Evaluation of the safety of hair essence containing 0.05% purified bee venom on the skin and eyes of rabbits.

OBJECTIVE: To investigate the safety of hair essence containing 0.05% purified bee venom (HE-PBV) on the skin and eyes of New Zealand White rabbits. METHODS: HE-PBV which contained 0.05% PBV, purified water, and glycerin, was used as the test substance. The skin-irritation test (SIT) and eye-irritation test (EIT) were conducted according to the Draize method. On the SIT, HE-PBV (0.5 mL) dropped gauze was attached both intact and abraded skin for 24 h. The other side of the skin was used as control. After 24 and 72 h, the treatment site was observed and scored according to evaluation criteria for skin reactions. On the EIT, the rabbits were divided into two groups: eye-washed (three rabbits) and non-eye-washed (six rabbits). HE-PBV (0.1 mL) was squirted into the right eye of rabbits. The left eye was untreated and used as a control. Then, 20-30-s later, the eyes of rabbits in the eye-washed group were washed with ~50 mL of physiologic (0.9%) salt solution. Then, 1, 2, 3, 4 and 7 d after the start of the EIT, the eyes and behavior of the rabbits were observed. The degree of eye irritation elicited by HE-PBV was determined in three steps and then the criteria of the classification of eye-irritation scores. RESULTS: The SIT revealed erythema and edema at the site of HE-PBV application. At 72 h, the body weight of rabbits was reduced slightly, but other symptoms (except erythema and edema) were not observed. The Primary Irritation Index score was 0.6, and HE-PBV was deemed to be a slight irritant. The EIT did not show mortality or body-weight fluctuation, but hyperemic conjunctiva and eyelid closure were noted after HE-PBV administration. Except for these results, the score for the ophthalmic response on days 1, 2, 3, 4 and 7 was 0, and HE-PBV was deemed to be a non-irritant. CONCLUSION: These data suggest that HE-PBV did not elicit eye irritation, but was a slight irritant to the skin of rabbits; the latter slight would have been due to the excipients used in manufacture of the hair essence because PBV has been shown to be safe.

Translational outcomes in a full gene deletion of ubiquitin protein ligase E3A rat model of Angelman syndrome.

Angelman syndrome (AS) is a rare neurodevelopmental disorder characterized by developmental delay, impaired communication, motor deficits and ataxia, intellectual disabilities, microcephaly, and seizures. The genetic cause of AS is the loss of expression of UBE3A (ubiquitin protein ligase E6-AP) in the brain, typically due to a deletion of the maternal 15q11-q13 region. Previous studies have been performed using a mouse model with a deletion of a single exon of Ube3a. Since three splice variants of Ube3a exist, this has led to a lack of consistent reports and the theory that perhaps not all mouse studies were assessing the effects of an absence of all functional UBE3A. Herein, we report the generation and functional characterization of a novel model of Angelman syndrome by deleting the entire Ube3a gene in the rat. We validated that this resulted in the first comprehensive gene deletion rodent model. Ultrasonic vocalizations from newborn Ube3am-/p+ were reduced in the maternal inherited deletion group with no observable change in the Ube3am+/p- paternal transmission cohort. We also discovered Ube3am-/p+ exhibited delayed reflex development, motor deficits in rearing and fine motor skills, aberrant social communication, and impaired touchscreen learning and memory in young adults. These behavioral deficits were large in effect size and easily apparent in the larger rodent species. Low social communication was detected using a playback task that is unique to rats. Structural imaging illustrated decreased brain volume in Ube3am-/p+ and a variety of intriguing neuroanatomical phenotypes while Ube3am+/p- did not exhibit altered neuroanatomy. Our report identifies, for the first time, unique AS relevant functional phenotypes and anatomical markers as preclinical outcomes to test various strategies for gene and molecular therapies in AS.

Adverse Childhood Experiences and Health and Wellness Outcomes among Black Men Who Have Sex with Men.

Black men who have sex with men (BMSM) are a population at the intersection of two minority statuses-racial minority and sexual minority. Membership in either group, compared to white or heterosexual group membership, may increase one's risk of negative childhood and adult experiences. Baseline data from an HIV intervention efficacy trial (the Black Men Evolving Study) were used to explore the prevalence of adverse childhood experiences (ACEs) among 536 BMSM and associations between ACEs and adult mental and physical health outcomes. Overall, the prevalence of ACEs was high among this sample of BMSM with almost 90% experiencing at least one ACE. Findings revealed that ACE score was significantly associated with adult mental health (AOR = 1.21, 95% CI [1.12, 1.30]), but not with adult physical health. All ACEs were significantly associated with mental health, but only physical neglect and household substance abuse were significantly associated with physical health (AOR = 1.69, 95% CI [1.02, 2.74] and AOR = 1.57, 95% CI [1.03, 2.40], respectively). The findings support the need for interventions targeting improved adult health outcomes, particularly for minority groups, to consider the impact of early adversity on health and wellness.

Ras oncogene-independent activation of RALB signaling is a targetable mechanism of escape from NRAS(V12) oncogene addiction in acute myeloid leukemia.

Somatic mutations that lead to constitutive activation of NRAS and KRAS proto-oncogenes are among the most common in human cancer and frequently occur in acute myeloid leukemia (AML). An inducible NRAS(V12)-driven AML mouse model has established a critical role for continued NRAS(V12) expression in leukemia maintenance. In this model genetic suppression of NRAS(V12) expression results in rapid leukemia remission, but some mice undergo spontaneous relapse with NRAS(V12)-independent (NRI) AMLs providing an opportunity to identify mechanisms that bypass the requirement for Ras oncogene activity and drive leukemia relapse. We found that relapsed NRI AMLs are devoid of NRAS(V12) expression and signaling through the major oncogenic Ras effector pathways, phosphatidylinositol-3-kinase and mitogen-activated protein kinase, but express higher levels of an alternate Ras effector, Ralb, and exhibit NRI phosphorylation of the RALB effector TBK1, implicating RALB signaling in AML relapse. Functional studies confirmed that inhibiting CDK5-mediated RALB activation with a clinically relevant experimental drug, dinaciclib, led to potent RALB-dependent antileukemic effects in human AML cell lines, induced apoptosis in patient-derived AML samples in vitro and led to a 2-log reduction in the leukemic burden in patient-derived xenograft mice. Furthermore, dinaciclib potently suppressed the clonogenic potential of relapsed NRI AMLs in vitro and prevented the development of relapsed AML in vivo. Our findings demonstrate that Ras oncogene-independent activation of RALB signaling is a therapeutically targetable mechanism of escape from NRAS oncogene addiction in AML.

Pre-analytical and analytical variables affecting the measurement of plasma-derived microparticle tissue factor activity.

INTRODUCTION: Elevated levels of tissue factor positive (TF(+)) microparticles (MPs) are observed in plasma from a variety of patients with an increased risk of thrombosis. We and others have described the measurement of TF activity in MPs isolated from plasma. The aim of this study was to investigate the effects of pre-analytical and analytical variables on TF activity of MPs isolated from blood of healthy volunteers either untreated or treated ex vivo with bacterial lipopolysaccharide. MATERIALS AND METHODS: We evaluated the following parameters: use of different centrifugation speeds to isolate the MPs; comparison of TF activity of MPs isolated from platelet poor plasma versus platelet free plasma; effect of freeze/thaw on MP TF activity; and comparison of the MP TF activity assay with the measurement of TF protein by ELISA or flow cytometry. RESULTS: MPs prepared from platelet poor plasma by centrifugation at 20,000×g or 100,000×g for 15 minutes had similar levels of TF activity. However, significantly less TF activity was found in MPs isolated from platelet free plasma compared with platelet poor plasma. Interestingly, freeze/thawing of the plasma showed donor to donor variation in MP TF activity, with a moderate increase in some individuals. CONCLUSION: TF(+) MPs can be quantitatively isolated from platelet poor or platelet free plasma by centrifugation at 20,000×g for 15 minutes. Measurement of MP TF activity in plasma may be used to detect a prothrombotic state in patients with various diseases.

The effect of time-of-day dosing on the pharmacokinetics and pharmacodynamics of dexlansoprazole MR: evidence for dosing flexibility with a Dual Delayed Release proton pump inhibitor.

BACKGROUND: Dexlansoprazole MR is a Dual Delayed Release proton pump inhibitor formulated to extend the duration of acid suppression. AIM: To evaluate the pharmacokinetics and pharmacodynamics of dexlansoprazole MR dosed before 4 different meal times. METHODS: In this randomized, open-label, four-way crossover study, 48 healthy subjects received dexlansoprazole MR 60 mg once daily 30 min before breakfast, lunch, dinner or an evening snack. Pharmacokinetics of dexlansoprazole MR and intragastric pH were assessed over a 24-h postdose interval on day 5 for each regimen. RESULTS: Absorption was delayed when dexlansoprazole MR was administered before each regimen relative to breakfast; however, systemic exposures of dexlansoprazole at all regimens were bioequivalent. There were no statistically significant differences in mean 24-h intragastric pH between dosing before dinner or an evening snack vs. breakfast; however, there was a small (0.2), but statistically significant difference between lunch and breakfast. There was a statistically significant difference of 7 percentage points in the percentage of time intragastric pH was >4 for the snack regimen relative to the breakfast regimen, but there were no statistically significant differences between lunch or dinner compared with breakfast. CONCLUSION: Dexlansoprazole MR provides comparable acid control when administered at different times of the day.

Clinical trial: the effect and timing of food on the pharmacokinetics and pharmacodynamics of dexlansoprazole MR, a novel Dual Delayed Release formulation of a proton pump inhibitor--evidence for dosing flexibility.

BACKGROUND: Dexlansoprazole MR is a proton pump inhibitor with a Dual Delayed Release (DDR) formulation designed to prolong the dexlansoprazole plasma concentration-time profile. The presence of food or time of dosing relative to food may affect dexlansoprazole absorption. AIMS: To evaluate the effect of food on the pharmacokinetics (PK) and pharmacodynamics (PD) of dexlansoprazole following oral administration of dexlansoprazole MR. METHODS: In this open-label, single-dose, randomized, 4-way crossover study, 48 healthy subjects received placebo (day 1) and dexlansoprazole MR 90 mg (day 3) after fasting, 5 or 30 min before a high-fat breakfast, or 30 min after a high-fat breakfast. Intragastric pH (days 1 and 3) and PK (day 3) of dexlansoprazole were assessed over a 24-h interval after each dose. RESULTS: Following administration of dexlansoprazole MR under fasted/fed conditions, mean dexlansoprazole plasma concentration-time profiles generally exhibited two distinct peaks, resulting from the DDR formulation. Increases in dexlansoprazole maximum plasma concentration (12-31%) and area under the plasma concentration-time curve (9-21%) were observed with the fed regimens; however, differences in intragastric pH were not considered clinically relevant. CONCLUSION: Dexlansoprazole MR can be administered without regard to food or the timing of food in most patients.

Identification of Transcripts Involved in Resistance Responses to Leaf Spot Disease Caused by Cercosporidium personatum in Peanut (Arachis hypogaea).

ABSTRACT Late leaf spot disease caused by Cercosporidium personatum is one of the most destructive foliar diseases of peanut (Arachis hypogaea) worldwide. The objective of this research was to identify resistance genes in response to leaf spot disease using microarray and real-time polymerase chain reaction (PCR). To identify transcripts involved in disease resistance, we studied the gene expression profiles in two peanut genotypes, resistant or susceptible to leaf spot disease, using cDNA microarray containing 384 unigenes selected from two expressed sequenced tag (EST) cDNA libraries challenged by abiotic and biotic stresses. A total of 112 spots representing 56 genes in several functional categories were detected as up-regulated genes (log(2) ratio > 1). Seventeen of the top 20 genes, each matching gene with known function in GenBank, were selected for validation of their expression levels using real-time PCR. The two peanut genotypes were also used to study the functional analysis of these genes and the possible link of these genes to the disease resistance trait. Microarray technology and real-time PCR were used for comparison of gene expression. The selected genes identified by microarray analysis were validated by real-time PCR. These genes were more greatly expressed in the resistant genotype as a result of response to the challenge of C. personatum than in the susceptible genotype. Further investigations are needed to characterize each of these genes in disease resistance. Gene probes could then be developed for application in breeding programs for marker-assisted selection.

Mapping microbial biodiversity.

We report the development of a prototype database that "maps" microbial diversity in the context of the geochemical and geological environment and geographic location. When it is fully implemented, scientists will be able to conduct database searches, construct maps containing the information of interest, download files, and enter data over the Internet.

Should all infertile males undergo urologic evaluation before assisted reproductive technologies? Two cases of testicular cancer presenting with infertility.

OBJECTIVE: To report two cases of testicular cancer in patients presenting with infertility. DESIGN: Case reports. SETTING: University-affiliated urology practice. PATIENT(S): Two men presenting with infertility. INTERVENTION(S): Complete history and physical, hormonal assays, semen analysis, scrotal ultrasound, radical orchiectomy. MAIN OUTCOME MEASURE(S): Testicular pathology specimens. RESULT(S): Testicular cancer was diagnosed in two men sent to a urology clinic for infertility treatment. CONCLUSION(S): A thorough evaluation should be completed in all males in couples presenting with infertility.

Effects of ochratoxin A on cytotoxicity and cell differentiation in cultured rat embryonic cells.

In the present study, the effects of ochratoxin A (OTA) on cytotoxicity, cell differentiation, and other cell functions in the embryonic midbrain cells, which are dopaminergic, were compared to those in the limb bud cells, which are nondopaminergic, to assess the selectivity of OTA central action. Twelve-day rat embryo midbrain and limb bud cells were cultured in Dulbecco's modified Eagle's medium nutrient and Ham's F12 (1:1) mix ture containing 10% Nuserum for 96 h in the presence of various concentrations of OTA. OTA signicfiantly reduced the levels of protein, DNA and glutathione, and [H]thymidine incorporation into DNA in both embryonic midbrain and limb bud cells in a similar concentration-dependent manner. The IC50 values for cytotoxicity measured by neutral red uptake were 1.10 microM in the midbrain cells and 1.05 microM in the limb bud cells. The IC50 values of cell differentiation were 1.10 microM in the midbrain cells and 1.0 microM in the limb bud cells. The addition of exogenous glutathione (32.5 microM) did not change the OTA-induced fall in protein and DNA levels, or the IC50 values of cytotoxicity and differentiation in the midbrain and limb bud cells. Data show that OTA does not appear to exert a selective toxic dopaminergic cell action and that OTA-induced cytotoxicity and inhibition of cell differentiation were not prevented by exogenous glutathione.

In vitro metabolism of the HIV-1 protease inhibitor ABT-378: species comparison and metabolite identification.

HIV protease inhibitor ABT-378 (ABT-378) was metabolized very extensively and rapidly by liver microsomes from mouse, rat, dog, monkey, and humans. The rates of NADPH-dependent metabolism of ABT-378 ranged from 2.39 to 9.80 nmol.mg microsomal protein-1.min-1, with monkey liver microsomes exhibiting the highest rates of metabolism. ABT-378 was metabolized to 12 metabolites (M-1 to M-12), which were characterized by mass and NMR spectroscopy. The metabolite profile of ABT-378 in liver microsomes from all five species was similar, except that the mouse liver microsomes did not form M-9, a minor secondary metabolite. The predominant site of metabolism was the cyclic urea moiety of ABT-378. In all five species, the major metabolites were M-1 (4-oxo-ABT-378) and M-3 and M-4 (4-hydroxy-ABT-378). Metabolite M-2 (6-hydroxy-ABT-378) was formed by rodents at a faster rate than by dog, monkey, and human liver microsomes. Metabolites M-5 to M-8 were identified as monohydroxylated derivatives of ABT-378. Metabolites M-9 and M-10 were identified as hydroxylated products of M-1. Metabolites M-11 and M-12 were identified as dihydroxylated derivatives of ABT-378. The metabolite profile in human hepatocytes and liver slices was similar to that of human liver microsomes. The results of the current study indicate that ABT-378 is highly susceptible to oxidative metabolism in vitro, and possibly in vivo, in humans.

Neither dopamine nor dobutamine corrects mesenteric blood flow depression caused by positive end-expiratory pressure in a rat model of acute lung injury.

OBJECTIVE: To determine if either dopamine or dobutamine would counteract the deleterious effect that positive end-expiratory pressure (PEEP) has on cardiac output and mesenteric blood flow in a rat model of acute lung injury. DESIGN: Prospective, randomized, controlled trial in a clinically relevant model of acute lung injury. SETTING: Microcirculation research laboratory. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: The animals were anesthetized with pentobarbital (30 mg/kg) by intraperitoneal injection. They underwent tracheostomy, jugular and femoral vein cannulation, femoral artery cannulation, carotid artery thermistor placement, and bowel preparation for in vivo video microscopy. Acute lung injury was created by administering 0.1 N hydrochloric acid (1 mL/kg) via the tracheostomy. Dopamine or dobutamine (2.5 or 12.5 microg/kg/min), followed by two intravenous fluid boluses, was administered to rats ventilated with 5, 10, 15, and 20 cm H2O of PEEP. MEASUREMENTS AND MAIN RESULTS: Mean arterial pressure, thermodilution cardiac output, mesenteric arteriolar diameter, and red blood cell velocity were measured and mesenteric blood flow was calculated. Cardiac output was depressed in rats exposed to 20 cm H2O of PEEP by 32+/-2%. The corresponding values for cardiac output depression at 20 cm H2O of PEEP in rats receiving 2.5 and 12.5 microg/kg/min of dopamine and 2.5 and 12.5 microg/kg/min of dobutamine were 31+/-1%, 21+/-1%, 29+/-0%, and 24+/-2%, respectively. Mesenteric blood flow was depressed in rats ventilated with 20 cm H2O of PEEP by 74+/-3%, while the corresponding values in rats exposed to 20 cm H2O of PEEP and receiving 2.5 or 12.5 microg/kg/min of dopamine or 2.5 or 12.5 microg/kg/min of dobutamine were 86+/-3%, 77+/-3%, 73+/-3%, and 66+/-3%, respectively. Fluid boluses did not correct the deficits in cardiac output or mesenteric blood flow caused by the combination of acute lung injury and PEEP. CONCLUSIONS: The higher doses of dopamine and dobutamine partially, but insignificantly, corrected the cardiac output depression caused by PEEP in a model of acute lung injury. Neither dose of dopamine nor dobutamine was able to improve PEEP-induced mesenteric blood flow depression.

Polypharmacy: a case report and new protocol for management.

BACKGROUND: Polypharmacy is an important issue in primary care, yet few data are available concerning its prevalence, complications, and management in clinical medicine. The following case illustrates the clinical perils of polypharmacy and serves as a point for critical discussion. METHODS: MEDLINE was searched, using the key word "polypharmacy," from 1994 to the present. A case report of polypharmacy is described, and a novel protocol for the management of polypharmacy is proposed. RESULTS: Polypharmacy can lead to unnecessary expense, wasted time, and embarrassment on the part of the patient and confusion and mismanagement on the part of the physician. The literature reveals controversy surrounding the definition of polypharmacy and reflects the considerable morbidity and expense associated with polypharmacy. Finally, the SAIL protocol shows that physicians need to keep in mind simplicity, adverse effects, indications, and a precise list of all medications to manage appropriately a patient's drug regimen. CONCLUSIONS: Polypharmacy is associated with morbidity and iatrogenic complications. The SAIL protocol can be a useful tool in the management of this entity. More research needs to be done on the prevalence, complications, and management of polypharmacy.

Effects of severe hypoxemia on mesenteric blood flow in neonatal piglets.

BACKGROUND: The purpose of this work was to determine the effects of hypoxemia on systemic hemodynamic variables and regional conduit arterial blood flows in neonatal piglets. MATERIALS AND METHODS: Using transit time blood flow probes, cardiac output and cranial mesenteric artery blood flow were monitored in groups of prematurely delivered (90% of term gestational age) and 2-week-old piglets prior to, during, and after exposure to a 30-min hypoxic (FIO2 = 0.12) challenge. RESULTS: The documented alterations in systemic mean arterial pressure and cardiac output pressure during hypoxia and reoxygenation were consistent with the maturational age of the animals. In response to hypoxia, all animals demonstrated significant reductions in mesenteric blood flow, with returns to baseline levels during the 30-min reoxygenation phase. In still other prematurely delivered piglets, nutrient mesenteric arterial blood flows were measured using in vivo videomicroscopy. The marked hypoxemia-induced decline in mucosal blood flow was reversed by reoxygenation. CONCLUSIONS: The physiologic mechanisms responsible for neonatal mesenteric vasoactive responsiveness are present in conduit and in nutrient vessels well prior to birth and can be activated by a significant perturbation. These observations are germane insofar as they provide a stable, age-matched acute animal model to study neonatal intestinal ischemic diseases, including necrotizing enterocolitis.

Urban versus rural: fertility decline in the cities and rural districts of Prussia, 1875 to 1910.

PIP: This study examined the level and pace of fertility decline in 54 large cities in Prussia during 1875-1910. Data were obtained from census records for 1875, 1880, 1885, 1890, 1895, 1900, 1905, and 1910. Analysis was based on pooled cross sectional time series methods and theories that structural socioeconomic change was a key factor in the decline of fertility in Prussian cities. During 1875-1910, city population grew from 3.8 to 9.6 million. The general marital fertility rate (GMFR) in the 54 cities declined from 281 to 164 during 1875-1910. Catholicism, female labor force participation (FLFP), manufacturing, mining, banking, and communication were statistically positively related to urban fertility level. Pace of decline was related significantly to language, education, FLFP, income, communications, insurance, population size, infant mortality, and married sex ratio. Population size was related to pace but not level. Education and banking had a stronger impact in rural areas. In the city equation, the variables plus dummies accounted for 90% of the variance. A very important variable explaining change in fertility level in Prussia was Catholicism. The most important variables for explaining fertility change in Prussia were infant mortality rate and insurance. Infant mortality, communications, insurance, and income increased in importance over time. FLFP declined over time, but contributed the most to predicted change in GMFR throughout the period, especially in nontraditional occupations. The analysis explained both rapid and slow urban fertility declines and closely approximated predicted fertility. Prussia differs from present developing country contexts in that the population was largely agrarian but literate.^ieng

Vitamin A, visual pigments, and visual receptors in Drosophila.

The fly visual system has served for decades as a model for receptor spectral multiplicity and vitamin A utilization. A diverse armamentarium of structural techniques has dovetailed with convenient electrophysiology, photochemistry, genetics, and molecular biology in Drosophila to facilitate recent progress, which is reviewed here. New data are also presented. Ultrastructure of retinula cells of carotenoid-deprived flies shows that organelles associated with protein biosynthesis, i.e., rough endoplasmic reticulum and Golgi apparatus, are present, while organelles associated with rhabdomere turnover, i.e., multivesicular bodies (MVBs), are rare. Ultrastructure and morphometry suggest that retinoic acid-rearing stimulates membrane export and rhabdomere buildup, even though functional rhodopsin is missing. Confocal microscopy suggests that RH4, one of the ultraviolet rhodopsins, may reside in the previously-described pale fluorescent R7 cells with RH3 in the yellow fluorescent R7 cells.

Visualization of iontophoretic transport paths in cultured and animal skin models.

Physiological structures associated with iontophoretic paths in hairless mouse skin and two cultured skin models ("EpiDerm" by Mattek, Corp., and "SKIN2" by Advanced Tissue Sciences, Inc.) are reported. Visualization of ionic paths at current densities between 20 and 100 microA/ cm2 is accomplished by the counterdirectional transport of Fe(CN)6(4-) and Fe3+, resulting in the controlled precipitation of colloidal Prussian blue, Fe4[Fe(CN)6]3, at sites of high ionic conductivity. Examination of the Fe4[Fe(CN)6]3-stained tissues using optical microscopy allows unequivocal assignment of iontophoretic paths to physiological structures in the stratum corneum. Deposition of Fe4[Fe(CN)6]3 occurs exclusively at hair follicles in hairless mouse skin, indicating that these appendages provide highly conductive porous paths during iontophoresis. In contrast, the counterdirectional transport of Fe(CN)6(4-) and Fe3+ across cultured skin models, which lack appendages, results in the deposition of Fe4-[Fe(CN)6]3 along the boundaries of corneocytes. This observation suggests that paracellular iontophoretic transport through lipid bilayer regions is the predominant transport path in the absence of low-resistance pores.

Control of Drosophila opsin gene expression by carotenoids and retinoic acid: northern and western analyses.

In the fly, thorough retinoid deprivation is possible, to optimize investigation of the effects of vitamin A metabolites and retinoic acid (RA) on visual development. Retinoids had been found to control fly opsin gene transcription, though this finding was contested. Northern blots on Drosophila heads showed that mRNA of Rh1 (the predominant rhodopsin) was high in vitamin A replete controls, very low in deprived flies, and increased upon feeding carrot juice to deprived flies as early as 1 hr. Expression of the ribosomal protein 49 [rp49] gene (the control) was equal both in deprivation and in replacement. Recovery of Rh1 protein upon such carotenoid replacement followed, barely detectable on Western blots at 4 hr but conspicuous by 8 hr. Alternative chromophore deprivation with yeast-glucose food yielded flies with opsin mRNA on Northerns but not rhodopsin, as demonstrated by Western blots, spectrophotometry and the electroretinogram (ERG). Rh1's mRNA but not Rh1 protein resulted from rearing flies from egg to adult on the otherwise deprivational medium supplemented with RA or beef brain-heart infusion. By comparing results from these different media it was concluded that: [1] deprivation and replacement affect opsin gene transcription; and [2] contradictory conclusions were from chromophore deprivation which does not eliminate all retinoid dependent factors which could affect the opsin promoter. Preliminary evidence shows that carotenoid deprivation decreases two proteins relevant to visual function: [1] phospholipase C (PLC); and [2] Drosophila retinoid binding protein (DRBP).