A follow-up on quantitative and qualitative olfactory dysfunction and other symptoms in patients recovering from COVID-19 smell loss.

BACKGROUND: Sudden smell loss is a specific early symptom of COVID-19, which, prior to the emergence of Omicron, had estimated prevalence of ~40% to 75%. Chemosensory impairments affect physical and mental health, and dietary behavior. Thus, it is critical to understand the rate and time course of smell recovery. The aim of this cohort study was to characterize smell function and recovery up to 11 months post COVID-19 infection. METHODS: This longitudinal survey of individuals suffering COVID-19-related smell loss assessed disease symptoms and gustatory and olfactory function. Participants (n=12,313) who completed an initial survey (S1) about respiratory symptoms, chemosensory function and COVID-19 diagnosis between April and September 2020, were invited to complete a follow-up survey (S2). Between September 2020 and February 2021, 27.5% participants responded (n=3,386), with 1,468 being diagnosed with COVID-19 and suffering co-occurring smell and taste loss at the beginning of their illness. RESULTS: At follow-up (median time since COVID-19 onset ~200 days), ~60% of women and ~48% of men reported less than 80% of their pre-illness smell ability. Taste typically recovered faster than smell, and taste loss rarely persisted if smell recovered. Prevalence of parosmia and phantosmia was ~10% of participants in S1 and increased substantially in S2: ~47% for parosmia and ~25% for phantosmia. Persistent smell impairment was associated with more symptoms overall, suggesting it may be a key marker of long-COVID illness. The ability to smell during COVID-19 was rated slightly lower by those who did not eventually recover their pre-illness ability to smell at S2. CONCLUSIONS: While smell ability improves for many individuals who lost it during acute COVID-19, the prevalence of parosmia and phantosmia increases substantially over time. Olfactory dysfunction is associated with broader persistent symptoms of COVID-19, and may last for many months following acute COVID-19. Taste loss in the absence of smell loss is rare. Persistent qualitative smell symptoms are emerging as common long-term sequelae; more research into treatment options is strongly warranted given that even conservative estimates suggest millions of individuals may experience parosmia following COVID-19. Healthcare providers worldwide need to be prepared to treat post COVID-19 secondary effects on physical and mental health.

Age-related differences in network structure and dynamic synchrony of cognitive control.

Cognitive trajectories vary greatly across older individuals, and the neural mechanisms underlying these differences remain poorly understood. Here, we investigate the cognitive variability in older adults by linking the influence of white matter microstructure on the task-related organization of fast and effective communications between brain regions. Using diffusion tensor imaging and electroencephalography, we show that individual differences in white matter network organization are associated with network clustering and efficiency in the alpha and high-gamma bands, and that functional network dynamics partly explain individual differences in cognitive control performance in older adults. We show that older individuals with high versus low structural network clustering differ in task-related network dynamics and cognitive performance. These findings were corroborated by investigating magnetoencephalography networks in an independent dataset. This multimodal (fMRI and biological markers) brain connectivity framework of individual differences provides a holistic account of how differences in white matter microstructure underlie age-related variability in dynamic network organization and cognitive performance.

Factors associated with glycaemic control in adults with Type 1 diabetes: a registry-based analysis including 7601 individuals from 34 centres in Norway.

AIMS: To use data from the Norwegian Diabetes Registry for Adults and Statistics Norway to assess factors associated with glycaemic control in type 1 diabetes. METHODS: The analyses included all individuals aged ≥18 years who had a type 1 diabetes duration of >2 years and a recorded value in the registry between 2013 and 2015 (n=7601). Predicted mean HbA1c levels for subgroups of participants were assessed using linear regression analysis. RESULTS: Young age (18-25 years), low education levels, smoking, living alone, exercising infrequently, monitoring glucose infrequently, high insulin requirements, low frequency of symptomatic hypoglycaemia, history of ketoacidosis and a BMI <18.5 kg/m2 were associated with a 2-12-mmol/mol (0.2-1.1%) higher HbA1c level. Those with 10-15 years of diabetes duration had 5-mmol/mol (0.5%) higher HbA1c level than those who had a diabetes duration of 2-5 years. Sex, participation (ever) in a diabetes education course, or ever experiencing serious hypoglycaemia were not associated with glycaemic control. CONCLUSIONS: We present representative national data on factors that were associated with glycaemic control. A better understanding and awareness of these factors, together with technological advances in diabetes management, could lead to more personalized management strategies, better glycaemic control and a lower risk of diabetes complications.

Variation in the achievement of HbA1c , blood pressure and LDL cholesterol targets in type 2 diabetes in general practice and characteristics associated with risk factor control.

AIMS: To identify population, general practitioner, and practice characteristics associated with the achievement of HbA1c , blood pressure and LDL cholesterol targets, and to describe variation in the achievement of risk factor control. METHODS: We conducted a cross-sectional survey of 9342 people with type 2 diabetes, 281 general practitioners and 77 general practices in Norway. Missing values (7.4%) were imputed using multiple imputation by chained equations. We used three-level logistic regression with the achievement of HbA1c , blood pressure and LDL cholesterol targets as dependent variables, and factors related to population, general practitioners, and practices as independent variables. RESULTS: Treatment targets were achieved for HbA1c in 64%, blood pressure in 50%, and LDL cholesterol in 52% of people with type 2 diabetes, and 17% met all three targets. There was substantial heterogeneity in target achievement among general practitioners and among practices; the estimated proportion of a GPs diabetes population at target was 55-73% (10-90 percentiles) for HbA1c , 36-63% for blood pressure, and 47-57% for LDL cholesterol targets. The models explained 11%, 5% and 14%, respectively, of the total variation in the achievement of HbA1c , blood pressure and LDL cholesterol targets. Use among general practitioners of a structured diabetes form was associated with 23% higher odds of achieving the HbA1c target (odds ratio 1.23, 95% confidence interval (CI) 1.02-1.47) and 17% higher odds of achieving the LDL cholesterol target (odds ratio 1.17, 95% CI 1.01-1.35). CONCLUSIONS: Clinical diabetes management is difficult, and few people meet all three risk factor control targets. The proportion of people reaching target varied among general practitioners and practices. Several population, general practitioner and practice characteristics only explained a small part of the total variation. The use of a structured diabetes form is recommended.

Population, general practitioner and practice characteristics are associated with screening procedures for microvascular complications in Type 2 diabetes care in Norway.

AIMS: To assess population, general practitioner (GP) and practice characteristics associated with the performance of microvascular screening procedures and to propose strategies to improve Type 2 diabetes care. METHODS: A cross-sectional survey in Norway (281 GPs from 77 practices) identified 8246 people with a Type 2 diabetes duration of 1 year or more. We used multilevel regression models with either the recording of at least two of three recommended screening procedures (albuminuria, monofilament, eye examination) or each procedure separately as dependent variable (yes/no), and characteristics related to the person with diabetes, GP or practice as independent variables. RESULTS: The performance of recommended screening procedures was recorded in the following percentages: albuminuria 31.5%, monofilament 27.5% and eye examination 60.0%. There was substantial heterogeneity between practices, and between GPs within practices for all procedures. Compared with people aged 60-69 years, those aged < 50 years were less likely to have an albuminuria test performed [odds ratio (OR) 0.75, 95% CI 0.61 to 0.93] and eye examination (OR 0.79, 95% CI 0.66 to 0.95). People with macrovascular disease had fewer screening procedures recorded (OR 0.68, 95% CI 0.59 to 0.78). Use of an electronic diabetes form was associated with improved screening  (OR 2.65, 95% CI 1.86 to 3.78). GPs with high workload recorded fewer procedures (OR 0.59, 95% CI 0.39 to 0.90). CONCLUSIONS: Performance of screening procedures was suboptimal overall, and in people who should be prioritized. Performance varied substantially between GPs and practices. The use of a structured diabetes form should be mandatory.

Competition between selenomethionine and methionine absorption in the intestinal tract of green sturgeon (Acipenser medirostris).

l-Selenomethionine (SeMet) is a dominant form of selenium (Se) found in organisms at all levels of aquatic food chains and a key source of Se bioaccumulation and ecotoxicity. In mammals, intestinal absorption of SeMet is at least partly via the Na(+)-dependent neutral amino acid transporter. The mechanism of SeMet absorption and competitive effects of other dietary components on SeMet absorption in fish are unknown. Thus the in vitro uptake rates of l-methionine (Met) and the competitive effect of SeMet on Met absorption, an indicator that SeMet uses the same nutrient transporter(s) for absorption, in the various regions of the green sturgeon (Acipenser medirostris) intestine were investigated using intact tissues (a modified everted sleeve method). Intestinal tissue was incubated in Ringer's solution containing 0-10mmolL(-1) Met or SeMet (n=5 for each substrate's concentration and intestinal region), respectively, as well as constant tracer levels of isotope-labeled Met. The data indicate that SeMet uptake was mediated by the same transporter(s) as Met and that the absorption kinetics were similar for both substrates. When there were differences in absorption they appeared to be mostly due to higher permeability (passive uptake) of the tissue for Met than for SeMet, particularly in the pyloric caeca (PC) and distal intestine (DI). Maximum rates of absorption, on the other hand, tended to be higher for SeMet than Met in the mid intestine and DI, whereas differences in affinity for the transporters varied between these tissues but were very similar in the PC. These differences may be due to differences in regional intestinal characteristics such as amount of mucus secreted and degree of tissue contraction, and/or substrate differences regarding solubility in and movement through the mucus, influence on tissue contraction, permeability through membranes or between cells, intracellular metabolism, as well as basolateral transport. Interestingly, an increasing proximal-to-distal gradient for rates of methionine and putative SeMet absorption was observed in green sturgeon which differs from the declining gradient usually observed for substrate absorption in other fish species and mammals.

Consumer liking of refined and whole wheat breads.

Preference for refined bread is often cited as a reason for the relatively low consumption of whole wheat bread; only a few studies, however, have examined consumer preferences between refined and whole wheat breads, and the results of these studies are inconclusive. Our objective was to determine if refined wheat bread is preferred to whole wheat bread. We hypothesized that people would prefer refined wheat bread. We conducted a taste test with 89 people. They rated their liking of 9 different breads chosen to represent several comparisons between equivalent refined and whole wheat breads. The participants also rated the intensity of 6-n-propylthiouracil (PROP) and completed a questionnaire about their bread preferences and purchasing habits. We classified the participants by their bread preference and their PROP taster status, and then examined the liking patterns of these subgroups. People preferred refined bread to whole wheat bread when both were made using equivalent ingredients and procedures. They liked the commercial samples of refined and whole wheat breads equally well. When people were classified by their bread preference, those who preferred refined bread liked the refined bread better in all comparisons. PROP nontasters liked all refined and whole wheat breads equally. Sensory preferences are a barrier to whole wheat bread consumption, but ingredient or processing modifications can improve liking of whole wheat bread to the level of refined bread.

Prevalence of bladder, bowel and sexual problems among multiple sclerosis patients two to five years after diagnosis.

Most multiple sclerosis (MS) patients experience some sexual, bladder and/or bowel dysfunction during the course of the disease--one of MS most disabling features. This study estimated the frequency of these problems among patients, two to five years after diagnosis, and investigated how these problems are associated with health-related quality of life (using the Multiple Sclerosis Quality of Life-54 questionnaire). The study population comprised a cohort of patients (n = 56), diagnosed in a three-year period, in Hordaland County, Norway. The patients were examined clinically, including scoring of the Expanded Disability Status Scale (EDSS), and completed questionnaires related to bowel and bladder dysfunction, sexual problems and health-related quality of life. More than half the patients had bladder and sexual problems. The frequency of self-reported bladder problems corresponded to the relatively high levels of residual urine found. The presence of these problems was associated with lower scores on the quality of life scales. Further, the bowel problems reported were markedly associated with the quality of life scores. Since treatments and preventive strategies can manage many of these problems, we suggest increasing the focus on these aspects of the disease when consulting patients, including at early stages.

Intestinal stromal tumors in a simian immunodeficiency virus-infected, simian retrovirus-2 negative rhesus macaque (Macaca mulatta).

Multifocal submucosal stromal tumors were diagnosed in a 5.5-year-old rhesus macaque (Macaca mulatta) experimentally infected with simian immunodeficiency virus, strain SIVsmE660, and CD4+ T cell depleted. The animal was negative for simian retroviruses, SRV-1, -2, and -5. Polymerase chain reaction analysis of DNA from tumor and spleen tissue revealed abundant, preferential presence of retroperitoneal fibromatosis herpesvirus, the macaque homologue of the Kaposi sarcoma-associated herpesvirus (human herpesvirus-8), in the tumors. This was corroborated by demonstration of viral latent nuclear antigen-1 in the nuclei of a majority of the spindeloid tumor cells. Low levels of an additional macaque herpesvirus, rhesus rhadinovirus, were also detected in the spleen and tumor tissues. The spindeloid cells labeled positively for vimentin and CD117 but were negative for CD31, CD68, desmin, and smooth muscle cell actin. Collectively, these findings suggest a relation to but not absolute identity with simian mesenchymoproliferative disorders (MPD) or typical gastrointestinal stromal tumors (GISTs).

Prospective immunological profiling in a case of immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX).

IPEX syndrome is a genetic autoimmune disease characterized by immune-mediated polyendocrinopathy, enteropathy, and X-linked inheritance. We describe a case of IPEX in which lymphocyte phenotypes were assessed at birth, before initiation of Cyclosporin A therapy, and at frequent intervals to 18 months of age. We performed flow cytometry for lymphocyte subtypes and for activation markers (HLA-DR, CD25, and CD69 or CD71). The ratios of both T to B cells and CD4+ to CD8+ cells were elevated at birth, but CD4+ cells were not activated. HLA-DR+ and CD25+ activated T-cells increased in association with two episodes of clinical deterioration: colitis and the onset of type I diabetes mellitus. These results indicate that measures of activation, particularly HLA-DR+ and CD25+ frequency, correlate well with the development of early active disease and may presage clinical episodes. Continuous maintenance of immunosuppression, once started, appears critical for prevention of permanent tissue damage.

Rescue of the autoimmune scurfy mouse by partial bone marrow transplantation or by injection with T-enriched splenocytes.

The scurfy mutant mouse is the genetic and phenotypic equivalent of the single-gene human autoimmune disease immunodysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX). The scurfy mutation disrupts the Foxp3 gene, a putative master switch for T regulatory cell development. Bone marrow transplant without conditioning was previously reported to be ineffective in scurfy mice, yet clinical remission occurs in transplanted human IPEX patients despite limited donor engraftment. In view of this contradiction, we sought to validate scurfy as a model for studying the pathogenesis and treatment of human IPEX, in particular the phenomenon of dominant immune regulation. One half of scurfy mice given bone marrow transplants after sublethal irradiation recovered and survived long-term with donor chimerism ranging from 1.7% to 50%. Early transfer of 2 x 107 normal T cell-enriched splenocytes also prevented or limited disease and permitted long-term survival. Donor T cells in rescued mice made up 3-5% of lymphocytes and became highly enriched for CD25+ T cells over time. Transfer of 106 CD4+ CD25+ sorted T cells showed some beneficial effect, while CD4+ CD25- cells did not. Thus, both partial bone marrow transplant and T-enriched splenocyte transfer are effective treatments for scurfy. These results indicate that scurfy results from a lack of cells with dominant immune regulatory capacity, possibly T regulatory cells. The potency of small numbers of normal cells indicates that IPEX may be a feasible target for gene therapy.

Neutrophil CD64 expression: distinguishing acute inflammatory autoimmune disease from systemic infections.

BACKGROUND: Common bacterial and opportunistic infections are a major cause of mortality in patients who are immunosuppressed owing to treatment with corticosteroids or cytotoxic drugs. Common laboratory tests for infection lack sensitivity and specificity. One of the new generation of tests to detect early systemic infections measures the up regulation of an Fc receptor (Fcgamma R1, or CD64) on neutrophils. The Fc receptors on white blood cells are very important for effective phagocytosis of bacteria and are up regulated during an infection. OBJECTIVE: To measure the clinical usefulness of quantitative CD64 measurements to differentiate between systemic infection and active autoimmune inflammation in an ongoing study. METHODS: Patients with systemic infection (n=27), active autoimmune inflammatory disease (n=44), vasculitis (n=5), and controls (n=20) were studied for neutrophil CD64 expression using monoclonal antibodies and flow cytometry. RESULTS: The median (interquartile range (IQR)) CD64 expression in patients with active inflammatory disease and systemic infection was 907.5 (586-1550) and 3647 (2380-6642), respectively (p<0.0001). The median (IQR) CD64 expression in control patients (osteoarthritis and fibromyalgia) was 505 (359-599). The sensitivity and specificity of CD64 expression on neutrophils to diagnose systemic infection (using a cut off value of 2000) was 85% and 91%, respectively. CONCLUSION: These results indicate that quantitative measurement of CD64 can distinguish between systemic infection and the flare of autoimmune diseases.

Diminished frequency of interleukin-10-secreting, T-cell receptor peptide-reactive T cells in multiple sclerosis patients might allow expansion of activated memory T cells bearing the cognate BV gene.

T cells responsive to T-cell receptor (TCR) determinants may regulate pathogenic Th1 responses in patients with multiple sclerosis (MS) through interleukin (IL)-10-dependent bystander suppression. In this study, innate IL-10- and interferon (IFN)-gamma-secreting T cells responsive to TCR peptides were quantified in peripheral blood mononuclear cells of MS patients and healthy controls (HC) using the ELISPOT assay. Most HC had vigorous IL-10 but low IFN-gamma frequencies to BV5S2 and BV6S1 peptides. In contrast, MS patients had significantly lower IL-10 frequency responses to the TCR peptides but normal responses to concanavalin A. Patients undergoing TCR-peptide vaccination had moderate responses that fluctuated in concert with vaccination. In an MS patient and HC, expression of BV6S1 by activated memory T cells was inversely associated with the presence of IL-10-secreting BV6S1-reactive T cells. These results suggest that MS patients have diminished frequencies of innate TCR-reactive T cells that may allow oligoclonal expansion of activated autoreactive Th1 effector cells expressing cognate V genes.

Reduced quality of life among multiple sclerosis patients with sexual disturbance and bladder dysfunction.

OBJECTIVE: Physical disability explains only part of the reduced quality of life found among multiple sclerosis (MS) patients. Bladder dysfunction and sexual disturbance are frequent and distressing problems for MS patients. We therefore estimated the relationship between the presence and degree of sexual disturbance/bladder dysfunction and the patients' quality of life as measured by the SF-36 Health Survey. METHODS: We performed a cross-sectional study of all individuals with the onset of MS between 1976 and 1986 in Hordaland County, Norway. The disease duration at examination was 9-19 years; 194 patients (94%) participated. RESULTS: Fifty-three per cent of the patients with low physical disability (Expanded Disability Status Scale (EDSS) < or = 4.0) reported disease-related sexual disturbance and 44% had bladder dysfunction according to the Incapacity Status Scale. The corresponding figures for the patients with a high physical disability (EDSS > 4.0) were 86 and 81% respectively. The patients with sexual disturbance had markedly and significantly reduced scores on all eight SF-36 scales, this was after adjustment for disease development measured by the EDSS. The patients without sexual disturbance scored 0.5 s.d. lower than a normal population on the social functioning scale, whereas those with marked sexual disturbance scored 1.5 s.d. lower. Similar results were found for the patients with bladder dysfunction. CONCLUSION: Bladder and sexual problems are associated with a marked reduction in the quality of life, also among patients with otherwise low disability. This underlines the need for identifying and treating these problems.

Reconstructive urology in the nordic countries--a hospital questionnaire survey.

A hospital survey of adult reconstructive urologic surgery in the Nordic countries is presented. The response rate was 80% and included most general hospitals and university clinics. Despite similarities between the healthcare systems of the various countries several differences were found. Cystectomy was performed in a large number of institutions in all countries except Denmark. The annual number of orthotopic bladder substitutions per institution was calculated as three to four (range of medians for each country) and the number of continent cutaneous diversions as two to seven. Open urethral procedures were performed more frequently in Sweden than in the other countries. Surgery for penile curvature and implantation of three-component prostheses for erectile dysfunction was more commonly performed in Denmark and Iceland compared to Sweden.

[Voiding problems and urinary tract complications in adolescents and adults with myelomeningocele]. / Vannlatingsproblemer og urinveiskomplikasjoner hos ungdom og voksne med myelomeningocele.

BACKGROUND: The aim of this study was to describe voiding dysfunction and urinary tract complications in a population above 16 years of age with myelomeningocele. MATERIAL AND METHODS: 51 persons were included in the study. Data were obtained by questionnaires, ultrasound and glomerular filtration rate; in those with intact urinary bladder, by cystometry and videocystography. RESULTS: 30 out of 33 persons with intact urinary bladder were incontinent. Those with daily incontinence described this as a major problem. Cystometry concluded with normal detrusor contractions in three, detrusor hyperreflexia in five, and a detrusor hyporeflexia in 25 persons. Three out of 30 had vesicoureteral reflux. Ultrasound showed mild hydronephrosis and/or scarring in three persons. Average glomerular filtration rate was 86% (50-131%). 11 had Bricker diversion and seven continent reservoirs. 15 out of 18 persons with urinary diversion were satisfied with this solution. In persons with urinary diversion, the average glomerular filtration rate was 78% (44-109%). Ultrasound showed hydronephrosis and/or scarring in seven out of 16. Overall, urinary tract infections last year were reported by 56%, and pyelonephritis was more common in those with urinary diversion. INTERPRETATION: Incontinence is a common problem in adults with myelomeningocele. About one third had upper urinary tract changes, but none had renal failure.

[Follow up of urinary tract problems in adults with myelomeningocele]. / Oppfølging av urinveisproblemer hos voksne med myelomeningocele.

BACKGROUND: More than 90% of persons with myelomeningocele have a neurogenic bladder disturbance with incontinence and risk of upper urinary tract deterioration. Both aspects need to be considered when planning treatment and follow-up. MATERIAL AND METHODS: The study is based on review of articles and clinical experience. RESULTS: A thorough examination of the patient's voiding methods and incontinence is necessary. Examinations for renal function, reflux and hydroureteronephrosis as well as cystometry should also be carried out. The results of such examinations, together with an assessment of the patient's motor and cognitive function, as well as motivation, will provide a basis for further treatment and follow-up. We suggest a flow-chart for treatment and follow-up of persons above 16 years of age with myelomeningocele. INTERPRETATION: Patients with myelomeningocele should have a thorough examination and an individual plan for treatment and follow-up of their urinary tract dysfunction. Depending on the pathological findings, routine follow-up should be in done intervals from six months to five years.

A fundamental aspect of supportive care delivery: the nurse's opportunity to shape the caring encounter.

Recent advances in supportive care therapies have significantly contributed to the likelihood that a child diagnosed with cancer will be cured. Quantitatively recognized supportive care practices, such as central line care management, pain control, nutritional support, and blood transfusions are typically identified as the major contributors to these improved rates of survival. However, just as important are the more qualitative aspects of the nursing care delivery process. As a fundamental part of nursing's caring role, the encounter with patients and families offers unique opportunities for nurses to participate in the creation of a healing environment. Gaining understanding and skill at balancing professional relationships, while also knowing the risks of caring, are critical components of providing excellent supportive care to pediatric oncology patients and their families. The professional caring encounter is a complex and multidimensional process. Strategies for developing optimal communication and healthy professional caring relationships within the pediatric oncology setting are discussed.

Quantification of DNA binding to cell-surfaces by flow cytometry.

DNA binding to cell-surfaces has been documented in several studies. The interaction of DNA with cells has been shown to have therapeutic potential as a non-viral form of gene delivery and DNA vaccination. Recently, bacterial DNA binding and internalization has been demonstrated in some cells to trigger secretion of cytokines and cell activation. Previous studies to quantify DNA binding to cells have used radiolabeled DNA. Here we report a non-radioactive assay for quantification of cell-surface DNA binding based on the isoparametric analysis of flow cytometric data as described by Chatelier et al., Embo J., 5 (1986) 1181. This assay has the advantage over previously used procedures in not employing radioactive material and being able to discriminate viable from non-viable cells that bind DNA. With the importance of understanding the interaction of DNA with cells, this assay may have application for the identification and characterization of reagents designed to either enhance or inhibit DNA binding to cells.

Novel flow cytometry compensation standards: internally stained fluorescent microspheres with matched emission spectra and long-term stability.

In flow cytometry, the emission spectral overlap of fluorescein and R-phycoerythrin is usually corrected by electronic color compensation using microspheres surface labeled with the same fluorochromes. However, the inherent chemical instability of these fluorochromes may cause inaccurate compensation. To overcome these problems, Compen-Flow beads, a new type of compensation standards were developed. The CompenFlow beads are a set of 6.0-microm-diameter polystyrene microspheres that are internally stained with selected BODIPY dye combinations. When excited by the 488-nm argon laser line, these beads show a nearly perfect emission spectral match to fluorescein-stained, R-phycoerythrin-stained and unstained lymphocytes, respectively. Moreover, since the dye molecules are oil soluble, they are contained inside the microsphere matrix instead of merely on the surface; thus, the molecules are shielded from environmental factors that could affect an exposed fluorochrome. Our results show a stable fluorescence spectral profile and constant intensity for at least 2 years stored either refrigerated or at room temperature.