Testing and Evaluating Dye Color: A Study Using Spectrophotometry on Technicolor and Eastman Color Motion Picture Film

The history of color motion picture film is linked to two companies: Technicolor and Eastman Kodak. Technicolor was formed in 1915 by Herbert Kalmus, Daniel Comstock, and W. Burton Wescott, who realized that color film could enhance motion picture entertainment. Introduced in 1932, it was a three-color process that was widely used into the late 1950s. The three-color process utilized a camera with one lens that exposed three black and white negatives simultaneously by splitting the incoming visible light through a prism. The prism would create two beams of light which would then expose one film negative to green light and the rest of the split light exposed a red and blue bi-pack comprised of two film negatives placed emulsion to emulsion.1 One film negative would be sensitized to blue light while the other to red light, once exposed, the two negatives would be separated to be processed. The Eastman Color process, developed by Eastman Kodak Company in 1950, was similar to Technicolor. Eastman Color used three light-sensitive emulsions that were sensitized to red, green, and blue light which were then coated onto a single film base.2 Having all three emulsion layers on one film support does away with the bulky three-color process camera used in the Technicolor process. Eastman Color prints were less costly to produce than Technicolor prints. This was because Eastman Color only used one film negative versus Technicolor�s three. Within a decade, Eastman Color films began to show evidence of fading in the negatives and prints. The film industry�s shift from Technicolor to Eastman Color film resulted in a shift in the color, tone, and color balance of the film prints. Technicolor three-color dye process produces a very stable color 1 Scott Higgins, Demonstrating Three-Colour Technicolor: Early Three-Colour Aesthetics and Design, Film History 12, 4 (2000): 358-383. 2 Doug Nishimura, Email correspondence with author, June 3, 2015. 2 that doesn�t easily degrade with hues that were “warm,” whereas Eastman Color tended to be “cooler” and almost neon-like. In this thesis, I will discuss the differences in dyes used by Technicolor and Kodak and illustrate how the dyes have altered over time. To do so, I have made measurements on dyed film test strips by using a spectrophotometer that was created specifically for this testing. I sampled twenty-five test dyed film strips and twenty-five Technicolor film reels. In addition, a single Eastman Color film reel was tested. This sample size of fifty-one items yielded raw data that was calculated into absorbance and transmittance wavelengths. These spectral curves allow for comparison between the dyes used and for an evaluation of the colors. In the discussion, I describe why knowledge of dye fading is important to museums and how spectral information on dyes could improve film preservation efforts. This data is analyzed by reviewing scholarly journals, case studies, and first-hand accounts of dye tests in an effort to further the knowledge for museums and professionals with a focus on information on motion picture film, preservation, and spectrophotometry

Bedtime Salivary Cortisol as a Screening Test for Cushing Syndrome in Children

Background Diagnosing Cushing syndrome (CS) can be challenging. The 24-hour urine free cortisol (UFC) measurement is considered gold standard. This is a laborious test, dependent on correct urine collection. Late-night salivary cortisol is easier and is used as a screening test for CS in adults, but has not been validated for use in children. Objective To define liquid chromatography tandem mass spectrometry (LC-MS/MS)-based cutoff values for bedtime and morning salivary cortisol and cortisone in children, and validate the results in children with and without CS. Methods Bedtime and morning salivary samples were collected from 320 healthy children aged 4 to 16 years. Fifty-four patients from the children’s outpatient obesity clinic and 3 children with pituitary CS were used for validation. Steroid hormones were assayed by LC-MS/MS. Cutoff levels for bedtime salivary cortisol and cortisone were defined by the 97.5% percentile in healthy subjects. Results Bedtime cutoff levels for cortisol and cortisone were 2.4 and 12.0 nmol/L, respectively. Applying these cutoff levels on the verification cohort, 1 child from the obesity clinic had bedtime salivary cortisol exceeding the defined cutoff level, but normal salivary cortisone. All 3 children with pituitary CS had salivary cortisol and cortisone far above the defined bedtime cutoff levels. Healthy subjects showed a significant decrease in salivary cortisol from early morning to bedtime. Conclusions We propose that bedtime salivary cortisol measured by LC-MS/MS with a diagnostic threshold above 2.4 nmol/L can be applied as a screening test for CS in children. Age- and gender-specific cutoff levels are not needed.publishedVersio

Analgesic effect of recombinant GABAergic precursors releasing omega-conotoxin MVIIA in a model of peripheral nerve injury in rats

Development of chronic pain has been attributed to dysfunctional GABA signaling in the spinal cord. Direct pharmacological interventions on GABA signaling are usually not very efficient and often accompanied by side effects due to the widespread distribution of GABA receptors in CNS. Transplantation of GABAergic neuronal cells may restore the inhibitory potential in the spinal cord. Grafted cells may also release additional analgesic peptides by means of genetic engineering to further enhance the benefits of this approach. Conopeptides are ideal candidates for recombinant expression using cell-based strategies. The omega-conopeptide MVIIA is in clinical use for severe pain marketed as FDA approved Prialt in the form of intrathecal injections. The goal of this study was to develop transplantable recombinant GABAergic cells releasing conopeptide MVIIA and to evaluate the analgesic effect of the grafts in a model of peripheral nerve injury-induced pain. We have engineered and characterized the GABAergic progenitors expressing MVIIA. Recombinant and nonrecombinant cells were intraspinally injected into animals after the nerve injury. Animals were tested weekly up to 12 weeks for the presence of hypersensitivity, followed by histochemical and biochemical analysis of the tissue. We observed beneficial effects of the grafted cells in reducing hypersensitivity in all grafted animals, especially potent in the recombinant group. The level of pain-related cytokines was reduced in the grafted animals and correlation between these pain markers and actual behavior was indicated. This study demonstrated the feasibility of recombinant cell transplantation in the management of chronic pain

Chapter 19 - Cell transplantation for reducing neuropathic pain after SCI

The transplantation of cells into the CNS has been considered a promising long-term therapeutic approach in the management of neurological disorders. Cell transplantation can provide replacement of lost or damaged neurons and supportive cells as well as a sustained source of pharmacologically active beneficial molecules. Both of these approaches, and their combination, have been explored for the management of chronic pain over the past several decades. Neuropathic pain following SCI is a challenging clinical target, which may be particularly amenable to the direct interventive strategies provided by cell transplantation. This chapter reviews cell transplantation approaches that have been considered for chronic pain amelioration, both as a source of analgesic substances and as disease modification, in preclinical models and early clinical studies. Recent combination strategies such as ex vivo gene therapy enhancement and inclusion of physical exercise may maximize long-term beneficial potential in the management of SCI neuropathic pain

Development of a Phantom Limb Pain Model in Rats: Behavioral and Histochemical Evaluation

Therapeutic strategies targeting phantom limb pain (PLP) provide inadequate pain relief; therefore, a robust and clinically relevant animal model is necessary. Animal models of PLP are based on a deafferentation injury followed by autotomy behavior. Clinical studies have shown that the presence of pre-amputation pain increases the risk of developing PLP. In the current study, we used Sprague-Dawley male rats with formalin injections or constriction nerve injury at different sites or time points prior to axotomy to mimic clinical scenarios of pre-amputation inflammatory and neuropathic pain. Animals were scored daily for PLP autotomy behaviors, and several pain-related biomarkers were evaluated to discover possible underlying pathological changes. Majority displayed some degree of autotomy behavior following axotomy. Injury prior to axotomy led to more severe PLP behavior compared to animals without preceding injury. Autotomy behaviors were more directed toward the pretreatment insult origin, suggestive of pain memory. Increased levels of IL-1β in cerebrospinal fluid and enhanced microglial responses and the expression of NaV1.7 were observed in animals displaying more severe PLP outcomes. Decreased expression of GAD65/67 was consistent with greater PLP behavior. This study provides a preclinical basis for future understanding and treatment development in the management of PLP

Regulation of defective mitochondrial DNA accumulation and transmission in C. elegans by the programmed cell death and aging pathways.

The heteroplasmic state of eukaryotic cells allows for cryptic accumulation of defective mitochondrial genomes (mtDNA). Purifying selection mechanisms operate to remove such dysfunctional mtDNAs. We found that activators of programmed cell death (PCD), including the CED-3 and CSP-1 caspases, the BH3-only protein CED-13, and PCD corpse engulfment factors, are required in C. elegans to attenuate germline abundance of a 3.1-kb mtDNA deletion mutation, uaDf5, which is normally stably maintained in heteroplasmy with wildtype mtDNA. In contrast, removal of CED-4/Apaf1 or a mutation in the CED-4-interacting prodomain of CED-3, do not increase accumulation of the defective mtDNA, suggesting induction of a non-canonical germline PCD mechanism or non-apoptotic action of the CED-13/caspase axis. We also found that the abundance of germline mtDNAuaDf5 reproducibly increases with age of the mothers. This effect is transmitted to the offspring of mothers, with only partial intergenerational removal of the defective mtDNA. In mutants with elevated mtDNAuaDf5 levels, this removal is enhanced in older mothers, suggesting an age-dependent mechanism of mtDNA quality control. Indeed, we found that both steady-state and age-dependent accumulation rates of uaDf5 are markedly decreased in long-lived, and increased in short-lived, mutants. These findings reveal that regulators of both PCD and the aging program are required for germline mtDNA quality control and its intergenerational transmission

Regulation of defective mitochondrial DNA accumulation and transmission in C. elegans by the programmed cell death and aging pathways

The heteroplasmic state of eukaryotic cells allows for cryptic accumulation of defective mitochondrial genomes (mtDNA). ‘Purifying selection’ mechanisms operate to remove such dysfunctional mtDNAs. We found that activators of programmed cell death (PCD), including the CED-3 and CSP-1 caspases, the BH3-only protein CED-13, and PCD corpse engulfment factors, are required in C. elegans to attenuate germline abundance of a 3.1-kb mtDNA deletion mutation, uaDf5, which is normally stably maintained in heteroplasmy with wildtype mtDNA. In contrast, removal of CED-4/Apaf1 or a mutation in the CED-4-interacting prodomain of CED-3, do not increase accumulation of the defective mtDNA, suggesting induction of a non-canonical germline PCD mechanism or non-apoptotic action of the CED-13/caspase axis. We also found that the abundance of germline mtDNAuaDf5 reproducibly increases with age of the mothers. This effect is transmitted to the offspring of mothers, with only partial intergenerational removal of the defective mtDNA. In mutants with elevated mtDNAuaDf5 levels, this removal is enhanced in older mothers, suggesting an age-dependent mechanism of mtDNA quality control. Indeed, we found that both steady-state and age-dependent accumulation rates of uaDf5 are markedly decreased in long-lived, and increased in short-lived, mutants. These findings reveal that regulators of both PCD and the aging program are required for germline mtDNA quality control and its intergenerational transmission

Bedtime Salivary Cortisol as a Screening Test for Cushing Syndrome in Children

Background Diagnosing Cushing syndrome (CS) can be challenging. The 24-hour urine free cortisol (UFC) measurement is considered gold standard. This is a laborious test, dependent on correct urine collection. Late-night salivary cortisol is easier and is used as a screening test for CS in adults, but has not been validated for use in children. Objective To define liquid chromatography tandem mass spectrometry (LC-MS/MS)-based cutoff values for bedtime and morning salivary cortisol and cortisone in children, and validate the results in children with and without CS. Methods Bedtime and morning salivary samples were collected from 320 healthy children aged 4 to 16 years. Fifty-four patients from the children’s outpatient obesity clinic and 3 children with pituitary CS were used for validation. Steroid hormones were assayed by LC-MS/MS. Cutoff levels for bedtime salivary cortisol and cortisone were defined by the 97.5% percentile in healthy subjects. Results Bedtime cutoff levels for cortisol and cortisone were 2.4 and 12.0 nmol/L, respectively. Applying these cutoff levels on the verification cohort, 1 child from the obesity clinic had bedtime salivary cortisol exceeding the defined cutoff level, but normal salivary cortisone. All 3 children with pituitary CS had salivary cortisol and cortisone far above the defined bedtime cutoff levels. Healthy subjects showed a significant decrease in salivary cortisol from early morning to bedtime. Conclusions We propose that bedtime salivary cortisol measured by LC-MS/MS with a diagnostic threshold above 2.4 nmol/L can be applied as a screening test for CS in children. Age- and gender-specific cutoff levels are not needed

Bedtime Salivary Cortisol as a Screening Test for Cushing Syndrome in Children

Abstract Background:Diagnosing Cushing syndrome (CS) can be challenging. The 24-hour urine free cortisol (UFC) measurement is considered gold standard. This is a laborious test, de-pendent on correct urine collection. Late-night salivary cortisol is easier and is used as a screening test for CS in adults, but has not been validated for use in children. Objective:To define liquid chromatography tandem mass spectrometry (LC-MS/MS)-based cutoff values for bedtime and morning salivary cortisol and cortisone in children, and validate the results in children with and without CS. Methods:Bedtime and morning salivary samples were collected from 320 healthy chil-dren aged 4 to 16 years. Fifty-four patients from the children’s outpatient obesity clinic and 3 children with pituitary CS were used for validation. Steroid hormones were as-sayed by LC-MS/MS. Cutoff levels for bedtime salivary cortisol and cortisone were de-fined by the 97.5% percentile in healthy subjects. Results:Bedtime cutoff levels for cortisol and cortisone were 2.4 and 12.0 nmol/L, re-spectively. Applying these cutoff levels on the verification cohort, 1 child from the obesity clinic had bedtime salivary cortisol exceeding the defined cutoff level, but normal sal-ivary cortisone. All 3 children with pituitary CS had salivary cortisol and cortisone far above the defined bedtime cutoff levels. Healthy subjects showed a significant decrease in salivary cortisol from early morning to bedtime