1 neurons' molecular clock. We also show that Sik3 reduction interferes with circadian nucleocytoplasmic shuttling of Histone deacetylase 4 (HDAC4), a SIK3 phosphorylation target, in clock neurons and that constitutive HDAC4 localization in the nucleus shortens the period of MSDR. Taking these findings together, we conclude that SIK3-HDAC4 signaling in M cells regulates MSDR by regulating the molecular oscillation in DN1 neurons.

ІМУНОЛОГІЧНИЙ МОНІТОРИНГ ДОРОСЛОГО НАСЕЛЕННЯ ЛЬВІВЩИНИ ЩОДО КАШЛЮКУ У 2016 РОЦІ

Мета – провести серологічний моніторинг серед дорослого населення щодо кількісного визначення імуноглобулінів класу G до кашлюку.Матеріали і методи. Дослідження проводились з використанням методу імуноферментного аналізу.Результати досліджень та їх обговорення. Серонегативний прошарок населення становить 50,0 %. Специфічні імуноглобуліни G у високих титрах (понад 50 BU/ml) були виявлені у 3,75 % осіб, що свідчить про період реконвалесценції після кашлюку.Висновки. Використання цього методу підтверджує наявність випадків захворювання на кашлюк серед дорослого населення при недостатній настороженості медичних працівників щодо захворюваності на кашлюк даної вікової групи. Підтверджено об’єктивність і інформативність методу та доцільність його широкого впровадження в практику громадського здоров’я з метою здійснення належного епідеміологічного нагляду.