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.

COVID-19 related chemosensory changes in individuals with self-reported obesity.

BACKGROUND/OBJECTIVES: Self-reported smell loss is a prominent symptom of COVID-19 infection and a potentially useful clinical tool for remote pre-screening of this disease. However, pre-existing chemosensory dysfunction with obesity may diminish the usefulness of self-reported smell loss in this vulnerable group. Here we aim to compare COVID-19 related chemosensory alterations in participants with and without obesity and determine if self-reported smell loss is predictive of lab-based COVID-19 diagnosis in both groups in the context of restrictive clinical data collection. SUBJECTS/METHODS: In this secondary analysis of a cross-sectional global dataset, we compared self-reported chemosensory ability in participants with a respiratory illness reporting a positive (C19+; n = 5156) or a negative (C19-; n = 659) COVID-19 laboratory test outcome, who also self-reported to have obesity (C19+; n = 433, C19-; n = 86) or not. RESULTS: Participants with obesity and without obesity reported a similar decline in smell, taste, and chemesthesis during illness. In C19+ participants with obesity, we observed a greater relative prevalence of non-chemosensory symptoms, including respiratory and GI symptoms. Critically, we found that the model previously proposed also predicts C19+ diagnosis in participants with obesity. CONCLUSIONS: We conclude that COVID-19 respondents with obesity experience a similar self-reported chemosensory loss as those without obesity. In both groups self-reported chemosensory symptoms are similarly predictive of COVID-19 infection, thus highlighting the potential of collecting self-report of symptoms and comorbidities remotely when clinical observations are restrictive.

COVID-19 Related Chemosensory Changes in Individuals with Self-Reported Obesity.

BACKGROUND/OBJECTIVES: Individuals with obesity show alterations in smell and taste abilities. Smell and taste loss are also the most prominent neurological symptoms of COVID-19, yet how chemosensory ability present in individuals with obesity with a positive COVID-19 diagnosis is unknown. SUBJECTS/METHODS: In this secondary analysis of a cross-sectional global dataset, we compared self-reported chemosensory ability in participants with a respiratory illness reporting a positive (C19+; n = 5156) or a negative (C19-; n = 659) COVID-19 laboratory test outcome, who also self-reported to be obese (C19+; n = 433, C19-; n = 86) or non-obese. RESULTS: Compared to the C19- group, C19+ exhibited a greater decline in smell, taste, and chemesthesis during illness, though these symptoms did not differ between participants with obesity and without obesity. In 68% of participants who reported recovery from respiratory illness symptoms (n=3431 C19+ and n= 539 C19-), post-recovery chemosensory perception did not differ in C19+ and C19- diagnosis, and by self-reported obesity. Finally, we found that all chemosensory and other symptoms combined predicted the C19+ diagnosis in participants with obesity with a moderately good estimate (63% accuracy). However, in C19+ participants with obesity, we observed a greater relative prevalence of non-chemosensory symptoms, including respiratory as respiratory and GI symptoms. CONCLUSIONS: We conclude that despite a presumed lower sensitivity to chemosensory stimuli, COVID-19 respondents with obesity experience a similar self-reported chemosensory loss as those without obesity, and in both groups self-reported chemosensory symptoms are similarly predictive of COVID-19.

Flavors prime processing of affectively congruent food words and non-food words.

The aim of this study was to explore the use of oral flavor stimuli in an implicit measure of attitudes; the affective priming paradigm. Unpleasant (cold instant coffee) and pleasant (strawberry lemonade) chemosensory flavor stimuli were used as primes in an affective cross-modal priming paradigm. Target stimuli were food words and non-food words, that were either affectively positive or negative, thus creating affectively congruent and incongruent prime-target pairs. We observed priming for congruent flavor-word pairs, i.e. if prime and target are both positive or both negative, this led to faster evaluation of the target words than for incongruent flavor-word pairs. Furthermore, the size of the priming effect was similar for food and non-food target words, suggesting that the affective priming effect is not augmented by the use of words that are semantically related. These results provide proof of concept of indirectly measuring attitudes to flavors with the affective priming paradigm, which may provide information on attitudes in addition to explicit pleasantness ratings.

Selected trace elements and minerals in cord blood: association with lipids and lipoproteins at birth.

Recent studies have found that some minerals are associated with lipoprotein metabolism, peroxidation and coronary heart disease. The present study was designed to obtain information on the relationship between the concentrations of minerals and trace elements and those of lipids, lipoproteins and apolipoproteins (Apo) in cord blood. Seventy-nine healthy singleton normoweight full-term newborns without foetal distress according to Apgar scores were studied. The quartile distribution of neonates, according to serum concentrations of the different minerals studied, displayed significant differences in lipid values between infants in the lowest and highest quartiles of: magnesium, for HDL-cholesterol, Apo A-I, Apo B and the HDL-cholesterol/Apo A-I ratio; iron, for HDL-cholesterol and the HDL-cholesterol/Apo A-I ratio; copper, for triglycerides and the LDL-cholesterol/Apo B ratio. With the exception of calcium, several significant correlations were found between the minerals and trace elements and the lipids, Apo and lipoproteins tested.

Prevalence of delta F508 cystic fibrosis carriers in The Netherlands: logistic regression on sex, age, region of residence and number of offspring.

An average cystic fibrosis (CF) carrier frequency of 1 in 25 in Europe is cited in numerous reports, although a great variability in estimated prevalences has been found in different European populations. The estimates of these frequencies were based on numbers of CF patients before identification of the gene in 1989. Here we report the results of a study to determine the carrier frequency of the delta F508 mutation in The Netherlands by analyzing mouthwashes and matched blood samples from 11654 blood donors all over the country. We analyzed possible relationships between a number of theoretically explanatory variables and the delta F508 carrier frequency by means of univariate and multivariate logistic regression. These variables were: distance of the blood banks from the northeastern part of the country (distance); whether the blood donors knew that we were looking for a CF mutation; sex and age of the donor; and number of children of the donor (family size). We detected a delta F508 carrier frequency of 1 in 42 (95% CI 1/37-1/47) in The Netherlands. If we assume that the relative frequency of the delta F508 mutation among carriers and patients is comparable in The Netherlands, this gives an estimated overall CF carrier frequency of 1 in 32 (95% CI 1/28-1/36), significantly less than 1 in 25. The univariate logistic regression analysis of the effects of the explanatory variables on the carrier frequency revealed no significant relationships, except for an increase in carrier frequency with increasing distance from the northeastern region. In the multivariate analysis with all five independent variables, distance, age and family size were significantly related to the carrier frequency, but sex and CF information were not. There was a significant interaction between age and family size. In our final model, distance, age and family size were positively related to the carrier frequency, while the interaction of age with family size showed a negative relation. These results confirm that there is a gradient in gene frequency with low frequencies in the northeastern part of the country and high frequencies in the southern part. They also suggest a relation of age and family size with carrier frequency. This relation, however, is too complex to be explained by heterozygote advantage.

Growth characteristics in laboratory animals fed zinc-deficient, copper-deficient, of histidine-supplemented diets.

The effects of growth in male Wistar rats and female Swiss Random mice were studied during dietary zinc (Zn) deficiency, copper (Cu) deficiency, and during the feeding of a histidine (His) supplement Growth was analyzed by comparing the characteristics of the decreasing exponential growth curve plotted for the experimental period. When the animals were pair-fed the experimental diets, the growth pattern in the animals remained unaltered. The growth rate decreased during Zn deficiency by a factor of 0.64 over a period of 10 d (male young adult rats) and by a factor of 0.76 over a period of 28 d (female weaning mice). On the other hand, a supplement of His increased the growth rate by a factor of 1.11 (in the mice). The effect of Cu deficiency on the growth rate was not statistically significant (in the rats). However, Cu deficiency causes effects in the Zn status that may over-compensate minor growth retardation during Cu deficiency. The effect of the His supplement is explained by its having an effect on the Zn-absorption (His enhancing Zn transport over the gut) and by a stimulating effect of this amino acid on the thickness of the growth plate in bone.

Validation of the determination of deltaF508 mutations of the cystic fibrosis gene in over 11 000 mouthwashes.

Mouthwashes can be used as a DNA resource for mutation detection and, because collection and DNA isolation is simple and cheap, they could in particular, be used for large numbers of samples. To determine the failure rate (the proportion of mouth samples in which no PCR product was obtained) and the specificity of buccal epithelial cell mutation detection in large numbers of samples, we collected mouthwashes and blood samples from 11 413 blood donors and tested the mouthwashes for the deltaF508 mutation, which has an estimated frequency Of 75% among cystic fibrosis chromosomes in The Netherlands. Blood samples were tested for the deltaF508 mutations only if the mutation was identified in the mouthwash or in the case of a failure to obtain PCR products. The sensitivity of the test was determined in mouthwashes of 75 deltaF508 carriers known from earlier family studies. These samples were offered blindly between the mouthwashes of the blood donors. Both specificity and sensitivity of the mouthwash procedure were 100%. The overall failure rate was 5.6%. This large figure was caused mainly by insufficient rinsing of the mouth in one particular blood bank. Exclusion of the results of this blood bank reduced the failure rate to 1.8%. Our results also confirmed that for a large number of samples the mouthwash procedure is suitable for mutation detection and, with proper instructions, can be used in community screening.

Dietary subacute zinc deficiency and potassium metabolism.

In a controlled animal experiment the effects of dietary subacute Zn deficiency on growth, Zn concentration, and tissue 42-K distribution were studied. Growth retardation caused lower body weight because both skeletal and heart muscle showed a reduction in cell mass. Zn concentrations were reduced in most tissues, however, they remained unaltered in heart muscle. 42-K activity increased in skeletal muscle and pancreas. We hypothesize the latter reflects the organs rate of metabolism, inducing the exocrine pancreas to increase Zn absorption; in skeletal muscle it may induce also alterations in cell potentiation, causing restless behavior. As suggested by the calculated specific K activity (Bq/mol), the K uptake was highest in liver and bone, high in pancreas and skeletal muscle and low in heart muscle. The latter suggests K retention in heart muscle. Specific activity in plasma and jejunum remained unaltered: K status and absorption seem unaffected. Zn deficiency causes different 42-K activities in the various tissues, that respond by alterations in K metabolism without the induction of K deficiency.

Effects of neonatal exposure to clonidine on basal and activated central noradrenaline metabolism and in vivo overflow.

The occurrence of persistent effects of chronic neonatal exposure to the alpha 2-adrenoceptor agonist clonidine was investigated by determination of tissue concentrations of monoamines and metabolites and in vivo overflow of noradrenaline and its metabolites, in various rat brain regions during adulthood. Rat pups were treated with clonidine from postnatal day 10-20 and all measurements were carried out between postnatal day 40 and 58. Tissue concentrations of monoamines and metabolites of the early clonidine-treated rats did not differ significantly from the control group. A challenge with yohimbine did not reveal altered responses of monoaminergic systems, except for the failure of an increased serotonergic activity in the medulla pons. In vivo microdialysis measurements revealed an elevated basal extracellular noradrenaline level in amygdala, but not in frontal cortex and hippocampus. Pharmacological challenge in vivo with idazoxan did not reveal differences between clonidine- and saline-exposed rats. These results confirm previous findings that continuous activation of alpha 2-adrenoceptors during a particular period of rat brain development may result in long-lasting but small changes in monoaminergic activity. However, these alterations are not very consistent and may depend on the parameter chosen to reflect monoaminergic activity and are not revealed more clearly by activating (challenging) the noradrenaline system with alpha2-adrenoceptor antagonists.

Increased alpha 2-adrenoceptor mediated regulation of adult rat brain noradrenaline overflow after chronic neonatal exposure to propranolol; a microdialysis study.

Direct and persistent effects of chronic neonatal administration of the beta-adrenoceptor antagonist propranolol on brain noradrenergic activity were investigated by measuring tissue concentrations of noradrenaline and its metabolites and in vivo overflow of noradrenaline during adulthood. Rat pups were chronically treated with propranolol from postnatal day 1 to day 10. Determination of monoamine metabolism after the last injection showed an increase in noradrenaline metabolism in frontal cortex, limbic system and hippocampus of propranolol-exposed rats, but 47 days after this last injection it was apparent that these effects were not long-lasting. Moreover, basal noradrenaline overflow in vivo in the hippocampus of 40-55 day-old propranolol-exposed rats did not differ from that in controls. However, the regulation of noradrenaline release seemed to have been altered, since a pharmacological challenge with the alpha 2-adrenoceptor antagonist idazoxan induced an enhanced increase in the in vivo noradrenaline overflow in propranolol-exposed rats compared to controls. It is suggested that the neonatal beta-blockade induced a supersensitivity of the presynaptic alpha 2-adrenoceptor. The precise mechanism underlying this effect has to be elucidated.

Regional differences in the in vivo regulation of the extracellular levels of noradrenaline and its metabolites in rat brain.

Microdialysis was used to determine extracellular levels of both noradrenaline and its metabolites in several brain regions of rats under basal conditions and in response to drugs selective for the alpha 2-adrenoceptor to study regional differences in the regulation of noradrenaline overflow. Basal overflow of noradrenaline was about 1.3 fmol/min in frontoparietal cortex, amygdala and hippocampus and in the medial prefrontal cortex 2.4 fmol/min was measured, whereas the overflow of the noradrenaline metabolites 3,4-dihydroxyphenylglycol and 3-methoxy-4-hydroxyphenylglycol was 10-fold higher. After correction for recovery and membrane length no regional differences in the basal overflow of noradrenaline (NA) were found. There were, however, regional differences in the drug-induced effects: locally applied moxonidine decreased extracellular noradrenaline stronger in the frontoparietal cortex than in the medial prefrontal cortex. The increase in noradrenaline overflow caused by idazoxan (10(-4) M) was stronger in frontoparietal cortex than in amygdala and hippocampus. The metabolites were also generally decreased by moxonidine and increased by idazoxan, although less markedly. The present study shows that the regulation of noradrenaline overflow by the presynaptic alpha 2-autoreceptor was stronger in cortical regions than in amygdala and hippocampus. In those latter regions the uptake mechanism probably plays a relatively more important role in the regulation of noradrenaline overflow.

The uptake of zinc by erythrocytes under near-physiological conditions.

The in vitro uptake of zinc by erythrocytes was measured under near-physiological conditions, using 65Zn as a radioactive tracer. Because of the presence of serum albumin--a strong zinc ligand--a low concentration of medium free zinc was maintained. Under these conditions a high-affinity carrier for zinc transport over the cell membrane was identified. With human erythrocytes, a Michaelis constant (Km) of 0.2 nM with respect to free medium zinc was measured and a Vmax of 4.5 nmoles Zn transported per h/g dry wt. The Km for medium Zn increases when the size of the internal erythrocytic Zn pool is augmented, whereas Vmax remains virtually unchanged. A model to explain this phenomenon is proposed. It is suggested that this phenomenon could underlie observations, confirmed here, that the in vitro uptake of Zn by animal erythrocytes depends on the Zn status of the animal.

In vivo microdialysis of noradrenaline overflow: effects of alpha-adrenoceptor agonists and antagonists measured by cumulative concentration-response curves.

1. The purpose of the present study was to compare the effects of several alpha-adrenoceptor agonists and antagonists on cerebral cortical overflow of endogenous noradrenaline (NA) in freely moving rats. One or two days after the implantation of transcerebral dialysis tubes in the frontoparietal cortex, extracellular NA levels were monitored on-line with high performance liquid chromatography and electrochemical detection. The drugs were applied locally via the dialysis membrane, and effects on NA overflow were determined in cumulative concentration-response curves. 2. The average basal cortical NA overflow of all experiments was 0.25 pg min-1. The alpha 2-adrenoceptor agonists caused a concentration-dependent decrease in NA levels. UK-14,304 was the most potent and B-HT 933 the least potent agonist. The maximal decrease in NA overflow was to 10-15% of control levels after UK-14,304 or moxonidine, to 30% after clonidine and to 50% after B-HT 933 administration. Continuous activation of the presynaptic alpha 2-adrenoceptor with 10(-6) M UK-14,304 caused a decrease in NA levels to 40-50% of basal levels. This decrease was reached within 1 h and remained stable for the entire 3 h measurement period. The alpha 1-adrenoceptor agonists, phenylephrine and methoxamine, induced an increase in NA levels to 225% and 300%, respectively, at a concentration of 10(-3) M. 3. Local application of alpha 2-adrenoceptor antagonists caused an increase in NA levels, with idazoxan being more potent than piperoxan. Yohimbine did not cause any significant change. 4. All drugs used in these in vivo experiments had in vitro recoveries across the dialysis membrane between 10 and 20%. 5. We conclude that microdialysis with local drug application is suitable for the comparison of the pharmacological effects of drugs with affinity for alpha-adrenoceptors on cortical NA overflow in vivo,provided that the passage across the membrane is equal for the different drugs.

Laboratory assessment of early dietary, subclinical zinc deficiency: a model study on weaning rats.

Male weaning rats were pair-fed a low-zinc diet or a control diet. After 10 d, the animals fed the low-zinc diet showed physiologic signs of deficiency; however, they showed no clinical symptoms. Their estimated whole body zinc was 25 mumol versus 39 mumol for the controls. The 65Zn absorption increased 2-fold and the tissue distribution altered: muscle and erythrocytes contained more, small intestine and liver less activity at 0.5 h postdose. In vitro, the erythrocyte 65Zn uptake rate increased also. The 65Zn uptake experiments required small quantities of erythrocytes. The difference observed between the deficient and control cells was significant and showed little overlap. The increase of the 65Zn uptake from a medium was not affected when the animals underwent endotoxin exposure 24 h before, as was reported to occur in whole blood 65Zn uptake. Therefore, we suggest the in vitro erythrocyte 65Zn uptake, performed in a standardized, near physiologic medium, to detect early, subclinical zinc deficiency.

Dietary vitamin E deficiency and zinc metabolism.

In adult rats, the influence of vitamin E deficiency on zinc metabolism in general and specifically in 15 tissues was studied. After 50 days, we found evidence of vitamin E deficiency and at this time point we injected a tracer amount of 65Zn. During the next 18 days the zinc status was unaffected. The zinc metabolism, however, was altered: the apparent retention increased and the biological half life was prolonged. On day 68, the changes in various tissues varied. Some tissues were affected in zinc concentration (higher in plasma and spleen; lower in cerebrum, fur and tail), others in specific activity (higher in pancreas; lower in cerebellum). The different effects may reflect differences in tissue response on impairement, caused by vitamin E deficiency.

Microdialysis studies on cortical noradrenaline release: basic characteristics, significance of extracellular calcium and massive post-mortem increase.

Extracellular noradrenaline was measured on-line in the cerebral cortex of the freely moving rat by microdialysis coupled to high performance liquid chromatography and electrochemical detection. High potassium concentrations in the perfusion fluid led to a strong increase, whereas tetrodotoxin led to almost undetectable levels of noradrenaline. This shows that noradrenaline in the dialysate was directly derived from active neuronal release. Noradrenaline levels were sensitive to calcium concentrations in the perfusion fluid: from 10% in calcium-free medium to 200% for 10 mM calcium. An overdose of anesthetic caused a 60-fold increase of noradrenaline post-mortem.

Discrimination between low dietary zinc and endotoxin exposure: a model study on weaning rats.

To establish a parameter for zinc status that is independent of the occurrence of infection, we studied the effects of low dietary zinc and endotoxin in weaning rats 21 d after 65Zn intubation. We monitored aspects of zinc status (tissue zinc content, 65Zn distribution, and specific 65Zn activity in tissue) and 65Zn metabolism (absorption, excretion, and biologic half-life), as well as weight gain, feed conversion, and dietary zinc use. The low zinc diet induced classical deficiency with losses of bone zinc, resulting in lower content (7.4 versus 19.6 mumol) and higher spec act (17 versus 8 kBq/mumol). Other tissue-specific and plasma-specific activities were also higher (overall, 20 versus 8 kBq/mumol; plasma, 8 versus 4 kBq/mumol). Endotoxin caused lower total-plasma zinc (0.04 versus 0.05 mumol) but did not affect spec act (4 kBq/mumol); combined endotoxin and low-zinc diet caused low total-plasma zinc (0.01 mumol) and high spec act, as did the low-zinc diet alone (12 kBq/mumol). We conclude that plasma-spec act (or stable isotope enrichment) can serve as an index for nutritional zinc status during recurrent infection.

In vitro exchangeable erythrocytic zinc.

Exchangeable erythrocytic zinc is measured by 65Zn uptake in and release from erythrocytes under standardized and near, physiological conditions: 7.6 microM zinc and 580 microM albumin in the medium. The intracellular exchangeable erythrocytic zinc pool in healthy volunteers amounts to 5 mumol zinc/L packed cells. The half-time of the exchange is 7 h, its activation energy 84 kJ/mol. The effects of the variation in temperature and the concentrations of albumin, as well as the effects of some zinc carriers, cell transport inhibitors, and stress hormones on the 65Zn uptake are measured.

Clinical examination of weanling rats with early zinc deficiency.

In order to gain experience about the detection of adverse effects during a scientific procedure, we carried out a clinical examination of rats with zinc deficiency. In weanling rats fed a zinc-deficient diet (30 mumol zinc/kg) for 10 days, the mean tibial concentration of zinc was reduced by 53% and body weight gain by 73% when compared with rats fed a diet containing an adequate amount of zinc (150 mumol zinc/kg). In a small open field on day 9 of the experiment, the deficient rats more frequently displayed the posture standing upright with elevated heels. On day 10 of the experiment a clinical examination was carried out at random and 'blind' by three independent assessors. Out of 20 variables scored quantitatively on each individual animal, only body size differed between normal and deficient rats. Other classical signs of zinc deficiency, such as alopecia, dermatitis and diarrhoea, were not detected. It is concluded that in this rat model of zinc deficiency, no evidence for extreme discomfort can be demonstrated.