Institute of Biology I, Animal Physiology and Neurobiology, University of Freiburg, Freiburg, Germany.
Gurunadh Reddy Chichili, Donatus Nohr, Michael Schäffer, Johannes von Lintig, Hans K. Biesalski; β-Carotene Conversion into Vitamin A in Human Retinal Pigment Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2005;46(10):3562-3569. doi: 10.1167/iovs.05-0089.
purpose. Vitamin A is essential for vision. The key step in the vitamin A biosynthetic pathway is the oxidative cleavage of β-carotene into retinal by the enzyme β,β-carotene-15,15′-monooxygenase (BCO). The purpose of the study was to investigate β-carotene metabolism and its effects on BCO expression in the human retinal pigment epithelial (RPE) cell line D407.
methods. BCO mRNA and protein expression were analyzed by real-time quantitative PCR and Western blot analysis, respectively. BCO activity was assayed in protein extracts isolated from D407 cells. The conversion of β-carotene to retinoids was determined by measuring retinol levels in D407 cells on β-carotene supplementation.
results. By RT-PCR, BCO mRNA was detected in D407 cells, bovine RPE, and retina. Western blot analyses revealed the presence of BCO at the protein level in D407 cells. Exogenous β-carotene application to D407 cells resulted in a concentration (75% at 0.5 μM and 96% at 5 μM; P < 0.05)- and time (127% at 2 hours and 97% at 4 hours in 5 μM β-carotene, P < 0.05)-dependent upregulation of BCO mRNA expression. Application of exogenous retinoic acid downregulated BCO mRNA levels at higher concentrations (1 μM; −96%, P < 0.0005) and upregulated it at a lower concentration (0.01 μM; 399%, P < 0.005). The RAR-a-specific antagonist upregulated BCO expression by sixfold (P < 0.005). Tests for enzymatic activity demonstrated that the mRNA upregulation resulted in enzymatically active BCO protein (7.3 ng all-trans-retinal/h per milligram of protein). Furthermore, D407 cells took up β-carotene in a time-dependent manner and converted it to retinol.
conclusions. The results suggest that BCO is expressed in the RPE and that β-carotene can be metabolized into retinol. β-Carotene cleavage in the RPE may be an alternative pathway that would ensure the retinoid supply of photoreceptor cells.
The eyes depend on blood circulation for vitamin A. Alternatively, vitamin A may be generated in the RPE from provitamin A carotenoids. The key step in vitamin A formation is the oxidative cleavage of provitamin A carotenoids by β,β-carotene 15,15′-monooxygenase (BCO), formerly known as β-carotene dioxygenase (EC 1.14.99.36). BCO cleaves β-carotene symmetrically into two molecules of all-trans-retinal. The activity of BCO was reported in cytosolic preparations of rat liver and intestine as early as the 1960s. 10 11 In recent years, the BCO gene was cloned from several vertebrate species, including humans. BCO is expressed in a wide range of human tissues, including the intestinal tract, liver, kidney, prostate, testis, ovary, and skeletal muscle. 12 It is also expressed in monkey retina and human retinal pigment epithelium (RPE). 13 Yan et al. 14 reported the human BCO gene structure and chromosomal localization and also demonstrated that BCO expression is higher in the RPE than in liver, kidney, intestine, or testes. 14 A Drosophila BCO mutant exhibits significantly reduced rhodopsin content and strongly reduced visual sensitivity that can be rescued exclusively by consumption of retinal, but not by consumption of β-carotene. 15 In zebrafish (Danio rerio), BCO plays a crucial role in the development of the eyes, the craniofacial skeleton, and the pectoral fin. 16 17 These findings suggest that β-carotene conversion by BCO may constitute an important step in the pathway leading to retinoic acid in local tissue environments of vertebrates.
In the present study, we elucidated the possible role of BCO in the conversion in the RPE of provitamin A, which plays an essential role in the retinoid metabolism of the eyes. We demonstrated BCO mRNA, protein, and enzyme activity in the human RPE cell line D407. Application of exogenous β-carotene to D407 cells resulted in an upregulation of BCO mRNA and protein levels, accompanied by a conversion of β-carotene to retinoids. Together, our analyses suggest that β-carotene cleavage, via BCO in the RPE, contributes to the function and survival of the adjacent photoreceptor cells.
All-trans-β-carotene in water-soluble beadlet form and the identical formulation without β-carotene (placebo) were generously provided by BASF (Ludwigshafen, Germany). All cell culture media and reagents were obtained from Biochrom (Berlin, Germany). The RAR-α antagonist Ro 41-5253 was a gift from Hoffmann-La Roche (Basel, Switzerland). All-trans-retinoic acid was purchased from Sigma-Aldrich (Munich, Germany).
The D407 (human RPE) cell line was a gift from Richard Hunt (Department of Pathology and Microbiology, University of South Carolina Medical School, Columbia, SC), 21 and the Caco-2 TC7 clone was kindly provided by Monique Rousset (INSERM U178, Unite de Recherches sur la Differenciation Cellulaire Intestinale, Villejuif cedex, France). 22 The D407 and Caco-2 TC7 cells were grown in DMEM supplemented with 10% FBS.
Water-soluble all-trans-β-carotene was dissolved in sterile distilled water by incubating at 37°C for 30 minutes and was stored at −70°C in aliquots. An aliquot of the suspension was extracted as described for the cells, and β-carotene concentration was determined by measuring the absorbance at λ = 450 nm in a UV/vis-spectrophotometer. When stored at −70°C, the β-carotene concentration was unchanged in the suspension for a period of 6 months. The cell cultures were maintained in serum-free medium for 48 hours before incubation in supplemented medium. All the preparations involving β-carotene were made under safe dim yellow light. The cells were incubated with all-trans-β-carotene-supplemented medium. Stocks of all-trans-retinoic acid and Ro 41-5253 (1 mM) were prepared in absolute ethanol and used for supplementation. The controls received the same amount of placebo (water-soluble formulation without β-carotene) or ethanol.
Total RNA was isolated from the cells in affinity columns (RNeasy; Qiagen, Hilden, Germany). DNase I (Qiagen) digestion was performed to avoid any DNA contamination. RNA was quantified in a spectrophotometer (Smartspec; Bio-Rad, Munich, Germany). Total RNA (2 μg) was reverse transcribed with an oligo(dT)20 primer (Thermo Hybaid, Ulm, Germany) using reverse transcriptase (Omniscript; Qiagen) according to the manufacturer's instructions.
Gene specific primers were designed from the respective gene mRNA sequences available in the GenBank sequence database (http://www.ncbi.nlm.nih.gov/Genbank; provided in the public domain by the National Center for Biotechnology Information, Bethesda, MD) using the Primer 3 program (http://www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgi) 23 available online. Primer sequences used: BCO (GenBank accession no. NM_017429) forward, 5′-CATCTTCCTTGAGCAGCCTTTC-3′, reverse 5′-GCAGCCGTCCTCTTCGTAG-3′; GAPDH (NM_002046) forward, 5′-ATGACATCAAGAAG GTGGTGA-3′, reverse, 5′-CTGTAGCCAAATTCGTTGTCA-3′; CYP26 A1 (NM_000783) forward, 5′-GGAGGACACGAAACCAC-3′, reverse, 5′-TCAGAGCAACCCGAAACC-3′; RAR-a (X56057) forward, 5′-GGGGAAGGAGTGTAGGATACC-3′, reverse 5′-CTGGGAAGGGCGAGTCTTA-3′; and RXR-a (NM_002957) forward, 5′-GCCTGAGTCTTCTCCTTGCT-3′, reverse, 5′-AGTTCCTGAGCCCCTCTCTC-3′. PCR amplification conditions were: 95°C for 4 minutes, followed by 35 cycles of amplification (95°C 30 seconds, 56°C 30 seconds, 72°C 30 seconds), followed by final an extension at 72°C for 5 minutes. The amplification reaction was performed in a thermal cycler (iCycler; Bio-Rad, Munich, Germany). The presence of the corresponding PCR products was determined by agarose gel electrophoresis.
The authenticity of the PCR amplified fragments was determined by DNA sequencing. The PCR products were cloned into the PCR II TOPO vector using a cloning kit (TOPO TA; Invitrogen, Karlsruhe, Germany). The plasmids containing amplicons were linearized with BamHI and used as standards for absolute quantification.
The cDNA from a 20-μL reverse transcription reaction was diluted to 100 μL with water and 5 μL was used for each PCR reaction. The amplification was performed with PCR master mix (QuantiTect SYBR Green; Qiagen) in triplicate reactions. The reaction mixture consisted of 50 μM of each forward and reverse primer, 1× master mix, and 10 nM fluorescein calibrating dye. The amplification was performed in a thermal cycler with an optical detection system (iCycler; Bio-Rad). The amplification conditions are 15 minutes at 94°C for the enzyme activation followed by 40 cycles of denaturation at 94°C for 30 seconds, primer annealing at 56°C for 30 seconds, and extension at 72°C for 30 seconds. The fluorescence data were collected at the extension step of each cycle. At the end of every PCR run, a melting curve was generated by increasing the temperature very slowly (0.5°C every 10 seconds) from 65°C to 95°C. The postrun data were analyzed with the real-time detection system software of the thermal cycler (iCycler iQ ver. 3.0; Bio-Rad). Briefly, the threshold value (C T) was selected in the log phase of the amplification curve. A standard curve (plot of C T values/crossing points of different standard dilutions against the log of amount of standard) was generated by using serial 10-fold dilutions (100–106 copies) of the standards with known quantities of DNA (linearized plasmid containing the PCR fragment). The copy numbers of the samples were deduced by plotting the C T values of unknown samples on the standard curve. Melting curve analysis was included in every run, to confirm the specificity of the PCR reaction. The changes in the mRNA levels were quantified by calculating the average of triplicate reactions and normalizing with mRNA levels of the GAPDH gene.
After the treatment, cells were washed with PBS, scraped into PBS, and sedimented by centrifugation. The cell pellets were dissolved in lysis buffer (1% NP-40, 0.1% SDS, 0.5% sodium deoxycholate, and protease inhibitors in PBS) and incubated on ice for 30 minutes. The lysates were centrifuged at 10,000g for 15 minutes at 4°C, and the supernatants were saved. Protein concentration was then determined (Dc Protein assay system; Bio-Rad). Protein from each preparation (100 μg) was separated by 10% SDS-PAGE, transferred onto a polyvinylidene difluoride (PVDF) membrane (Roche Diagnostics, Mannheim, Germany), blocked with 3% BSA, and probed with a murine BCO antiserum 16 overnight. Peroxidase-conjugated anti-mouse IgG was used as the secondary antibody. The blots were visualized by a chemiluminescence system (Cell Signaling Technology, Beverly, MA). Afterward, the detection blots were stripped and probed with actin antibody as an internal control.
Enzyme preparation and BCO activity assays were performed as described by During et al. 24 with minor modifications. Protein isolated from Caco-2 TC7 cells was used as a positive control in assay reactions. The reaction was initiated by the addition of 45 μL of 75 μM water-soluble β-carotene. The reaction mixture contained 40 μM tricine-KOH buffer (pH 8.0), 0.2 μM dithiothreitol (DTT), 1.6 μM sodium cholate, and 1 mg protein preparation from the cells. The enzyme reaction was performed at 37°C in a water bath with gentle shaking for 60 minutes and was stopped by adding 50 μL of 37% formaldehyde. The reaction mixture was extracted three times with n-hexane. Solvent was evaporated under a nitrogen gas stream, and the residues were dissolved in 100 μL acetonitrile and analyzed by reversed-phase HPLC. Results are expressed as nanograms of all-trans-retinal per milligram of protein per hour.
D407 cells were incubated with different concentrations of β-carotene. After the incubation, the cells were washed three times with phosphate-buffered saline (PBS), scraped with a cell scraper, and pelleted down by centrifuging at 1500g for 5 minutes. Vitamin A metabolites were extracted as described elsewhere. 25 Briefly, the cell pellet was dissolved in 100 μL of PBS, and the cells were lysed by three freeze–thaw cycles in liquid nitrogen. Ethanol (100 μL) was added, and the lysate was shaken with 2 mL of n-hexane for 30 minutes. The hexane phase was separated by centrifuging at 3000g for 1 minute and transferred into a fresh tube. The hexane extraction was repeated two times, and the phases were pooled and dried under a stream of nitrogen gas. The dried residues were reconstituted in 100 μL acetonitrile. A portion (50 μL) of the sample was injected into a reversed-phase C18 3-μm column with the dimensions of 300 × 4 mm (Nucleosil; Grom, Herrenberg, Germany). β-Carotene was eluted with an isocratic solvent containing 82% acetonitrile, 15% dioxane, and 3% methanol with 0.1% triethylamine at the flow rate of 1 mL/min and detected by UV absorption (λ = 450 nm). Retinol and retinal were eluted with the gradient of (A) acetonitrile/methanol (90/10, vol/vol) and (B) tetrahydrofuran, in the following program: 0 minutes: 98% A + 2% B; 6 minutes: 94% A + 6% B; 7 minutes: 80% A + 20% B; 16 minutes: 70% A + 30% B; 17 minutes: 98% A + 2% B; and 25 minutes: end of run and detected by UV absorption (λ = 325 nm). Chromatograms were recorded and analyzed (Star Chromatography Workstation Software, ver. 5.31; Varian Deutschland GmbH, Darmstadt, Germany).
The statistical significance at each time point, comparing the control with the treated conditions, was determined with a paired two-tailed Student's t-test. P < 0.05 was considered significant.
RT-PCR analysis of total RNA from the D407 cell line, Caco-2 TC7 cell line, bovine RPE/choroids, and retina yielded a PCR product of the expected size (237 bp), indicating that BCO mRNA is expressed in these cell lines and tissues (Fig. 1A) . Relative BCO expression, normalized with the GAPDH expression in corresponding tissues, is shown in Figure 1B . Caco-2 TC7 is a clone of Caco-2, which is a human colon cancer cell line. The TC7 clone has been reported to possess BCO activity 24 and was chosen as a standard for BCO expression.
Total protein was isolated from confluent cultures of D407 and Caco-2 TC7 cell lines. Protein extract (100 μg) was separated by SDS-PAGE, and BCO protein was detected by Western blot analyses. A single band of 65 kDa was detected in D407 cells and Caco-2 TC7 cells (Fig. 2) . The BCO antibody used in this study shows cross-reaction with a second member of the BCO gene family, β-carotene 9′,10′-oxygenase (BCO2). 26 However, BCO2 mRNA was not detectable in this cell line (Fig. 2B) , thus excluding the possibility that BCO2 protein was detected in the Western blot analysis, as well as BCO. In addition, BCO heterologously expressed in COS7 cells is specifically detected by the antiserum (Von Lintig J, personal communication, June 2004).
D407 cells were incubated with 0.5 and 5 μM β-carotene in the medium for 4 hours to investigate the effect of β-carotene on BCO expression. Upregulation of BCO mRNA was observed in the cells incubated with 0.5 μM (75%, P < 0.05) and 5 μM β-carotene (96%, P < 0.05) compared with control cells (Fig. 3A) . BCO protein expression increased 40% at concentrations of 0.5 and 5 μM, as judged by quantitative Western blot analysis (Fig. 3C) . To establish the time course of β-carotene action, we incubated cells in the medium supplemented with 5 μM β-carotene for 2, 4, 6, 24, and 48 hours. BCO mRNA was upregulated by 127% (P < 0.05) at 2 hours and 97% at 4 hours and was downregulated by 37% at 24 hours and 40% at 48 hours (Fig. 3B) . BCO protein expression had increased twofold at 2 and 4 hours but then decreased after 24 and 48 hours (Fig. 3C) .
CYP26A1 is an enzyme involved in retinoic acid catabolism, which converts retinoic acid into polar metabolites. CYP26A1 mRNA levels were measured in cells incubated with β-carotene, which had been used for BCO mRNA quantification. CYP26A1 mRNA levels were upregulated 44% at 0.5 μM and 70% at 5 μM after 4 hours of incubation (Fig. 3A) . In cells incubated with 5 μM β-carotene, CYP26A1 mRNA-concentration increased up to 10-fold with the incubation time and reached maximum peak levels (P < 0.05) after 48 hours (Fig. 3B) . This suggests that, from β-carotene, RA is synthesized in D407 cells, which subsequently induces CYP26A1 expression.
To elucidate the possible role of RA in the regulation of BCO expression, cells were incubated with all-trans-retinoic acid–supplemented medium for a fixed time of 6 hours. Subsequently, we analyzed BCO mRNA levels by real-time PCR. BCO expression was upregulated by 60% (P < 0.05) at 0.01 μM, 18% (P < 0.05) at 0.1 μM, and downregulated by 56% (P < 0.05) at 1 μM (Fig. 4A) . BCO protein expression was upregulated by onefold at 0.01 and 0.1 μM and downregulated at 1 μM (Fig 4C) . The cells were incubated with 0.01 μM retinoic acid for the indicated time points to establish the time course of the effect of retinoic acid on BCO expression. BCO mRNA levels were upregulated by 130% (P < 0.05), 196%, 250% (P < 0.005) and 399% at 2, 4, 6, and 24 hours, respectively, in cells incubated with 0.01 μM retinoic acid compared with the control (Fig. 4B) . BCO protein was upregulated over time with a maximum fivefold increase at 24 hours in 0.01 μM retinoic acid–incubated cells (Fig. 4D) . BCO mRNA and protein were decreased 10-fold in cells incubated with 1 μM retinoic acid (Figs. 4E 4G) .
To demonstrate the direct involvement of RA-signaling in the regulation of BCO mRNA expression, cells were incubated with the RAR-α-specific antagonist Ro 41-5253 at 1 μM concentration. After this treatment, BCO expression increased twofold at 2 hours (P < 0.005), threefold (P < 0.05) at 6 hours, and fivefold after 24 hours (P < 0.005; Fig. 4F ). BCO protein also increased with incubation time, to a maximum of fourfold at 24 hours (Fig. 4H) .
RAR-a and RXR-a mRNAs were detected in D407 cells. Expression of both mRNAs was strongly induced by retinoic acid in a concentration- and time-dependent manner (data not shown). β-Carotene incubation also induced RAR-a and RXR-a. RAR-a was induced by 50% at 0.5 μM, 80% at 5 μM, and 107% at 40 μM after 4 hours. RXR-a was induced by 50% in 0.5 and 5 μM and was unchanged in 40 μM β-carotene incubated cells (Fig. 5A) . In cells incubated with 5 μM β-carotene, RAR-a expression increased over the incubation time with a maximum of a 10-fold increase after 24 hours. RXR-a expression increased by fourfold at 2 hours and twofold at 4 hours and then declined to basal level at 6 hours and 24 hours (Fig. 5B) .
So far, we have shown that BCO is expressed at the mRNA and protein level. To demonstrate BCO at the functional level, we performed tests for enzymatic activity in total protein extracts from D407 cells and Caco-2 TC7 cells. Enzyme activity was observed by the formation of 4.5 ng retinal/h per milligram protein in D407 cells. In extracts from Caco-2 TC7 cells, 7.3 ng retinal/h per milligram protein was found (Fig. 6) . All-trans-retinal was the only product detected in the in vitro test system.
Cells were incubated in medium supplemented with 5 μM β-carotene, to determine its uptake and metabolism. Cells took up β-carotene, but the uptake rate slowed with the incubation time (Table 1) . In addition, we performed HPLC profiling for retinol and retinal at different time points. As shown in the Table 1 , retinol was present in cells incubated with 5 μM β-carotene and slightly increased with incubation time, whereas no retinol was found in the control cells or at the starting point of β-carotene incubation. Retinal was below the detection limit in both control and β-carotene-incubated cells, which may have been due to the rapid metabolic conversion to retinol in the cells.
In humans, provitamin A carotenoids are the major dietary precursors for vitamin A. In the photoreceptor outer segments, rhodopsin exists in millimolar concentrations. Therefore, eyes have a very high demand for vitamin A, to maintain the visual process. Bernstein et al. 28 reported the presence of β-carotene in the human RPE/choroid (10 ng/0.2 g tissue). Recently, Yan et al. 12 demonstrated that BCO expression in the RPE is higher than in liver, kidney, intestine, and testes. In the present study, we investigated provitamin A metabolism in the human RPE cell line D407. This cell line is an immortal human RPE line that retains many of the metabolic and morphologic characteristics of RPE cells found in vivo, even after several passages, and it has a long survival time. 21 In the present study, exogenous β-carotene application to D407 cells resulted in an upregulation of BCO mRNA and protein levels in a time- and concentration-dependent manner, resulting in the conversion of β-carotene to retinol.
Furthermore, β-carotene incubation resulted in an upregulation of CYP26A1 mRNA in a concentration- and time-dependent manner. CYP26A1 is known to be induced by retinoic acid and its receptors through a transactivation of a retinoic acid–responsive element (RARE) located in its promoter. 29 This observation indicates a possible involvement of retinoic acid in the regulation of β-carotene metabolism in the RPE. Indeed, BCO mRNA and protein levels first increased in D407 after β-carotene supplementation, but decreased after longer incubation periods. The primary product of β-carotene cleavage via BCO is retinal, which can be further metabolized either to retinol or retinoic acid. In D407 cells, we found significant amounts of retinol, but retinoic acid was below the detection limit during the initial periods and was detectable only after 24 hours. Nevertheless, traces of retinoic acid are sufficient to activate its nuclear receptors. In addition, an increase in cellular retinoic acid levels rapidly induces its catabolism through CYP26 monooxygenases. Thus, the strong induction of CYP26A1 at the mRNA level and the downregulation of BCO after prolonged β-carotene supplementation may be caused by the increase of retinoic acid in D407 cells after β-carotene supplementation. Retinoic acid's effects are mediated through its nuclear receptors, RAR and RXR. 30 31 We showed that RAR-a and RXR-a are expressed in D407 cells, and their expression is enhanced by β-carotene.
In a recent report, peroxisome proliferator-activated receptor (PPAR)-gamma/RXR-a heterodimers were shown to regulate BCO gene expression, and 9-cis-retinoic acid treatment induced the expression in mice. 33 In our study, neither 9-cis-retinoic acid nor 9-cis-β-carotene was detected in the cells after β-carotene incubation. Therefore, a role of 9-cis-retinoic acid in the observed effects on the regulation of BCO expression in D407 cells can most probably be excluded. We also performed a transcription factor binding site search in the 5′ flanking region of the human BCO gene (GenBank sequence database) that revealed a putative RARE (GGGTCActtgAGGTCA, 600 bases upstream to the start site; Chichili GR, unpublished observation, 2004). Therefore, we assume that RAR-a and RXR-a are involved in the regulation of BCO expression in D407 cells and thus most probably are also involved in vivo in the RPE. Bachman et al., 34 demonstrated in rats and chicken that retinoic acid feeding results in a decrease in BCO enzyme activity in intestines but did not affect BCO activity in liver. Our data partially agree with the observation that high RA concentrations inhibit BCO expression. However, we also found that low RA levels have a positive effect on BCO mRNA expression. The fact that RA induces BCO expression at a lower concentration may contribute to efficient the use of β-carotene for retinoid synthesis, whereas the suppression of BCO expression at higher RA concentration may avoid an excess of retinoid synthesis from its provitamin A precursor. Further investigations are necessary to gain a mechanistic understanding of the regulation of BCO expression via RA and its nuclear receptors in different cell types and organisms.
Bhatti et al., 13 reported that BCO is expressed at variable levels in the RPE of different species but is not expressed in RPE cell lines. This disparity may be due to the sensitivity of the RT-PCR and RNA purification used in our study. RNA isolated with affinity columns was used for RT-PCR in our study, which works better than RNA isolated by phenol-based methods (data not shown). Primers with high PCR efficiency derived from the human BCO sequence were used in the present study. This may explain the differences between the two studies, because PCR sensitivity varies between different primer sets. In addition, our mRNA expression data in RPE cells are further supported by assessing BCO protein levels as well as by tests for BCO enzymatic activity.
Recently, expression of BCO has been reported in epithelia of several tissues in human including glandular cells in the prostate, endometrium, mammary tissue, kidney tubules, keratinocytes of the squamous epithelium of skin; steroidogenic cells in testis, ovary, and adrenal gland; and skeletal muscle cells. 35 Epithelia in general are structures that are very sensitive to vitamin A deficiency. Local β-carotene cleavage may contribute to the maintenance of a steady state level of retinoids in these tissues, particularly under conditions of high retinoid demand. BCO mRNA expression was also found in bovine RPE and retina, indicating a possible role of this enzyme in the vitamin A metabolism of other species. Thus, our study supports previous findings 14 28 and provides functional evidence that β-carotene conversion to vitamin A exists in the RPE and may contribute to ensure retinoid-dependent processes in vision.
A most interesting finding in two siblings with impaired retinol transport caused by mutations in the RBP gene was that only mild clinical vitamin A deficiency symptoms such as night blindness and modest retinal dystrophy were present. 36 In these patients, cleavage of β-carotene in the RPE may compensate for the impairments in retinol transport. In such situations, high doses of β-carotene supplementation may help to avoid vitamin A deficiency.
More generally, β-carotene is the major source to ensure the vitamin A demand of humans. Yet, it was thought that β-carotene is converted to vitamin A immediately after its uptake in the small intestine and stored in the liver in the form of retinyl esters, implying that peripheral tissues mainly rely on the supply of preformed vitamin A in the circulation. Our study provides evidence for a tissue-specific vitamin A synthesis in the RPE. This finding is in agreement with the fact that β-carotene is transported in lipoproteins and BCO is expressed in various human tissues.
Taken together, the evidence in this study showed that BCO is expressed and can be induced by β-carotene in RPE cells. In addition, β-carotene can be converted into vitamin A in RPE cells in a regulated manner, thus providing functional evidence for an eye-specific provitamin A metabolism in humans.
3 Department of Biochemistry, Meharry Medical College, Nashville, Tennessee.
Supported by The European Union (ProVitA).
Submitted for publication January 25, 2005; revised May 24, 2005; accepted August 25, 2005.
Corresponding author: Hans K. Biesalski, Institute of Biological Chemistry and Nutrition (140), Garbenstrasse 30, University of Hohenheim, D-70599 Stuttgart, Germany; biesal@uni-hohenheim.de.
BCO mRNA expression in RPE cell lines and bovine retinal tissues. Total RNA from confluent cell cultures and bovine tissues was extracted, 2 μg of total RNA was reverse transcribed into cDNA, and PCR amplification was performed with human BCO gene–specific primers. PCR amplicons were separated by electrophoresis on a 1% agarose gel, stained with ethidium bromide, and documented in a UV gel documentation system. (A) Lane M: 100-bp ladder size marker; lane 1: Caco-2 TC7 cells; lane 2: D407 (human RPE cell line); lane 3: bovine RPE; and lane 4: bovine retina. GAPDH expression in corresponding samples is shown. (B) BCO mRNA levels were quantified by real time quantitative PCR and normalized with GAPDH mRNA expression in corresponding samples. The mean ± SE of two separate experiments is shown.
(A) BCO protein expression in D407 and Caco-2 TC7 cell lines. Protein extracted from confluent untreated cell cultures was analyzed for BCO protein expression by Western blot analysis. Lane +: Positive control (BCO fusion protein with an N-terminal His6-tag); lane 1: D407 cells; and lane 2: Caco-2 TC7 cells. (B) BCO II mRNA expression. Total RNA was extracted from confluent cell cultures, 2 μg of total RNA was reverse transcribed into cDNA, and PCR amplification was performed with human BCO II gene–specific primers. PCR amplicons were separated by electrophoresis on a 1% agarose gel, stained with ethidium bromide, and documented in a UV gel documentation system. Lane M : 100-bp size marker, BCO II; lane 1: negative control; lane 2: D407; lane 3: positive control, GAPH; lane 4: negative control; lane 5: D407; lane 6: positive control.
Concentration- and time-dependent regulation of BCO and CYP26A1 expression by β-carotene. (A) D407 cells were incubated in the medium supplemented with 0.5 and 5 of all-trans-β-carotene for 4 hours. (B) D407 cells were incubated in the medium supplemented with 5 μM β-carotene for 2, 4, 6, 24, and 48 hours. Total RNA was extracted and the changes in mRNA levels of BCO and CYP26A1 were quantified by real-time quantitative PCR. The mean ± SE of results in three separate experiments is shown except when the SE is too small to be seen. *P < 0.05) significant difference from the control. (C) Top: 100 μg of protein extracts from D407 cells treated with β-carotene was analyzed for BCO protein expression indicated the time points by Western blot analysis. Bottom: actin expression in the corresponding samples in the same blot.
Concentration- and time-dependent regulation of BCO expression by all-trans-retinoic acid. (A) D407 cells were incubated in the medium supplemented with 0.01, 0.1, and 1 μM all-trans-retinoic acid for 6 hours and (B) in medium supplemented with 0.01 μM all-trans-retinoic acid for 2, 4, 6, and 24 hours. (E) All-trans-retinoic acid (1 μM). (F) The RAR-a antagonist (1 μM). Total RNA was extracted, and the changes in the BCO mRNA levels were quantified by real-time quantitative PCR. The mean ± SE of results in three separate experiments is shown except when the SE is too small to be seen. **P < 0.005; *P < 0.05, significant difference from the control. BCO protein expression: D407 cells were incubated with (C) different concentrations or (D) 0.01 μM or (G) 1 μM all-trans-retinoic acid for the indicated times. (H) RAR-a antagonist (1 μM Ro 41-5253). BCO protein expression was analyzed by Western blot analysis Bottom: actin expression in corresponding samples in the same blot.
Regulation of RAR-a and RXR-a mRNA expression by β-carotene. (A) D407 cells were incubated in the medium supplemented with 0.5 and 5 μM of all-trans-β-carotene for 4 hours. (B) D407 cells were incubated in the medium supplemented with 5 μM β-carotene for 2, 4, 6, and 24 hours. Total RNA was extracted, and the changes in mRNA levels of RAR-a and RXR-a were quantified by real-time quantitative PCR. The mean ± SE of results in three separate experiments is shown, except when the SE is too small to be seen.
BCO enzymatic activity in D407 and Caco-2 TC7 cells. Protein extraction and a BCO activity assay were performed. Retinal produced from β-carotene in the presence of protein was quantified by HPLC. The mean ± SE of results in three separate experiments is shown.
D407 cells were incubated in medium supplemented with 5 μM of β-carotene for indicated time periods. β-Carotene and retinol were extracted and analyzed by HPLC as described in the experimental procedures. The mean ± SE of two separate experiments is shown.
The authors thank Christian Köpsel (BASF) for providing water-soluble β-carotene, Willi Hunziker (Hoffmann-la Roche) for providing the Ro 41-5253; Andrea Flaccus for assistance with the PCR assays and Michael Wolter for assistance with the HPLC analysis.
RozenS, SkaletskyHJ. Primer3 on the WWW for general users and for biologist programmers.KrawetzS MisenerS eds. Bioinformatics Methods and Protocols: Methods in Molecular Biology. 2000;365–386.Humana Press Totowa, NJ.


Free resources for lecturers, educational ideas and ideas, instructional subjects, Private Studying Networks, Mission Primarily based Learning, Google, Evernote, Discovery Schooling and more. About Weblog – Math Weblog by David Sladkey. I educate High Faculty Arithmetic and I love my job. I wish to learn new things to develop into a better instructor. It is a record of my journey. Rooster Littling is about creating a way of urgency. It's not as common as it once was; chances are you'll recall that charter followers used to say that college students could not wait another day for public colleges, till they decided that constitution faculties wanted a number of more days to stand up to speed. Tereza Lombardi has over 30 years of expertise in instructing all levels and ages and in training academics. Her present job as a trainer in an progressive bilingual faculty, raised her interest in developing analysis in bilingualism and trainer training. She has MA in Applied Linguistics and is CELTA qualified.
Teachers who're in peril commuting don't have to come in. Academics who aren't, are available. These teachers who are available receive comp time within the kind of some further prep durations. Good principals understand how to try this at no expense to the college i.e. field trip, meeting, and so on. and so on. There are many different price impartial choices as effectively, but that is fodder for another publish. iii Given the size of the county, some black students, particularly these within the jap facet of the county, needed to endure two-hour bus rides to school, in dilapidated and segregated buses—or not go in any respect. Gandy High Alumni Reflect on College Days," Herald-Progress, September 10, 2009, p. 1.
In EXTRA's original settlement with NA, the 7 HS seats have been split 4-three with MORE getting the additional seat – New Action proposed that since MORE had gotten extra votes in the 2013 election. MORE selected its four individuals and likewise an alternate. Within a couple of days of beginning the petition campaign, one of the NA individuals pulled out and we had to fill this position ASAP – and we had an issue with our alternate who a few of us believed was not going to stay in instructing – and thus we'd lose that seat if we won.
Below a constitution-alternative system, schools do compete for college kids. However in such a system, all college students will not be created equal. Not all college students are equally desirable to charter schools. College students who don't include any further costs to coach (no special wants, no English-as-second-language issues, no behavioral issues) are desirable. College students who make good take a look at numbers for the school are desirable. Students who come from supportive dwelling environments are desirable. Students who cost extra, have more issues, have fewer skills, draw decrease scores- these college students usually are not as desirable to charters.
Democracies are constructed upon the rules of equality and the participation of citizens in public deliberation and determination making. However participation can solely work if folks have a minimum of a fundamental understanding of the system's norms and establishments, can kind opinions of their very own and respect those of others, and are keen to interact in public life a method or one other. A brand new Tendencies Shaping Training Highlight seems to be at how civic training can support college students in developing the information and abilities wanted to participate in the democratic process, especially in an increasingly digitalised world.


text:
Rolls-Royce says demand for luxury cars is recovering
Torsten Muller-Otvos said the demand meant Rolls-Royce Holdings Plc was the first car company to resume car production in the United Kingdom on May 4.
Reuters Published September 1, 2020
LONDON: The chief executive of Rolls-Royce said demand for his company's luxury cars is rebounding, helped by sales in Asia, and he is optimistic about the outlook for next year after the coronavirus pandemic hit consumer confidence and closed dealerships.
"We see a very fruitful business now coming back from Asia, also Europe is coming back on track, the Americas just delivered an excellent July result and August result," the boss of the BMW-owned Rolls-Royce Motor Cars told Reuters.
"I am quite optimistic looking into 2021, particularly on the back of a very strong order bank we have already on our books." Muller-Otvos said his company has no plans to move production out of the United Kingdom because of Brexit.
"We are committed to Britain. I would even call us being part of the British industrial crown jewels," he said. "For that reason, Rolls-Royce belongs to Britain."

redpajama_set_name: RedPajamaCommonCrawl


During my stay in Cebu, Philippines I have been couch surfing with some locals. Locals who not only turned out to be business owners, but also locals who turned out to be friends.
Grace, Verge, Phillip and Charlie are all founders and owners of Lupad Travel. A travel agency with a lot of opportunities to simplify your holidays as they handle the booking and all you have to handle is packing and getting on a flight to discover new adventures!
Couch surfing with these guys has been unbelievable, not only are they friendly and very competent within tour guiding; their hospitality exceeded every possible imagination.
We didn't book through the business Lupad Travel, but we did meet the people behind this company - and they're are absolutely amazing people. City tour in Cebu, snorkeling in Moalboal, taking dips at Kawasan Falls, visiting the monkeys at Leyte and spending a night at Canigao is just some of the amazing things we did with these guys! And as we left them to go on our own adventures here on this island and the surrounding islands, they helped out by planning and making our trip the best possible. They showed us the ways of the Philippines, from eating exotic food to using the local transportation to get around.
I wouldn't hesitate a second to recommend Lupad Travel to anyone I know. While traveling I have learnt that the locals always knows best. Where to eat, what to see and where to go - as goes for the Lupad Travel guys - Grace, Verge, Phillip and Charlie - they definitely know exactly how to plan your perfect holiday.


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Candy IndustryCandy Industry News
Corn syrup name debate continues
District Court Judge Marshall adds Cargill, ADM as defendants in "corn sugar" case.
By Alyse Thompson
Depending on which side you're on, the battle over what to call high fructose corn syrup has either sweetened — or soured.
Los Angeles U.S. District Court Judge Consuelo Marshall ruled Wednesday that sugar refiners such as Cargill and Archer-Daniels-Midland (ADM) can be included in the April 2011 lawsuit filed by the Sugar Association, the Western Sugar Cooperative (WSC) and others against the Corn Refiners Association (CRA).
The lawsuit, the WSC says, is an attempt to keep the CRA and other large sugar refiners from labeling high fructose corn syrup as simply "corn sugar" for fear of misleading consumers. The CRA, meanwhile, says the label does not have to be so specific, as the human body cannot tell the difference between high fructose corn syrup and other sugars.
Nevertheless, the WSC says it is happy to see the focus turn to large sugar refiners, citing Marshall's ruling that "plaintiffs allege with particularity facts for a false advertising claim against member companies ADM, Cargill, Corn Products, and Tate & Lyle."
Additionally, sugar farmer attorney Adam Fox says increasing the scope of the suit could help millions of American corn product consumers.
"Judge Marshall's ruling clears the way to allow this lawsuit to proceed so that we can assure an end to the false advertising and make the agribusiness giants behind it answer for their misconduct," Fox says. "We look forward to taking the next steps in this important case."
The CRA and involved sugar refiners are not as pleased, however.
"This ruling is solely about who is included in the lawsuit and has no bearing on the merits of the case which are about ensuring that consumers get the facts regarding high fructose corn syrup," says Audrae Erickson, CRA president."
The CRA believes Marshall made the only ruling pertaining to the case in a preliminary review in October, citing her statement that the sugar associations and farmers "have not presented any evidence to support their burden on the claims that CRA's statements have influenced any purchasing decisions [or] that plaintiffs have suffered any injury."
Additionally, the CRA says it has the consumer in mind while it make its arguments, similar to the WSC and the Sugar Association.
"Consumers have a right to know what ingredients are in their food and beverages, especially as we're seeing more and more people concerned with managing their sugar intake," Erickson says. "It's essential for consumers to understand that high fructose corn syrup is another kind of sugar despite the processed sugar industry's attempts to censor our education campaign."
Nonetheless, the Food and Drug Administration recently rejected the CRA's petition to change the name of high fructose corn syrup because it "would not accurately identify or describe the basic nature of the food or its characterizing properties."
A date for the current trial has not been set, but when it happens, conflicting testimonies will likely make deciding the case a bit sticky.
KEYWORDS: Corn Refiners Association high fructose corn syrup name dispute Sugar Association
Alyse Thompson has held many positions with BNP Media, first serving as an intern at Candy Industry Magazine in summer 2012. She joined Candy Industry's staff full time as associate editor in August 2016 after a few years at newspapers in West-Central Illinois, becoming managing editor in March 2019. Alyse has also served as managing editor of Cannabis Products Magazine since March 2019. She has bachelor's degrees in journalism and Spanish from Western Illinois University.
Does corn syrup by any other name taste as sweet?
Suit targets high fructose corn syrup 'sugar' name claim
Report: China's confectionery market continues to grow
Cargill is committed to helping customers navigate today's complex marketplace, using our unmatched ingredient portfolio and extensive technical expertise to solve the most complex product demands. By collaborating with our team of experts, we can help you create confectionery products that deliver on product goals.
Zachary Confections Inc.
"A Family Owned American Company" Since 1950.
Ciranda supplies organic, non-GMO and fair-trade ingredients with expertise in rice and tapioca syrups, syrup solids and maltodextrin; agave syrup; honey; cocoa powder; chocolates; coconut products; liquid and powdered lecithin; RSPO-IP sustainable palm oils and more. Request samples at www.ciranda.com.


Lunches are brought to CRCA by the Nutrition Group, the food service at Uniontown High School. We also have deliveries from Papa John's Pizza on pizza days. Milk is served with each lunch. The cost for lunch is $3.25 and is payable when your lunch is ordered each day.


Pinkman White [Bullion Foundry] is an Australian owned and operated company, specialising in helping everyday buyers to get as close as possible to the source of bullion – where the best prices can be found.
Our humble roots in trading have provided us with an understanding of local markets. With an extensive network of bullion suppliers and a commitment to continually seeking out the best possible price, our mission is to pass those prices to our members in the most simple and risk-free way possible.
Bullion Foundry, a trading style of Pinkman White, provides retail investors an opportunity to buy precious metals at the same beneficial rate as large-scale investors through what is effectively group purchasing via a maximum-bid process. Unique within the retail bullion market, Bullion Foundry is part investment portal, part buyers club.
When doing business with Bullion Foundry - please say you saw them in Bullion.Directory.
Read or add Bullion Foundry reviews and ratings using the "reviews" tab above.
All Bullion Foundry details are believed to be correct at the time of listing. Please report errors here.


You will be assessed by an independent trainer assessor at the end of the course.
Candidate must be able to swim 50m in less than 60 sec.
36 hours of training needs to be completed by the candidate before going forward for assessment.
Swimwear, shorts and T-shirts is needed for the duration of the course.
Cost: €390 including all learning materials, assessment, logbook & 2 year qualification.
Renewal candidates also welcome on these courses.
The renewal component of this course is made up of 20 hours at a cost of €200.


A dummy plug device that connects to any HDMI® output and emulates the presence of an HD 1080p60 display device. Sold exclusively as part of a selection of Crestron Flex UC system kits.
Provides a dummy plug device that connects to any HDMI® output and emulates the presence of an HD 1080p60 display device. It is intended for use with the Crestron® UC Engine (UC-ENGINE) as part of a complete Crestron Flex UC system.
This product is sold exclusively as part of a complete Crestron Flex system kit.
This product is sold only as part of a complete system kit, which may be purchased from select authorized Crestron dealers and distributors. To find a dealer or distributor, please contact the Crestron sales representative for your area. A list of sales representatives is available online at https://www.crestron.com/How-To-Buy/Find-a-Representative or by calling 855-263-8754.
Crestron and the Crestron logo are either trademarks or registered trademarks of Crestron Electronics, Inc. in the United States and/or other countries. HDMI is either a trademark or registered trademark of HDMI Licensing LLC in the United States and/or other countries. Other trademarks, registered trademarks, and trade names may be used in this document to refer to either the entities claiming the marks and names or their products. Crestron disclaims any proprietary interest in the marks and names of others. Crestron is not responsible for errors in typography or photography. Specifications are subject to change without notice. ©2018 Crestron Electronics, Inc.


It's officially summertime and we are bringing you the anthems with "Songs Of Summer 2014." As long as skies are blue you'll want to blast this 133-143 BPM mix during every single workout. Featuring songs like "Wiggle," "Rude," and the massive hit "Summer," this album will have you feeling SO fancy. Turn down the HEAT for what?!


text:
Q: fsolve returns a point at which the function is not zero I'm quite new in Python programming and I'm struggling to optimize a simple non linear function. The code is as follows (I have anaconda on my computer which includes numpy and scipy libraries):
#!/usr/bin/python
# -*- coding: Latin-1 -*-

# System libraries

import os
import sys
import math
import getopt
import scipy

from scipy import optimize

#params
a = 78.0
b = 47.0
c = 0.0203242205945909

#initial guess (solution is not far to c so we use c parameter as initial guess)
guess = c
#objective function to find root
def obj(_x, _a, _b, _c):
    f = (_c - (_x + (_b*x**_b)*(10**_a)))
    return f;

x = guess

sol = scipy.optimize.fsolve(lambda x: obj(x, _a=a, _b=b,  _c=c), guess)
#aaaand when we calculate the solution, it does not correspond to a root of the function!
print(sol)
print(obj(x, a, b, c))

Result when I use script is:
[-1.38815731]
-1.40848153538

I would expect:
[correct root that I don't know]
0.00000000000

Why does it give that result? What did I do wrong? How to make fsolve verbose? I don't get that in documentation.

redpajama_set_name: RedPajamaStackExchange


text:
We've changed the talent game for innovation leaders
In 2014, clients and talent told us they wanted a new recruitment model - a model that respects innovation, growth and human excellence.
We call ourselves human capital developers where we align with innovation-leading employers and talent to provide the highest level of executive search and coaching services to move their vision forward.
We are a dedicated team of senior search headhunters and certified executive
coaches for global innovation leaders.
The headhunting team spends every day dismantling the old recruitment model to deliver the vision you demand. We work on building your employer brand to create a best-in-class candidate experience.
The certified executive coaching team understands the leadership challenges innovation leaders face every day to reach your career, company or market-making vision.
Caroline Stokes
Human Capital Developer and Certified Executive Coach
Caroline spent 20+ years with innovation technology leaders (PlayStation, Virgin, Nokia) critiquing hiring practices, onboarding practices, engagement and leadership systems. She spent an additional five years working at a headhunting firm delivering executive-level placements at Microsoft, Bethesda, Ziff Davis and Trinity Mirror.
Caroline realized something was missing from the process: a high-touch, compassionate approach that left both the employer and talent closer to their seemingly impossible future.
In 2014, Caroline founded FORWARD with a vision to create a constantly evolving human capital solutions company, with an ethical and humanist headhunting process and executive coaching practice for innovation leaders - so everyone can move forward.
Coaches & Associates
Chiara D'Avanzo
CHRP, M.Sc., Talent Acquisition Strategy Consultant
Chiara is a talent acquisition expert with 12 years of progressive experience helping top employers attract and hire talent, mainly in technology and healthcare sectors. In her view, recruiting talent doesn't have to be painful.
She partners with businesses that are creating workplaces for the workforce of tomorrow. She also enjoys taking outdated recruitment models and transforming them into the new era. Her approach is straightforward, highly collaborative and effective. Chiara will partner with you to figure out the best ways to win talent for your company's specific needs.
Recruiting highly specialized talent has become one of the most complex activities for businesses today and the status quo just doesn't work any more.
Miki Hara
Talent Acquisition & HR Strategy Consultant
Based in London, Miki Hara has been building high-performing teams in the Animation, Games, and Visual FX industries for 18 years.
She has recruited, trained and led creative and technical teams as a graphic artist, and for the past 10 years has mentored, coached and developed Senior Leaders, Producers, Supervisors, Artists, and Engineers as a deeply involved and committed HR & Recruitment leader.
Miki has expertly guided the development of great company cultures by ensuring that candidate and employee experience are held as top priorities at all levels of the organization from first contact, through short and long-term collaboration, to last day and beyond. The relationships she builds with her teams are lifelong and deeply important to her.
Amy Foley
Talent Research & Sourcing
Soon after FORWARD started in 2013, Amy Foley worked as our #1 undercover sourcer for international talent across VFX, Cloud, Games and Mobile.
With an analytical mind, a background in QA and a service-oriented virtual assistant style of working (which means everything happens right now), Amy performs fast and accurate talent searches to create critical pipelines that shorten the talent analysis and presentation process.
Amy loves the variety of impossible searches at FORWARD and finding the right candidates to ensure talent and organization fit, hand in glove.
Nicole Gravagna, PhD
Dr. Nicole Gravagna is a neuroscientist and executive coach. She is the president of NeuroEQ, a management consulting and change management firm. You may know her as the author of MindSET Your Manners or coauthor of Venture Capital for Dummies. She is a contributor to Forbes, Inc., Newsweek, Medical Daily, HuffPost UK, and a three-year Quora Top writer.
After coaching literally hundreds of startup CEOs through the process of raising venture capital, she noticed that money was only a symptom of the real problem. When CEOs were having trouble reaching their goals, the problem was inside them, usually in the form of limiting beliefs.
Now, Nicole coaches executives in huge and small companies alike. She uses neuroscience and behaviour data to inspire successful people to turn their persistent problems into permanent solutions often resulting in exceptional performance.
Rachel Nixon
Digital and Creative Advisor
Rachel is an award-winning digital leader who has shaped the strategic vision for global media brands, honed through two decades' working online.
She brings a unique combination of international editorial and executive experience at public media organizations – the BBC and CBC – and technology and business expertise from her role as Editor-in-Chief for News and Sports at Microsoft in the US.
Rachel has also held key roles in boundary-breaking journalism startups and recently graduated with honours from the professional photography program at the Vancouver Institute of Media Arts.
Whether fine art photography, or digital media brands, Rachel loves being able to use her storytelling skills to build strong connections with others.
We turn impossible tasks into reality through collaboration, hard detective work, and a deep understanding of innovation leadership to achieve success.
Are you ready to move FORWARD? We've got the talent aquisition and leadership solutions you need.


text:
Spotlight: Temptation by Fire by Tiffany Allee
Temptation by Fire
by Tiffany Allee
Visions of death haunt her . . .
Ava Keller sees the future. When she suffers a horrifying vision of a man's violent death, she knows from experience nothing she does will stop it from becoming a reality. But when she meets a strong, broken man, he makes her want to believe in second chances.
Vengeance consumes him . . .
Tortured by his past, Karson has worked tirelessly to wreak vengeance on the demons who murdered his family. But when Ava inadvertently disrupts his intricate plan for revenge, his focus shifts to an all-consuming need to keep her alive and in his arms.
Love could save their souls . . .
Thrown together by fate, Ava and Karson explore their new feelings and discover hope for the future. But they must battle the demons threatening humanity to find a destiny worthy of their love.
"So who are these Venators? Are they like some sort of demon-hunting organization?" I asked.
"They're like a need-to-know organization," Karson said.
"I haven't held anything back from you."
He shrugged and his hand dropped from my shoulder. "I'm trying to help you. That doesn't give you the right to know everything about me."
I opened my mouth to argue but he held a hand up, and his index finger touched my lips. The gesture was meant to shut me up, and it worked. In that moment every thought flew from my head. There was only him. The intense look in his eyes. The roughness of his skin on my mouth.
The immediate need that curled in my stomach.
What would it be like to be with a man like him? Rough and wild and freaking amazing, that was my bet. My body ached to find out. The rest of me knew better.
"You need to hide for a while. Then you can go back to your normal life. Promise me."
With a quick jerk of my head, I promised.
Tension flowed out of him visibly, and a small smile touched his lips. "Good."
And if I hadn't already convinced myself that he was probably right, that laying low couldn't hurt, his next words would have.
"Please don't go back on your promise. Don't make me watch them kill you."
CPA-turned-romance-author Tiffany Allee used to battle spreadsheets in Corporate America, and now concentrates on her characters' battles to find love. Raised in small-town Colorado, Tiffany currently lives in Phoenix, AZ, by way of Chicago and Denver. She is happily married to a secret romantic who tolerates her crazy mutterings.
She writes about ass-kicking heroines and the strong heroes who love them. Her work includes the suspense-driven From the Files of the Otherworlder Enforcement Agency series which revolves around a group of paranormal cops solving crimes and finding love, and Don't Bite the Bridesmaid, a lighthearted paranormal romance (Entangled Publishing).
Tiffany has an MBA in accounting and nearly a decade of experience in corporate finance. All super useful stuff for a writer who spends far too much time trying to figure out fun ways to keep her characters apart, and interesting ways to kill people (for her books—of course!).
You can find her at
Sign up for her newsletter here:
http://tiffanyallee.com/about/newsletter-sign-up/
$10 Amazon gift card, and an ebook copy of one of Tiffany's backlist books
By Unknown at 12:00 AM
Filed under: Bewitching tours, Temptation by Fire, Tiffany Allee


text:
Губкино — название населённых пунктов в России.

 Губкино — деревня в Новомосковском административном округе Москвы.
 Губкино — деревня в Одинцовском районе Московской области.
 Губкино — село в Малоархангельском районе Орловской области.
 Губкино — деревня в Новоржевском районе Псковской области.
 Губкино — село в Спасском районе Рязанской области.

См. также
 Губино
 Губкин
 Губкинский

redpajama_set_name: RedPajamaWikipedia


As Podcasts Continue To Grow, Who Listens Most?
It appears that podcasts are becoming more than an industry buzzword. A new study finds that 24% of those surveyed have now tuned into the audio platform, while a formidable 15% listen either daily or weekly.
When asked how often they tune in, 7% said daily, 8% said weekly and 4% said monthly. Another 5% responded several times a year, with 73% "rarely or never" accessing podcasts. "Given the portable nature of a podcast, it's possible that those who listen more regularly have found a way to incorporate their favorite podcasts into their daily or weekly routine, perhaps during a commute or while working out. The data supports this, as it shows daily podcast listeners do like to multitask with their technology," the report says.
The data also shows that men take a slight lead in those who listen more frequently, outweighing daily and weekly women listeners.


Once I heard a pastor say that in life's relationships, after a while, some people suffer from a condition known as 'See- finish-ism'.
It's a condition in which, because of prolonged contact with someone, or because you literally know them inside out-having lived with them or worked with them-they sort of lose the hype you had about them at initial contact. You know their faults, weaknesses, challenges. All of a sudden, they don't seem very special anymore; they become like everyday people.
It happens everyday-spouses that have 'seen their spouses finish', congregations that have 'seen their pastors finish', even children that have 'seen their parents finish'. All of a sudden, these people have very little worth in your eyes. You don't even want to hear anything they have to say. 'What new thing do they want to tell me that I have not heard before?' you ask.
I think that this was probably the condition that the people of Jesus' hometown must have suffered from. This was Jesus, who grew up with them in their town. They'd seen him as a child. They knew his roots. Listen to the sneer in their voices.
"…Where did this man get this wisdom and these mighty works? Is not this the carpenter's son? Is not his mother called Mary? And are not his brothers James and Joseph and Simon and Judas? And are not all his sisters with us? Where then did this man get all these things?" And they took offense at him.'(Matthew 13:54-57).
The danger of 'See-finish-ism' is that it blinds us to the treasure that may lie deep within the person involved. We may think we've seen it all, but there may be so much more. Your pastor may still have a Word in him that can turn your life around, even though he's had struggles in his family. Your spouse is God's gift to you, and his/her prayers over your life can alter your destiny; a word of advice from them can change your fortunes. Even your children-little as they are, yeah you cleaned their messy diapers! – may have something to say or teach you that could be a major blessing.
God can use anybody, remember that!
Don't write people off just because you know their story! Jesus' townsmen did, and what is the conclusion of the matter? They lost out!
My dearest Dr. Luch Luch, inspiring my life since 1660.
"See-finishism"…….embued with so much wisdom.
True words. I'm making amends.
Thank you so much for your feedback. I am making amends too.
This is very common, worse still in the house of God and the body of Christ in general.
Nice piece Doc, keep the flag flying.
Thank you so much Wari, I appreciate the feedback.


text:
Mental health support, not increased policing, needed during pandemic, study finds
by Simon Fraser University
A new study finds that while most crime types declined across Canada and internationally as a result of the COVID-19 social restrictions, mental health-related incidents remained relatively stable, counter to claims that mental health related incidents increased across the nation as a result of the pandemic related restrictions.
The study, published in the Canadian Journal of Criminology and Criminal Justice, suggests that calls for additional policing services to respond to mental-health related demands may be unwarranted and that focus on preventative measures is key.
"Our findings suggests that police resourcing may not be an important focus during an exceptional event such as a pandemic," says Simon Fraser University criminology professor Martin Andresen, the study's co-lead. "Rather, we suggest that funding may be better spent on preventative strategies that improve evidence-based predictors of mental health such as housing, basic income, and social services."
"For instance, research has shown that the impact of the pandemic was not distributed evenly and disproportionally impacted women, people experiencing illness, people with previous mental health-related issues, and other marginalized groups," says Andresen.
Andresen and Wilfrid Laurier University assistant professor Tarah Hodgkinson analyzed mental health-related calls from 13 police jurisdictions across Canada from March 2019 to November 2021.
They used open-source data from the Canadian Center for Justice and Community Safety Statistics, Statistics Canada to explore the impact of the COVID-19 pandemic on police-reported mental health-related calls.
Digging into the data
Researchers looked at calls for property and violent crimes, which decreased in almost all jurisdictions. One notable exception was London, Ontario where violent crime increased by 30 percent during the pandemic.
The team found that Mental Health Act apprehensions increased in 10 out of 13 police services with the exception of Calgary and Regina. Apprehensions are made by police officers when individuals are considered a danger to themselves or others, and result in a medical assessment at a medical facility.
Ontario Provincial Police, Royal Newfoundland Constabulary, Toronto, and Waterloo Region all recorded statistically significant increases in Mental Health Act (MHA) apprehensions (47, 12, 10, and 13 percent respectively). Meanwhile researchers found an almost complete lack of MHA apprehensions in Vancouver reported to Statistics Canada.
They generally found no increases in calls to police for suicide or suicide attempts during COVID-19 with statistically significant decreases in Halton Region, Toronto, Vancouver and York Region.
However, calls to police categorized as mental health (other) increased in Toronto, Regina, and Ottawa. In particular, researchers found a large-magnitude increase in volume for Toronto.
A rise in mental health (other) calls to police could suggest that certain jurisdictions may be more vulnerable and in need of improved or expanded social services.
Instead of dedicating additional funding to police services during a crisis, researchers suggest income supports and support for those struggling with addictions in vulnerable and marginalized communities will help address the mental health issues reducing the need for police assistance.
More information: Martin Andresen et al, Stumbling from One Disaster to Another: The COVID-19 Pandemic and Mental Health Calls for Police Service across Canada, Canadian Journal of Criminology and Criminal Justice (2022). DOI: 10.3138/cjccj.2022-0016
Provided by Simon Fraser University
Citation: Mental health support, not increased policing, needed during pandemic, study finds (2022, October 11) retrieved 31 January 2023 from https://phys.org/news/2022-10-mental-health-policing-pandemic.html
Researchers study COVID-19 and its impact on crime in Vancouver, Canada
Study shows that eggshells of large, flightless birds evolved along different tracks
'Snapshots' of translation could help us investigate cellular proteins
Defunding the police: Calls not easily transferable to other sectors without significant expenditures, adjustments
Why some doctors want to defund the police
Better outcomes when police, ambulance and mental health services attend 111 mental health emergencies together
How do encounters with police affect the mental health of black Americans?
Study: In Tempe, AZ, COVID-19 pandemic had stronger impact on policing and crime than George Floyd's death
Schoolkids lost third of a year's learning to pandemic: study
Stereotyping veterans as heroes may limit their future careers
Preserving endangered languages as 3D shapes
Urban elites seize most of the benefits of big cities, finds study
Rural water infrastructure development research demonstrates ethnically, racially uneven economic outcomes


My goal is to produce a print that pleases me. The contributing factors to such a print are story, composition, and tonal quality. To find such pictures I depend upon luck and intuition.
I use small format (35mm) and medium format cameras and work with film, chemicals, and silver gelatin fiber paper.


Once again as we approach November, Diabetes Awareness Month, the Gaylord Lions Club along with the American Diabetes Association, encourages diabetics to have a dilated eye exam.
With 16 million diabetics in our nation today, over 25 percent will develop some form of diabetes-related complications, whether diabetic retinopathy (damage to the blood vessels in the retina), cataracts (the clouding of the eye's lens) or glaucoma (an increase of fluid pressure inside the eye causing damage to the optic nerve). Serious complications can be prevented with early diagnosis.
The Lions Club International was founded in 1919 as a social club until 1925 when Helen Keller appeared at their meeting. She challenged the Lions to be "knights of the blind in the crusade against darkness." From this challenge, the Lions established the white cane as a symbol of independence for blind people; founded some of the first leader-dog schools for the blind; and established the majority of the world eye banks. That's not to mention the number of local vision, hearing, and diabetes cases handled through the club.
If you are interested in learning more about the projects of the Gaylord Lions Club, or joining our club we welcome your questions.
Please contact Tom Otto at 732-7840, or Madge Smith, 732-7250.


Microsoft has revamped SQL Server Management Studio to run in the Visual Studio 2010 shell. It has also introduced a new approach to database development called SQL Server Data Tools. The thinking here is to focus on the code that defines the database schema. Using the Data Tools, you can create a database project, import or build the schema, and then test it using a local database instance. Since the schema is text-based, it can be checked into version control. There are also tools to compare and refactor schemas, with warnings where inconsistencies may be introduced.


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Coach says airline agent asked her to 'prove' biracial baby was her son
Southwest Airlines apologized Tuesday to the University of California Berkeley women's basketball coach Lindsay Gottlieb after she claimed an airline employee stopped her from boarding because the worker didn't believe her 1-year-old biracial son was hers.A desk agent questioned Gottlieb ahead of a flight from Denver to Oakland Sunday, saying she "had to 'prove' that he was my son, despite having his passport," Gottlieb said in a series of tweets."She said because we have different last name. My guess is because he has a different skin color," Gottlieb tweeted Monday. Gottlieb was traveling with her fiance, Patrick Martin, the boy's father, who is black.Airlines aren't required to match the last name of a child and guardian for domestic flights."We have reached out to Ms. Gottlieb directly to address her concerns and will utilize the situation as a coaching opportunity for our employee," Southwest said in a statement. "We apologize if our interaction made this family uncomfortable — that is never our intention."Gottlieb said Tuesday the encounter was hurtful, but she appreciates Southwest's apology."I felt that in this situation it was my responsibility to say 'Hey, this isn't OK,'" Gottlieb said in a statement to The Associated Press. "I hope the coverage this has received can serve as a learning opportunity and that all families — regardless of how 'traditional' they may or may not look — are treated with dignity and respect."Gottlieb, Martin and their child were eventually allowed to board.
Southwest Airlines apologized Tuesday to the University of California Berkeley women's basketball coach Lindsay Gottlieb after she claimed an airline employee stopped her from boarding because the worker didn't believe her 1-year-old biracial son was hers.
A desk agent questioned Gottlieb ahead of a flight from Denver to Oakland Sunday, saying she "had to 'prove' that he was my son, despite having his passport," Gottlieb said in a series of tweets.
"She said because we have different last name. My guess is because he has a different skin color," Gottlieb tweeted Monday. Gottlieb was traveling with her fiance, Patrick Martin, the boy's father, who is black.
Airlines aren't required to match the last name of a child and guardian for domestic flights.
"We have reached out to Ms. Gottlieb directly to address her concerns and will utilize the situation as a coaching opportunity for our employee," Southwest said in a statement. "We apologize if our interaction made this family uncomfortable — that is never our intention."
Gottlieb said Tuesday the encounter was hurtful, but she appreciates Southwest's apology.
"I felt that in this situation it was my responsibility to say 'Hey, this isn't OK,'" Gottlieb said in a statement to The Associated Press. "I hope the coverage this has received can serve as a learning opportunity and that all families — regardless of how 'traditional' they may or may not look — are treated with dignity and respect."
Gottlieb, Martin and their child were eventually allowed to board.


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Redseed Game Studio
Redseed Game Studio Blog where you can read all about video games and how we develop them.
Development of Niara: Rebellion Of the King
Right now at Redseed we are working on our new video game project called Niara: Rebellion Of the King. This game will be a visual novel RPG and will be released on Google Play Store in April 2019.
The progress in the development of the game has been good. We have completed the development of the dialogues, the quest system, inventory system, map system and the battle system. We need to start working on the shop system and then start creating the content for the game. This content will include the story which the game will follow as well as the different quests which the player will need to finish. We are developing this game in Renpy.
Here are some screenshots of the game:
How to learn...programming languages? Or really any hands on skill.
Not by reading its book. No.
If you are learning a new programming language or framework (is that correct?) you should be doing step by step tutorials which focus on building a mini app or project. Like, a good starting point would be to make a 'hello world' app. Do not try to read the tomes which the scientists and scholars recommend you. That works for scientists and scholars. If you are an engineer or developer then you just cannot afford to waste weeks in reading volume after volume of books to learn a programming language. You need to learn the next best thing as quickly as possible and deliver quality software in time.
That said, books are good for references and of course if you are trying to build a new programming language then you need to do a lot of research but hey, as I said, thick tomes are good for researchers not for developers.
Niara: ROTK Devlog | What type of video game should you make?
The whole point of us making video games was to portray stories to the people in an engaging medium. Video games are one of the most engaging mediums using which one can portray a story very effectively.
However, there are video game genres which are not very suitable for telling stories. These can be casual games or puzzle games. We really avoided these genres from the get go because we knew portraying a deep and intricate story using a puzzle game was next to impossible. But then again, there maybe exceptions out there.
We decided that we needed to make this game an RPG. RPG (Role Playing Games) are one of the most popular game genres out there and they have been used by several studios across the world to portray very deep and intricate stories. Some of the very famous examples being The Elder Scrolls and the Witcher series.
However, we are a very very small game studio. So, no way we would have been able to pull off a Witcher level video game. We knew our limits and therefore we de…
Niara: ROTK Devlog | How to choose the right game engine?
When you are trying to make a video game then you must almost always go with a game engine. Sure there are graphic libraries for most of the popular programming languages in the world but making a game which will sell well in today's day and age using just a graphics library is nearly impossible. Python has pygame library, C has SDL but making a game using just these libraries is a nightmare. We must always go for an engine. Game engines make our lives easier and we are able to build the games faster.
Now, before you choose a game engine for your game you have to decide what type of game you want to make. This we have discussed in our previous article which you can read here: Niara: ROTK Devlog | What type of video game should you make?
We decided to make a visual novel role playing game in Renpy. And we decided to go with 2D graphics. This was because we are a very small indie game studio. If you are also a small indie game studio or you are just starting out then you should also …
Copyright (c) 2018 Ayush Raj. All rights reserved
niararotk
visualnovelrpg


THE GIRL AND THE FISH.
THERE was once a girl who used to go to the river to fetch water, but when she went she was never in a hurry to come back, but stayed so long, that they made up their minds to watch her. So one day they followed her to the river, and found when she got there she said something (the reciter forgets the words), and a fish came up and talked to her; and she did not like to leave it, for it was her sweetheart. So they went next day to the river to see if the fish would come up, for they remembered what the girl said, and used the same words. Then up came the fish immediately, and they caught it, and took it home, and cooked it for dinner,—and a part they set by, and gave it to the girl when she came in. Whilst she was eating, a voice said, "Do you know what you are eating? I am he you have so often talked with. If you look in the pig's tub you will see my heart." Then the voice told her to take the heart, and wrap it up in a handkerchief, and carry it to the river. When she got to the river she would see three stones in the water; she was to stand on the middle stone and dip the handkerchief three times into the water. All this she did, and then she sank suddenly, and was carried down to a beautiful place, where she found her lover changed from a fish into his proper form, and there she lived happily with him for ever. And this is the reason why there are mermaids in the water.


Work has been crazy busy leading up to last weekend, so I ended up driving the car more than I would have ideally liked. Sometimes needs must and this week I was able to return to commuting on the bike and the last two days have been polar opposites in terms of weather and riding achieved.
On Tuesday I had to return a key to the local council and took the Ducati into town for the job. The sun was up, the wind was down and it was a lovely, lovely day. After dropping the key off and grabbing a coffee, I planned my route back to work. The direct route was primarily dual carriageways, which didn't appeal, so in what can only be described as 'stealing a ride' I took the much more desirable scenic route.
Riding out in the mid morning sunshine, the roads were quiet and the Ducati and I had a field day, but I can't say the same thing about some of the bugs we met en-route. One of which can be best described as 'full'.
After the joy of yesterday, today was windy and rainy. Not being afraid of the rain, the Ducati and I headed back out and the Rosso tyres she's sporting are fantastic in the wet. I was maintaining a good speed and making good progress despite the damp conditions and both wheels dealt with any surface water brilliantly. I didn't have a single moment where I feared I would loose grip. That's confidence building right there.
My mate Dean and I are all booked in for a trackday up at Rockingham with Bike Magazine in August. Dean also has a black 1098s so it should be a good laugh. The only downside is Rockingham is at least three hours from home. Looks like we'll be sharing the driving.


Head Coach Dermot Drummy has today confirmed that two more players, Shamir Fenelon and Gavin Tomlin, are being released when their contracts expire on June 30.
Dermot said: "Both Gavin and Shamir have been excellent pros since I came to the club.
Tomlin made 57 appearances including 43 starts after joining the club in 2014. Fenelon, who joined Reds in 2015, made 32 appearances including 10 starts.


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Goodyera repens (L.) R.Br., 1813è una piccola pianta erbacea perenne dai delicati fiori bianchi, appartenente alla famiglia delle Orchidacee.

Etimologia
Il nome generico (Goodyera) è stato creato dal botanico scozzese Robert Brown (1773 – 1858) in onore di un altro botanico: John Goodyer (1592-1664), un naturalista inglese considerato un precursore della botanica moderna.

Il termine specifico (repens) significa "strisciante" e fa riferimento al tipo di portamento della pianta.

In lingua tedesca questa pianta si chiama Moosorchis oppure Netzblatt oppure Kriechstendel; in francese si chiama Goodyère rampante; in lingua inglese si chiama Creeping Lady's-tresses.

Descrizione

È una pianta erbacea perenne alta normalmente da 5 a 30 cm. La forma biologica di questa orchidea è geofita rizomatosa (G rizh), ossia è una piante con un particolare fusto sotterraneo, detto rizoma, che ogni anno si rigenera con nuove radici e fusti avventizi. È un'orchidea terrestre in quanto contrariamente ad altre specie, non è "epifita", ossia non vive a spese di altri vegetali di maggiori proporzioni (ha cioè un proprio rizoma).

Radici
Le radici sono secondarie da rizoma (vengono emesse dai nodi del rizoma).

Fusto
Parte ipogea: la parte sotterranea consiste in un lungo rizoma gracile, stolonifero e radicante ai nodi. È strisciante e di colore bianco-roseo. Tende a ricoprire un'area più ampia di quella che farebbe supporre in base alle dimensioni visibili della pianta.
Parte epigea: i fusti aerei sono ascendenti, delicati e ramificati; spesso a portamento prostrato o rampante sul terreno. Sono pubescenti nella parte alta.

Foglie

Foglie basali: è una pianta dotata di diverse foglie (da 4 a 7) radicali o basali disposte a rosetta, addensate e a forma intera, ovato-bislunga (quasi spatolata) con apice appuntito; possiedono un picciolo allargato. La lamina è verde ed è solcata da diverse nervature biancastre e parallele ma anche reticolate (5 nervi anastomosati – per la flora spontanea delle orchidee italiane questa è l'unica eccezione), mentre la consistenza è carnosa; la superficie è glabra. Il portamento è patente. Dimensioni del picciolo: larghezza 2 – 3 mm; lunghezza quanto la lamina della foglia. Dimensioni della foglia: larghezza 8 – 12 mm; lunghezza 18 – 25 mm.
Foglie cauline: quelle del fusto (2 – 3 foglie cauline) sono ridotte (bratteiformi), a forma lanceolato-lineare e sono amplessicauli (abbracciano il fusto per tutta la loro lunghezza) tipo guaina. Lunghezza delle foglie cauline: 5 – 7 cm.

Infiorescenza

L'infiorescenza è sciolta di tipo spiciforme. La disposizione dei fiori (da 7 a 36) è più o meno spiralata e unilaterale. Tutta l'infiorescenza è pubescente-glandulosa. Alla base del ovario (sotto il perigonio) sono presenti delle brattee erbacee a forma lanceolata; queste sono più lunghe dell'ovario. L'infiorescenza è inoltre un po' incurvata all'apice. Lunghezza dell'infiorescenza: 5 – 15 cm.

Fiore

I fiori sono ermafroditi ed irregolarmente zigomorfi, pentaciclici (perigonio a 2 verticilli di tepali, 2 verticilli di stami (di cui uno atrofizzato), 1 verticillo dello stilo). Dimensione dei fiori: 3 – 7 mm.

Formula fiorale: per queste piante viene indicata la seguente formula fiorale:
P 3+3, [A 1, G (3)]

Perigonio: il perigonio è composto da 2 verticilli con 3 tepali ciascuno (3 interni e 3 esterni) di forma più o meno lanceolato-ovata, più o meno liberi e patenti. Il primo verticillo (esterno) ha 3 tepali conniventi di colore biancastro e sono vistosamente pubescenti-glandulosi, con apice acuto e margini lievemente ripiegati all'interno; il loro portamento ricorda quello di una campanula e sono quasi ad angolo retto rispetto all'ovario. Nel secondo verticillo (interno) il tepalo centrale (chiamato "labello") è molto diverso rispetto agli altri due laterali che si presentano più ottusi e più simili ai tre tepali esterni. Sono inoltre aderenti ai tepali esterni. Lunghezza dei tepali: 3 - 4 mm.
Labello: il labello (semplice – non formato da due parti distinte) è il tepalo centrale più interno a forma ovato-gozzuta (ossia si presenta "saccato" - a forma di sacco) e termina con una "lingua" (ovato-triangolare) ripiegata verso il basso con una insenatura centrale disposta longitudinalmente. Al suo interno si trovano gli organi di riproduzione (il ginostemio). Il labello non è speronato come in altri generi della stessa famiglia. Il labello è nettarifero. Dimensione del labello: larghezza 1,4 – 3,2 mm; lunghezza 1,8 – 4,8 mm. Lunghezza della linguetta: 4 mm.
Ginostemio: lo stame con le rispettive antere (si tratta di una antera biloculare) è concresciuto con lo stilo e forma una specie di organo colonnare chiamato ginostemio. In particolare nelle specie di questo genere le antere sono localizzate sul dorso del ginostemio e il polline è a consistenza granulare. Il colore del ginostemio è giallo alla base e rosso all'apice. L'ovario, infero e sessile, è formato da tre carpelli fusi insieme; inoltre è lievemente pubescente.
Fioritura: da giugno ad agosto.

Frutti

Il frutto è una capsula più o meno ovoidale con alcune coste. È deiscente lungo tre di queste coste. Al suo interno sono contenuti numerosi minutissimi semi (sono probabilmente i semi più piccoli della famiglia delle Orchidee). Questi semi sono privi di endosperma e gli embrioni contenuti in essi sono poco differenziati in quanto formati da poche cellule. Queste piante vivono in stretta simbiosi con micorrize endotrofiche, questo significa che i semi possono svilupparsi solamente dopo essere infettati dalle spore di funghi micorrizici (infestazione di ife fungine del genere Rhizoctonia). Questo meccanismo è necessario in quanto i semi da soli hanno poche sostanze di riserva per una germinazione in proprio..

Biologia
La riproduzione di questa pianta avviene in due modi:
per via sessuata grazie all'impollinazione degli insetti pronubi (Impollinazione entomofila).
per via vegetativa in quanto i rizomi possono emettere gemme avventizie capaci di generare nuovi individui.

Distribuzione e habitat
Geoelemento: il tipo corologico (area di origine) è Circumboreale - Eurosiberiano, ma anche Nord Americano.
Diffusione: in Italia è considerata rara e si trova nelle Alpi (ad esclusione delle province di Novara, Varese e Como) e negli Appennini settentrionali e centrali (negli Appennini è probabilmente avventizia). Fuori dall'Italia è presente nell'Europa occidentale (sui rilievi non è segnalata nelle Alpi Dinariche) e comunque è presente fino ai Carpazi (è l'orchidea più diffusa in Scandinavia). È spontanea anche nelle zone montuose dell'America del nord. In Italia è da considerare una "pianta relitto", ossia probabilmente derivata dai climi freddi e asciutti dei periodi postglaciali
Habitat: l'habitat tipico sono le stazioni alpine ombrose e umide (pinete montane a Pino nero e Pino silvestre). È una pianta che facilmente si trova a contatto con i cuscinetti di muschio nei sottoboschi di aghifoglie. Sviluppa quindi una simbiosi micorrizica con i microrganismi di questo ambiente. Il substrato preferito è sia calcareo che siliceo con pH neutro e bassi valori nutrizionali del terreno che deve essere secco.
Diffusione altitudinale: sui rilievi queste piante si possono trovare dai 400 fino a 2000 ; frequentano quindi i seguenti piani vegetazionali: montano e subalpino.

Fitosociologia
Dal punto di vista fitosociologico la specie di questa voce appartiene alla seguente comunità vegetale:
Formazione: delle comunità forestali
Classe: Vaccinio-Piceetea excelsae
Ordine: Picetalia excelsae

Sistematica
Queste orchidee sono assegnate alla tribù delle Cranichideae caratterizzate da antere persistenti dotate di polline granulare.

Il numero cromosomico di G. repens è: 2n = 30.

Variabilità
I fiori delle varietà americane hanno colori più vicini al verde (verdognolo) oppure al bianco crema e il colore delle foglie è più scuro (verde-blu-oliva). In quest'area si sono individuate due varietà (queste due varietà non sono presenti in Europa e vengono considerate sinonimi della specie nominale):
Goodyera repens var. repens: individui a foglie verdi e semplici (lamina poco reticolata).
Goodyera repens var. ophioides Fernald: le venature della foglie sono circondate da ampie fasce biancastre (lamina molto reticolata).

Sinonimi
La specie Goodyera repens ha avuto nel tempo diverse nomenclature. L'elenco che segue indica alcuni tra i sinonimi più frequenti:
Satyrium repens L. (1753) (basionimo)
Epipactis repens (L.) Crantz (1769)
Serapias repens (L.) Vill. (1787)
Neottia repens (L.) Sw. (1800)
Orchis repens (L.) Eyster ex Poir. (1805)

Usi

Farmacia
In alcune zone questa pianta trova qualche impiego nella medicina popolare: una infusione delle foglie contro il mal di denti o per migliorare l'appetito; le radici a volte vengono usate nel trattamento di problemi vescicali.

Conservazione
Come tutte le orchidee è una specie protetta e quindi ne è vietata la raccolta e il commercio ai sensi della Convenzione sul commercio internazionale delle specie minacciate di estinzione (CITES).

Galleria d'immagini

Note

Bibliografia

Voci correlate
Orchidee selvatiche in Italia

Altri progetti

Collegamenti esterni

Goodyera repens Flora delle Alpi Marittime Database
Goodyera repens eFloras Database
Goodyera repens Flora Europaea (Royal Botanic Garden Edinburgh) Database
Goodyera repens Flora Italiana - Schede di botanica
Goodyera repens G.I.R.O.S. - Gruppo Italiano per la Ricerca sulle Orchidee Spontanee - Database
 GRIN Database
Goodyera repens IPNI Database
Goodyera repens Royal Botanic Gardens KEW - Database
Goodyera repens Tropicos Database
Goodyera repens Catalogazione floristica - Università di Udine
 ZipcodeZoo Database

Cranichideae
Taxa classificati da Linneo
Taxa classificati da Robert Brown


Flexible Coupling are used to transmit torque from one shaft to another when the two shafts are slightly misaligned.
Traditionally, gear couplings were used in these applications but required constant lubrication to extend their life.
Such machines are usually coupled by a Heavy Duty SWC-BF Type Cardan Shaft where more than 4-6 degrees of angle exists between the cardan coupling and the machines.
WHAT IS A RIGID COUPLING?
Rigid Coupling serve as components to time, join, or align shafts at lower speeds and torque, especially when zero backlash is desired.
What Do You Know About Heavy Duty Cardan Shafts?
The universal coupling such as Heavy Duty Cardan Shaft makes use of the characteristics of its mechanism so that the two shafts are not on the same axis, and the two shafts connected can be rotated continuously when there is an Angle between axes, and the torque and motion can be transmitted reliably.


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Q: Change folder name with the name between ( ) with powershell I have a folder structure and in all the folders there is a part between ( ).
Like "folder example(12345)"
Now I must rename all the folders with the name that is between ( ).
So the result is folder "12345"
I have an beginning and then I am stuck.
# Root path
$RootPath = "E:\Test\"
# Rename folders
Get-ChildItem -Path $RootPath -Filter '*()*' -Directory -Depth 0 | Rename-Item -NewName

A: This command will rename folder example(12345) to 12345:
# Root path
$RootPath = "E:\Test\"
# Rename folders
Get-ChildItem -Path $RootPath -Filter '*(*)*' -Directory -Depth 0 | Rename-Item -NewName { $_.Name -replace '.*\((.+?)\).*', '$1' }

What the rename command does:
Get-ChildItem            Get items in the folder
    -Path $RootPath      specified in this variable.
    -Filter '*(*)*'      Only take items that match this search
    -Directory           Only take directories
    -Depth 0             Only look in the current directory level
    |                    Forward the result to the next command
    Rename-Item          Rename the incoming items
    -NewName             Specify what the new name should be
    {
        $_.Name          Take the name of the item to be renamed
        -replace         replace that name
        '.*\((.+?)\).*', matching a regular expression
        '$1'             with the first capture group (see below for more explanation)
    }

The regex can be further explained:
.*    Match any character 0 or more times
\(    Match a literal left parenthesis
(.+?) Capture a group containing any character occurring more than once, but as few times as possible.
\)    Match a literal right parenthesis
.*    Match any character 0 or more times

The captured group can be reused by referring to it with $1.
A demonstration:

A: Based on the answer of Saaru Lindestøkke I can confirm that the script should be working fine.
Here is the code that worked for me:
# Root path
$RootPath = "C:\Temp\"
# Rename folders
Get-ChildItem -Path $RootPath -Filter '*(*)*' | Rename-Item -NewName { $_.Name -replace '.*\((.+?)\).*', '$1' }

I took your example, changed
*()*

to
*(*)*

otherwise it doesn't find anything since it looks for Wildcard()Wildcard and I also changed $RootPath to "C:\TEMP" (It works both with and without \ after TEMP) and only ran the part of Get-ChildItem to see the result. It would always find the folder(s) I created.
I removed as well "-Directory -Depth 0" since I didn't know if it was needed but never the less it should be working.


Today I would like to unveil my latest PDF pattern, an enhanced version of my Load-it-Up Laptop Slipcase, originally offered a few weeks back as a free tutorial.
The revised design includes a grab handle, an interior zippered pocket and an optional patch pocket underneath the bellows pocket.
This means that not only will this case still carry around your laptop, charge cord and mouse, the additional features in this version will also protect your important small stuff (like USB sticks) and — if you want — even a pen or stylus and notepad.
Plus, with a handle, it's now much easier to grab and go.
In addition, as a bonus for those of you who avoid doing math as much as possible, I put together a couple of charts of common laptop measurements (in both metric and imperial) so that you can simply look up the values and then cut your fabric to suit.
Here are some pics of my test project.
The laptop for which this case was made is a netbook, meaning that its screen size is in the neighbourhood of 10". It fits inside while "wearing" a folio style case, which is why it's bigger than the first one that I made for a similar size computer.
Here is the new interior zippered pocket that can safely house your memory cards and USB sticks. It's sized to do just that job (meaning that it's not deep) but of course, you can use a larger piece of pocket lining fabric to make it bigger if that's your preference.
The grab handle is padded for carrying comfort.
For those who like to know, the fabric above is Betz White's Stitch from a collection that she did for Robert Kaufman (part of a gift package that I received two years ago).
Naomi volunteered to test this pattern for a larger laptop and here are a few photos of her results.
This is definitely a version for someone who wants to stand out, what with the hot pink lining and pink floral print!
FYI, Naomi's laptop is 15" wide by 1" thick by 10" deep, which is much bigger than either of mine.
And here is a look at the bonus pen and notebook (or whatever) pocket under the bellows pocket.
The Load-it-Up Laptop Slipcase is a quick sew and easy to put together, even with the additional features. You can definitely whip this up in a leisurely day.
The PDF is 23 pages long with 45+ photos to assist you. Happy sewing!
you can purchase from my Makerist Shop.
NOTE: Due to the nature of digital products, all PDF sales are final and non-refundable.
That's a nice looking laptop case. Happy pattern sales!


When reading about multiple myeloma – and more particularly about research – you're likely to come across terms like "complete response" (CR) or "partial response" (PR).
Please be aware that different studies may apply different definitions to the same terms.
Below are the acronyms of the response categories used by the International Myeloma Working Group.
For a definition of these response categories, please consult page 26 of the Multiple Myeloma Patient Handbook.
Remission is the complete or partial disappearance of the signs and symptoms of myeloma.
Relapse is when signs and symptoms of myeloma return after a period of improvement.
Refractory is the term used to describe myeloma that does not respond to treatment.
Many patients can live full and productive lives for years after diagnosis, even after experiencing a relapse. Watch the video below and to hear stories from other patients who have experienced a relapse. For more information on treatment options, click here.