Development of Circadian Oscillators in Neurosphere Cultures during Adult Neurogenesis

Circadian rhythms are common in many cell types but are reported to be lacking in embryonic stem cells. Recent studies have described possible interactions between the molecular mechanism of circadian clocks and the signaling pathways that regulate stem cell differentiation. Circadian rhythms have not been examined well in neural stem cells and progenitor cells that produce new neurons and glial cells during adult neurogenesis. To evaluate circadian timing abilities of cells undergoing neural differentiation, neurospheres were prepared from the mouse subventricular zone (SVZ), a rich source of adult neural stem cells. Circadian rhythms in mPer1 gene expression were recorded in individual spheres, and cell types were characterized by confocal immunofluorescence microscopy at early and late developmental stages in vitro. Circadian rhythms were observed in neurospheres induced to differentiate into neurons or glia, and rhythms emerged within 3–4 days as differentiation proceeded, suggesting that the neural stem cell state suppresses the functioning of the circadian clock. Evidence was also provided that neural stem progenitor cells derived from the SVZ of adult mice are self-sufficient clock cells capable of producing a circadian rhythm without input from known circadian pacemakers of the organism. Expression of mPer1 occurred in high frequency oscillations before circadian rhythms were detected, which may represent a role for this circadian clock gene in the fast cycling of gene expression responsible for early cell differentiation

Development of Circadian Oscillators in Neurosphere Cultures during Adult Neurogenesis

Circadian rhythms are common in many cell types but are reported to be lacking in embryonic stem cells. Recent studies have described possible interactions between the molecular mechanism of circadian clocks and the signaling pathways that regulate stem cell differentiation. Circadian rhythms have not been examined well in neural stem cells and progenitor cells that produce new neurons and glial cells during adult neurogenesis. To evaluate circadian timing abilities of cells undergoing neural differentiation, neurospheres were prepared from the mouse subventricular zone (SVZ), a rich source of adult neural stem cells. Circadian rhythms in mPer1 gene expression were recorded in individual spheres, and cell types were characterized by confocal immunofluorescence microscopy at early and late developmental stages in vitro. Circadian rhythms were observed in neurospheres induced to differentiate into neurons or glia, and rhythms emerged within 3–4 days as differentiation proceeded, suggesting that the neural stem cell state suppresses the functioning of the circadian clock. Evidence was also provided that neural stem progenitor cells derived from the SVZ of adult mice are self-sufficient clock cells capable of producing a circadian rhythm without input from known circadian pacemakers of the organism. Expression of mPer1 occurred in high frequency oscillations before circadian rhythms were detected, which may represent a role for this circadian clock gene in the fast cycling of gene expression responsible for early cell differentiation

WASTE MANAGEMENT IN THE KAWATUNA LANDFILL SITE OF PALU CITY

Abstract: The waste is all kind of things or material/human excreta, animal, vegetation or anything from the result of human activity to fulfill their daily need. This waste may trigger and or cause contamination to the water, land, and air and cause damage to the human environmental. The ultimate waste disposal of the city in The Landfill Site is against some obstacles, physically and nonphysical, such as social, economy, maintenance problems, etc. According the field experience in some area especially in the City of Palu, city waste management in TPA Kawatuna or Kawatuna Landfill Site consistently practice open dumping system with specific awareness on the environmental protection. The problem occur in Kawatuna Landfill Site is none of waste selection criteria. This condition caused a habitant of the scavengers to earn the life for their expanse, but this settlement has negative impact to their healthiness. The contours of landfill site are valley and hilly. This caused a waste collections activity from the waste employee throw away randomly to the valley of landfill site area and caused wider landfill area. In addition, another wider valley becomes the garbage collection area and causes destruction to the land structure. As a result, the qualitative descriptive method of this research concludes applicable waste management system for the Kawatuna Landfill Site and factors that influences the waste management system. Effort and good cooperation are necessary for a good waste management practice in Kawatuna Landfill Site. This practice starts from each of us where the waste is a requirement that should minimize together. Application of 4R (Reduce, Replace, Reuse and Recycle) is the first step in maximizing the waste management system of Kawatuna Landfill Site. All of this effort certainly requires a support from human resources, facility and infrastructure, social participation and government regulation. Keywords: Waste, Waste Management, Kawatuna Landfill Abstrak: Sampah ialah semua jenis benda atau barang bangunan/kotoran manusia, hewan atau tumbuh-tumbuhan atau yang berasal dari aktivitas kehidupan manusia dalam memenuhi kebutuhan hidupnya yang dapat menimbulkan dan atau mengakibatkan pengotoran terhadap air, tanah dan udara sehingga dapat menimbulkan pengrusakan lingkungan hidup manusia. Penampungan akhir sampah kota dilakukan di Tempat Pembuangan Akhir (TPA) mengalami berbagai macam kendala baik fisik maupun non fisik, seperti masalah sosial, ekonomi, pemeliharaan dan lain–lain. Dari berbagai kenyataan yang ada di lapangan, di berbagai daerah, khususnya di Kota Palu, pengelolaan sampah perkotaan di TPA Kawatuna masih menggunakan sistem pembuangan terbuka (open dumping), dimana sistem ini kurang memperhatikan aspek perlindungan lingkungan. Masalah yang timbul di TPA Kawatuna yakni : belum ada pengelolaan untuk pemilahan jenis sampah, kondisi ini mengakibatkan munculnya permukiman para pemulung yang mencari nafkah dengan memilah sampah yang dapat memberikan pendapatan bagi mereka, namun keberadaan permukiman mereka dapat memberikan pengaruh negatif bagi kesehatan mereka. Dan kondisi TPA yang berkontur yaitu lembah dan perbukitan menjadikan sampah yang telah dikumpulkan oleh petugas dibuang sembarangan pada lembah di daerah TPA yang mengakibatkan bertambah luasnya daratan sampah, hal lain lembah yang cukup besar dijadikan sebagai penampung sampah yang dapat menyebabkan rusaknya struktur tanah. Olehnya penyelesaian penelitian dengan menggunakan metode deskriptif kualitatif didapatkan sistem pengelolaan sampah yang sesuai diterapkan di TPA Kawatuna dan faktor – faktor yang mempengaruhi sistem pengelolaan sampah. Diperlukan usaha dan kerjasama yang baik agar pengolahan sampah di TPA Kawatuna dapat berjalan dengan baik. Hal tersebut dimulai dari diri kita masing – masing bahwa sampah merupakan suatu kebutuhan yang harus kita minimalisasikan bersama – sama. Penerapan 4R yakni (Reduce, Replace, Reuse and Recycle) merupakan langkah awal agar sistem pengelolaan sampah di TPA Kawatuna berjalan maksimal. Tentunya semua itu perlu didukung dengan sumber daya manusia, sarana prasarana, partisipasi masyarakat dan peraturan pemerintah. Kata Kunci: Sampah, Pengelolaan Sampah, TPA Kawatun

Allied health and complementary therapy usage in Australian women with chronic pelvic pain : a cross-sectional study

Background: Chronic pelvic pain (CPP) causes non-cyclical pelvic pain, period pain, fatigue and other painful symptoms. Current medical and surgical management strategies are often not sufficient to manage these symptoms and may lead to uptake of other therapies. Aims: To determine the prevalence of allied health (AH) and complementary therapy (CM) use, the cost burden of these therapies and explore predictive factors for using allied health or complementary medicines. Materials and methods: An online cross-sectional questionnaire using the WERF EndoCost tool was undertaken between February to April 2017. People were eligible to participate in the survey if they were aged 18–45, living in Australia and had chronic pelvic pain. Results: From 409 responses, 340/409 (83%) of respondents reported a diagnosis of endometriosis. One hundred and five (30%) women with self-reported endometriosis, and thirteen (18%) women with other forms of CPP saw at least one AH or CM practitioner in the previous two months, with physiotherapists and acupuncturists the most common. Women who accessed CM or AH services spent an average of $480.32 AUD in the previous two months. A positive correlation was found between education and number of AH or CM therapies accessed in the past two months (p<0.001) and between income level and number of therapists (p=0.028). Conclusions: Women with CPP commonly access AH and CM therapies, with a high out of pocket cost. The high cost and associations with income and education levels may warrant a change to policy to improve equitable access to these services

Circadian Clocks in Neural Stem Cells and their Modulation of Adult Neurogenesis, Fate Commitment, and Cell Death

Adult neurogenesis creates new neurons and glial cells from stem cells in the human brain throughout life, and it is best understood in the dentate gyrus (DG) of the hippocampus and the subventricular zone (SVZ). Recent studies have described possible interactions between the molecular mechanism of circadian clocks and the signaling pathways that regulate stem cell differentiation. Circadian rhythms have been identified in the olfactory bulb and the hippocampus, but the role of any endogenous circadian oscillator cells in neurogenesis and their importance in learning and memory remains unclear. Circadian rhythms have not been examined well in neural stem cells and progenitor cells that produce new neurons and glial cells during adult neurogenesis. To evaluate circadian timing abilities of cells undergoing neural differentiation, neurospheres were prepared from mPer1::luc mouse SVZ and DG. Circadian bioluminescence rhythms were recorded in neurospheres maintained in culture medium that induces neurogenesis but not in one that maintains the stem cell state. Cell types were also characterized by confocal immunofluorescence microscopy at early and late developmental stages in vitro. Evidence was provided that neural stem progenitor cells (NSPCs) derived from the SVZ and DG of adult mice are self-sufficient clock cells capable of producing a circadian rhythm without input from known circadian pacemakers of the organism. Extremely rare percentages of mature neuronal cells were observed during ontogeny of rhythms. The bulk of the neurosphere cells were undifferentiated, indicating that they are the circadian clock cells producing timing signals. This conclusion was supported by immunocytochemistry for mPER1 protein that was localized to the inner, more stem cell-like neurosphere core. To further test whether circadian clocks in NSPCs are necessary for growth, differentiation and cell survival, neurospheres were cultured from Bmal1-/- and Cry1-/-,2-/- knockout mice. Neurosheres from Bmal1-/- knockout mice displayed unusually high differentiation into glia rather than neurons according to GFAP and NeuN expression, respectively, and very few DCX and BetaIII tubulin-positive, immature neurons were observed. The knockout neurospheres also had areas visibly devoid of cells and overall higher cell death. Neurospheres from mice lacking Cry1 and Cry2 showed significantly reduced growth. Altered NSPC proliferation and differentiation in these mice may impede memory formation and could provide a way to identify circadian timing effects in neurodegenerative disorders and impaired brain functions

Circadian Clocks in Neural Stem Cells and their Modulation of Adult Neurogenesis, Fate Commitment, and Cell Death

Adult neurogenesis creates new neurons and glial cells from stem cells in the human brain throughout life, and it is best understood in the dentate gyrus (DG) of the hippocampus and the subventricular zone (SVZ). Recent studies have described possible interactions between the molecular mechanism of circadian clocks and the signaling pathways that regulate stem cell differentiation. Circadian rhythms have been identified in the olfactory bulb and the hippocampus, but the role of any endogenous circadian oscillator cells in neurogenesis and their importance in learning and memory remains unclear. Circadian rhythms have not been examined well in neural stem cells and progenitor cells that produce new neurons and glial cells during adult neurogenesis. To evaluate circadian timing abilities of cells undergoing neural differentiation, neurospheres were prepared from mPer1::luc mouse SVZ and DG. Circadian bioluminescence rhythms were recorded in neurospheres maintained in culture medium that induces neurogenesis but not in one that maintains the stem cell state. Cell types were also characterized by confocal immunofluorescence microscopy at early and late developmental stages in vitro. Evidence was provided that neural stem progenitor cells (NSPCs) derived from the SVZ and DG of adult mice are self-sufficient clock cells capable of producing a circadian rhythm without input from known circadian pacemakers of the organism. Extremely rare percentages of mature neuronal cells were observed during ontogeny of rhythms. The bulk of the neurosphere cells were undifferentiated, indicating that they are the circadian clock cells producing timing signals. This conclusion was supported by immunocytochemistry for mPER1 protein that was localized to the inner, more stem cell-like neurosphere core. To further test whether circadian clocks in NSPCs are necessary for growth, differentiation and cell survival, neurospheres were cultured from Bmal1-/- and Cry1-/-,2-/- knockout mice. Neurosheres from Bmal1-/- knockout mice displayed unusually high differentiation into glia rather than neurons according to GFAP and NeuN expression, respectively, and very few DCX and BetaIII tubulin-positive, immature neurons were observed. The knockout neurospheres also had areas visibly devoid of cells and overall higher cell death. Neurospheres from mice lacking Cry1 and Cry2 showed significantly reduced growth. Altered NSPC proliferation and differentiation in these mice may impede memory formation and could provide a way to identify circadian timing effects in neurodegenerative disorders and impaired brain functions

Alternative CAIS analysis of freely moving mice imaged with photon counting mode.

Alternative CAIS analysis of freely moving mice imaged with photon counting mode.</p

Imaging chamber design.

A glass optical window rests on four 19 mm-long urethane foam spacers attached to the rim of the cylinder. A hole provides additional airflow and is also explored by the mouse as a nose-poke hole. A standard stainless steel sipping tube is attached and connected with a stopper to a 15-ml plastic centrifuge tube containing water or apple juice with luciferin. Ground corn cob bedding and mouse chow pellets are provided. (TIF)</p

Freely moving mice and unconscious mice imaged after intraperitoneal luciferin injection.

A: Brightfield image of an immobile (anesthetized) male Hr-Per1 mouse. B: Corresponding bioluminescence alone after 10-min image capture of the same mouse injected with luciferin showing high expression in several body areas including the submandibular gland (arrow) and testes. C-H: Bioluminescence in a freely moving male mouse after luciferin injection. Frames were selected to show various body postures captured during a 1-min sequence of 0.5 second images collected with 100x EM gain and 2 x 2-pixel binning. The entire body is visible and shows high gene expression in paws, ears, snout, and tail. Signal in the testes (C&H) is higher than the intensity range shown by the scale. The intensity ranges shown are in analog-to-digital units (ADUs) of the CCD camera.</p