Secondary amputation after lower extremity free-flap reconstruction.

BACKGROUND: Microsurgical free-tissue transfer is often the definitive reconstructive option for lower extremity limb salvage. Despite an initial successful free-flap reconstruction, some patients ultimately undergo lower extremity amputation. The indications for secondary amputation include non- or malunion, infection, hardware failure, or chronic pain. This study aimed to identify the etiology and outcome of secondary amputation after lower extremity free-flap reconstruction. METHODS: A retrospective cohort study was performed including patients who underwent lower extremity free-flap reconstruction from January 2002 to December 2020. Patients who underwent secondary amputation were identified. A survey based on the PROMIS® Pain Interference Scale and activities of daily living (ADLs) was then conducted to assess patient-reported outcomes. Fifteen (52%) patients who underwent amputation responded to the survey, with a median follow-up time of 4.4 years. RESULTS: Of 410 patients who underwent lower extremity free-flap reconstruction, 40 (9.8%) patients underwent subsequent amputation. Of these, 10 patients had failed free-flap reconstruction and 30 patients had secondary amputation after an initially successful soft tissue coverage. The most common etiology for secondary amputation was infection (68%, n = 27). Eighty percent (n = 12) of survey respondents were able to use a prosthetic limb and ambulate. CONCLUSIONS: The most common etiology of secondary amputation was infection. Most patients who ultimately underwent amputation were able to ambulate with a prosthetic, but the majority of patients reported chronic pain. This study could be used to guide potential free-flap candidates regarding the risks and outcomes of lower extremity free-flap reconstruction.

The relationships between early lactation energy status indicators and endocrine fertility traits in dairy cows.

The relationships between dairy cow milk-based energy status (ES) indicators and fertility traits were studied during periods 8 to 21, 22 to 35, 36 to 49, and 50 to 63 d in milk. Commencement of luteal activity (C-LA) and interval from calving to the first heat (CFH), based on frequent measurements of progesterone by the management tool Herd Navigator (DeLaval), were used as fertility traits. Energy status indicator traits were milk ß-hydroxybutyrate (BHB) concentration provided by Herd Navigator and milk fat:protein ratio, concentration of C18:1 cis-9, the ratio of fatty acids (FA) C18:1 cis-9 and C10:0 in test-day milk samples, and predicted plasma concentration of nonesterified fatty acids (NEFA) on test days. Plasma NEFA predictions were based either directly on milk mid-infrared spectra (MIR) or on milk fatty acids based on MIR spectra (NEFAmir and NEFAfa, respectively). The average (standard deviation) C-LA was 39.3 (±16.6) days, and the average CFH was 50.7 (±17.2) days. The correlations between fertility traits and ES indicators tended to be higher for multiparous (r < 0.28) than for primiparous (r < 0.16) cows. All correlations were lower in the last period than in the other periods. In period 1, correlations of C-LA with NEFAfa and BHB, respectively, were 0.15 and 0.14 for primiparous and 0.26 and 0.22 for multiparous cows. The associations between fertility traits and ES indicators indicated that negative ES during the first weeks postpartum may delay the onset of luteal activity. Milk FPR was not as good an indicator for cow ES as other indicators. According to these findings, predictions of plasma NEFA and milk FA based on milk MIR spectra of routine test-day samples and the frequent measurement of milk BHB by Herd Navigator gave equally good predictions of cow ES during the first weeks of lactation. Our results indicate that routinely measured milk traits can be used for ES evaluation in early lactation.

Rhinopharynx irrigations and mouthwash with dissolved mupirocin in treatment of MRSA throat colonization - proof-of-concept study.

BACKGROUND: To prevent transmission of, and infection with, meticillin-resistant Staphylococcus aureus (MRSA), eradication treatment of colonized individuals is recommended. Throat colonization is a well-known risk factor for eradication failure. Staphylococcus aureus throat colonization is associated with colonization of the rhinopharynx, but in the currently recommended Danish MRSA eradication strategies, rhinopharynx colonization is not directly targeted. Rhinopharynx colonization could therefore be an important risk factor for prolonged MRSA throat carriage. AIM: To determine whether irrigation and wash of the rhinopharynx and mouth with dissolved mupirocin is a feasible and potentially efficacious supplementary strategy against treatment-resistant MRSA throat carriage. METHODS: The patient study was an open, non-blinded, trial including 20 treatment-resistant MRSA throat carriers. In the study, the patients received a supplementary treatment besides the standard treatment according to the Danish MRSA eradication strategy. The supplementary treatment consisted of rhinopharyngeal irrigation and mouth-gurgling twice a day for 14 days with a mupirocin ointment (22 g 2% ointment per litre of isotonic sterile saline solution) in a 37°C solution. FINDINGS: Eighteen patients (90%) complied with the treatment protocol and none ex-perienced any major adverse events. Out of the 18 patients who finished the study per protocol, 15 (83%) and seven (39%) patients had negative MRSA sampling results one and six months after end of treatment, respectively. CONCLUSION: This study demonstrates the feasibility and clinical potential of also targeting the rhinopharynx and oropharynx in non-systemic throat MRSA eradication strategies.

Genetic parameters for reproductive losses estimated from in-line milk progesterone profiles in Swedish dairy cattle.

This study assessed the extent of reproductive losses and associated genetic parameters in dairy cattle, using in-line milk progesterone records for 14 Swedish herds collected by DeLaval's Herd Navigator. A total of 330,071 progesterone samples were linked to 10,219 inseminations (AI) from 5,238 lactations in 1,457 Swedish Red and 1,847 Swedish Holstein cows. Pregnancy loss traits were defined as early embryonic loss (1-24 d after AI), late embryonic loss (25-41 d after AI), fetal loss (42 d after AI until calving), and total pregnancy loss (from d 1 after AI until calving). The following classical fertility traits were also analyzed: interval from calving to first service, interval from calving to last service, interval between first and last service, calving interval, and number of inseminations per service period. Least squares means with standard error (LSM ± SE), heritabilities, and genetic correlations were estimated in a mixed linear model. Fixed effects included breed, parity (1, 2, ≥3), estrus cycle number when the AI took place, and a linear regression on 305-d milk yield. Herd by year and season of AI, cow, and permanent environmental effect were considered random effects. Extensive (approximately 45%) early embryonic loss was found, but with no difference between the breeds. Swedish Red was superior to Swedish Holstein in the remaining pregnancy loss traits with, respectively: late embryonic loss of 6.1 ± 1.2% compared with 13.3 ± 1.1%, fetal loss of 7.0 ± 1.2% compared with 12.3 ± 1.2%, and total pregnancy loss of 54.4 ± 1.4% compared with 60.6 ± 1.4%. Swedish Red also had shorter calving to first service and calving to last service than Swedish Holstein. Estimated heritability was 0.03, 0.06, and 0.02 for early embryonic, late embryonic, and total pregnancy loss, respectively. Milk yield was moderately genetically correlated with both early and late embryonic loss (0.52 and 0.39, respectively). The pregnancy loss traits were also correlated with several classical fertility traits (-0.46 to 0.92). In conclusion, Swedish Red cows had lower reproductive loss during late embryonic stage, fetal stage, and in total, and better fertility than Swedish Holstein cows. The heritability estimates for pregnancy loss traits were of the same order of magnitude as previously reported for classical fertility traits. These findings could be valuable in work to determine genetic variation in reproductive loss and its potential usefulness as an alternative fertility trait to be considered in genetic or genomic evaluations.

Genetic parameters of endocrine fertility traits based on in-line milk progesterone profiles in Swedish Red and Holstein dairy cows.

Evaluating fertility traits based on endocrine progesterone profiles is becoming a promising option to improve dairy cow fertility. Several studies have been conducted on endocrine fertility traits, mainly in the Holstein breed. In this study, focusing also on the Swedish Red (SR) breed, genetic parameters were estimated for classical and endocrine fertility traits, the latter based on in-line milk progesterone records obtained for 14 Swedish herds using DeLaval Herd Navigator (DeLaval International, Tumba, Sweden). A total of 210,403 observations from 3,437 lactations of 1,107 SR and 1,538 Holstein cows were used. Mixed linear animal models were used for estimation of genetic parameters. Least squares means analysis showed that Holstein cows had a 2.5-d-shorter interval from calving to commencement of luteal activity (C-LA) and longer length of first inter-ovulatory interval (IOI) than SR cows. The highest mean interval for C-LA, IOI, and first luteal phase length (LPL) was observed in the fourth parity. The incidence of short (<18 d), normal, (18-24 d), and long (>24 d) IOI was 29.3, 40.7, and 30%, respectively. Genetic analysis indicated moderate heritability (h2) for C-LA (h2 = 0.24), luteal activity during the first 60 d in milk (LA60, h2 = 0.15), proportion of samples with luteal activity (PLA, h2 = 0.13), and calving to first heat (CFH, h2 = 0.18), and low heritability estimates for LPL (h2 = 0.08) and IOI (h2 = 0.03) in the combined data set for both breeds. Similar heritability estimates were obtained for each breed separately except for IOI and LPL in SR cows, for which heritability was estimated to be zero. Swedish Red cows had 0.01 to 0.06 higher heritability estimates for C-LA, LA60, and PLA than did Holstein cows. Calving interval had moderate heritability among the classical traits for Holstein and the combined data set, but h2 was zero for SR. Commencement of luteal activity had a strong genetic correlation with LA60 (mean ± SE; -0.88 ± 0.06), PLA (-0.72 ± 0.11), and CFH (0.90 ± 0.04). Similarly, CFH had a strong genetic correlation with IOI (0.98 ± 0.20). Number of inseminations per series showed a weak genetic correlation with all endocrine traits except IOI. Overall, endocrine traits had higher heritability estimates than classical traits in both breeds, and may have a better potential to explain the actual reproductive status of dairy cows than classical traits. This might favor inclusion of some endocrine fertility traits-especially those related to commencement of luteal activity-as selection criteria and breeding goal traits if recording becomes more common in herds. Further studies on genetic and genomic evaluations for endocrine fertility traits may help to provide firm conclusions. A prerequisite is that the data from automatic devices be made available to recording and breeding organizations in the future and included in a central database.

Danish experience of meticillin-resistant Staphylococcus aureus eradication with emphasis on nose-throat colonization and supplementary systemic antibiotic treatment.

This study aimed to evaluate the Danish Board of Health's guidance for treating the carriage of meticillin-resistant Staphylococcus aureus (MRSA), focusing on nose-throat carriage and use of supplementary systemic antibiotics. The results of MRSA eradication treatment among 358 patients were analysed, focusing on those with nose (N=58) or throat (N=183) MRSA colonization. The Danish guidance for MRSA treatment was found to be more successful in patients with nose colonization (66%) compared with throat colonization (41%), despite the fact that the cumulative eradication rates were equal after three treatment cycles (71% vs 73%). This study found that supplementation of colonization treatment with systemic antibiotics does not have a positive effect.

Genetic parameters for endocrine and traditional fertility traits, hyperketonemia and milk yield in dairy cattle.

High-yielding cows may suffer from negative energy balance during early lactation, which can lead to ketosis and delayed ability of returning to cyclicity after calving. Fast recovery after calving is essential when breeding for improved fertility. Traditionally used fertility traits, such as the interval from calving to first insemination (CFI), have low heritabilities and are highly influenced by management decisions. Herd Navigator™ management program samples and analyses milk progesterone and ß-hydroxybutyrate (BHB) automatically during milking. In this study, the genetic parameters of endocrine fertility traits (measured from milk progesterone) and hyperketonemia (measured from milk BHB) in early lactation were evaluated and compared with traditional fertility traits (CFI, interval from calving to the last insemination and interval from first to last insemination) and the milk yield in red dairy cattle herds in Finland. Data included observations from 14 farms from 2014 to 2017. Data were analyzed with linear animal models using DMU software and analyses were done for first parity cows. Heritability estimates for traditional fertility traits were low and varied between 0.03 and 0.07. Estimated heritabilities for endocrine fertility traits (interval from calving to the first heat (CFH) and commencement of luteal activity (C-LA)) were higher than for traditional fertility traits (0.19 to 0.33). Five slightly different hyperketonemia traits divided into two or three classes were studied. Linear model heritability estimates for hyperketonemia traits were low, however, when the threshold model was used for binary traits the estimates became slightly higher (0.07 to 0.15). Genetic correlation between CFH and C-LA for first parity cows was high (0.97) as expected since traits are quite similar. Moderate genetic correlations (0.47 to 0.52) were found between the endocrine fertility traits and early lactation milk yield. Results suggest that the data on endocrine fertility traits measured by automatic systems is a promising tool for improving fertility, specifically when more data is available. For hyperketonemia traits, dividing values into three classes instead of two seemed to work better. Based on the current study and previous studies, where higher heritabilities have been found for milk BHB traits than for clinical ketosis, milk BHB traits are a promising indicator trait for resistance to ketosis and should be studied more. It is important that this kind of data from automatic devices is made available to recording and breeding organizations in the future.

Characteristics of macrolide responders in persistent post-surgical rhinosinusitis.

INTRODUCTION: The anti-inflammatory effects of long term low dose macrolide therapy have shown benefit in the management of diffuse panbronchiolitis. Dramatic responses to macrolide in the upper airway are seen but our understanding of the patient phenotype predisposing to macrolide response in chronic rhinosinusitis (CRS) is poor. METHODS: A case control study was performed in a tertiary level rhinology practice of consecutive chronic rhinosinusitis patients placed on a 3-month low dose macrolide therapy after failing at least 3 months of corticosteroid irrigation therapy post-endoscopic sinus surgery. Patients were defined as a macrolide responder when having near normal endoscopy after a 3-month period of clarithromycin treatment. Patient characteristics of smoking, asthma, atopy status, revision surgery, symptom severity (SNOT-22) along with biomarkers from serum and tissue histopathology results were compared between groups. RESULTS: Of twenty-eight consecutive macrolide treated patients, 19 responders were compared to 9 non-responders. The groups were similar in age, female gender, non-smoking, asthma, and atopy. Macrolide response was associated with a lack of tissue eosinophilia (more than 10/HPF) and lower serum eosinophilia. Neutrophil expression was similar in tissue and serum. Squamous metaplasia was overexpressed in non-responders. CONCLUSION: Low tissue and serum eosinophilia, and absence of tissue squamous metaplasia may predict a CRS phenotype suitable to a trial of long-term macrolide therapy when surgery and topical therapy has failed.

Estimating probability of insemination success using milk progesterone measurements.

The aim of this study was to quantify the effects of progesterone profile features and other cow-level factors on insemination success to provide a real-time predictor equation of probability of insemination success. Progesterone profiles from 26 dairy herds were analyzed and the effects of profile features (progesterone slope, cycle length, and cycle height) and cow traits (milk yield, parity, insemination during the previous estrus) on likelihood of artificial insemination success were estimated. The equation was fitted on a training data set containing data from 16 herds (6,246 estrous cycles from 3,404 lactations). The equation was tested on a testing data set containing data from 10 herds (8,105 estrous cycles from 3,038 lactations). Predictors were selected to be implemented in the final equation if adding them to a base model correcting for timing of insemination and parity decreased the overall likelihood distance of the model. Selected variables (cycle length, milk yield, cycle height, and insemination during the previous estrus) were used to build the final model using a stepwise approach. Predictors were added 1 by 1 in different order, and the model that had the smallest likelihood distance was selected. The final equation included the variables timing of insemination, parity, milk yield, cycle length, cycle height, and insemination during the previous estrus, respectively. The final model was applied to the testing data set and area under the curve (AUC) was calculated. On the testing data set, the final model had an AUC of 58%. When the farm effect was taken into account, the AUC increased to 63%. This equation can be implemented on farms that monitor progesterone and can support the farmer in deciding when to inseminate a cow. This can be the first step in moving the focus away from the current paradigm associated with poorer estrus detection, where each detected estrus is automatically inseminated, to near perfect estrus detection, where the question is which estrous cycle is worth inseminating?

Collagen matrix as an inlay in endoscopic skull base reconstruction.

BACKGROUND: Multi-layer reconstruction has become standard in endoscopic skull base surgery. The inlay component used can vary among autografts, allografts, xenografts and synthetics, primarily based on surgeon preference. The short- and long-term outcomes of collagen matrix in skull base reconstruction are described. METHODS: A case series of patients who underwent endoscopic skull base reconstruction with collagen matrix inlay were assessed. Immediate peri-operative outcomes (cerebrospinal fluid leak, meningitis, ventriculitis, intracranial bleeding, epistaxis, seizures) and delayed complications (delayed healing, meningoencephalocele, prolapse of reconstruction, delayed cerebrospinal fluid leak, ascending meningitis) were examined. RESULTS: Of 120 patients (51.0 ± 17.5 years, 41.7 per cent female), peri-operative complications totalled 12.7 per cent (cerebrospinal fluid leak, 3.3 per cent; meningitis, 3.3 per cent; other intracranial infections, 2.5 per cent; intracranial bleeding, 1.7 per cent; epistaxis, 1.7 per cent; and seizures, 0 per cent). Delayed complications did not occur in any patients. CONCLUSION: Collagen matrix is an effective inlay material. It provides robust long-term separation between sinus and cranial cavities, and avoids donor site morbidity, but carries additional cost.

Airflow and symptom outcomes between allergic and non-allergic rhinitis patients from turbinoplasty.

BACKGROUND: Inferior turbinate procedures are applied to relieve medically refractory nasal obstruction. However, the nature of congestion differs between allergic(AR) and non-allergic rhinitis(NAR). This study compares surgical outcomes between AR and NAR patients. METHODOLOGY: A case-control study of patients undergoing turbinate with or without septoplasty surgery for nasal obstruction was performed. Patient reported outcomes were: nasal obstruction, global nasal function(GNF), and sino-nasal outcome test(SNOT-22) with rhinitis, facial symptom, sleep and psychological sub-scores. Nasal peak inspiratory flow(NPIF) assessed nasal airflow. Measurements were obtained preoperatively and 3 months postoperatively. RESULTS: 190 patients were assessed. AR had worse obstruction and worse GNF. All outcomes improved post-surgery; nasal obstruction, GNF, SNOT-22, rhinitis-symptoms, facial-symptoms, sleep-function, psychological-function and NPIF. GNF improvement was greater in AR. NPIF improvement was similar between groups. CONCLUSIONS: Both AR and NAR patients gained benefit from surgery to relieve nasal obstruction. AR patients demonstrate greater improvement in GNF score but allergy management may contribute to this.

Periostin as a marker of mucosal remodelling in chronic rhinosinusitis.

BACKGROUND: Although extracellular matrix (ECM) proteins are associated with irreversible lower airway changes, the relationship with upper airway remodelling which occurs during chronic rhinosinusitis (CRS) is poorly understood. This study assessed the expression of ECM proteins periostin, fibulin-1, fibronectin and collagenIV in nasal mucosa of patients with and without histologic features of remodelling. METHODS: A cross-sectional study of sinonasal mucosal biopsies taken from patients, undergoing surgery for CRS was performed, where patients were grouped according to remodelling, defined by basement membrane thickening (BMT over 7.5 micrometer) and subepithelial fibrosis. An overall view and three random fields of immunostained tissue sections that included epithelium, basement membrane and submucosa, were imaged using Zeiss Zen software. The area and intensity of positive staining were scored by two blinded observers, using a 12-point ordinal scale of weak to strong. RESULTS: 65 patients (47.6 +/- 13.4years, 44.6% female) were assessed. Patients were grouped as controls 26.2%, BMT/no fibrosis 38.5% or BMT and fibrosis 33.8%. Stronger grade of periostin expression was associated with remodelling changes and tissue eosinophilia over 10/HPF. Fibulin-1, fibronectin and collagenIV did not differ. CONCLUSION: Periostin expression was associated with the presence of BMT, fibrosis and tissue eosinophilia and may identify patients undergoing remodelling changes.

Association of gastro-oesophageal reflux and chronic rhinosinusitis: systematic review and meta-analysis.

INTRODUCTION: Gastro-oesophageal reflux disease (GORD) has been implicated in the development of chronic rhinosinusitis (CRS). The association of GORD with CRS is systematically assessed from the medical literature. METHODOLOGY: Embase and MEDLINE were searched using a comprehensive strategy limited to English language and Human subjects. Any study with original data on the experimental, diagnostic, treatment or prognostic association of CRS with GORD was included. Studies without a control group, case reports and review articles were excluded. RESULTS: The search returned 958 records, with an additional 10 found from bibliographic lists; this produced 32 studies. The included studies (n=32) consisted of studies reporting pathogenic factors (n=20), epidemiological association (n=8), prognostic interactions (n=3), and a combination of these outcomes (n=1). Potential pathogenic roles for GORD in CRS were supported; CRS subjects had greater prevalence of intranasal Helicobacter pylori and acid reflux than subjects without CRS. CRS is more prevalent in GORD sufferers than those without GORD. Evidence is conflicting for GORD as a factor in CRS treatment failure. CONCLUSION: The results support a significant association of GORD with CRS. Physicians should be cognizant of the potential for acid and non-acid reflux as a driving factor in CRS.

The impact of bitter taste receptor genetics on culturable bacteria in chronic rhinosinusitis.

BACKGROUND: Extra-oral bitter taste receptors have been associated with innate bacterial defence mechanisms. Genetic variation in T2R38 functionality has been shown to be associated with susceptibility to upper respiratory tract infections and chronic rhinosinusitis (CRS). We sought to independently assess the influence of bitter taste receptor genotype on the presence of culturable bacteria in the sinuses. METHODOLOGY: A cross-sectional analysis of patients with CRS undergoing surgery was performed. Middle meatal nasal swabs were sent for microbiological evaluation at the time of the procedure. Mucosal biopsies were taken and sent for bitter taste receptor genotype analysis. Sequencing of 3 polymorphisms in the TAS2R38 gene was performed to identify genotypes as super-tasters (PAV/PAV), non-tasters (AVI/AVI) or heterozygous expression (PAV/AVI). The presence of culturable organisms and common pathogens were compared with bitter taste receptor genotypes. RESULTS: 25 patients (age 52.4 +/- 18.28 years, 51% female) were assessed. Super-tasters comprised 16% of the group, 24% were non-tasters and 48% had heterozygous expression. A cultured pathogen was grown in 48% of patients; 32% gram-positive, 20% gram-negative, 28% grew Staphylococcus aureus and 12% Pseudomonas aeruginosa. A non-taster genotype was predictive of colonised pathogens. Tissue eosinophilia (more than 10 HPF) was seen in 48%. CONCLUSION: Even in a small sample of patients with CRS, non-taster T2R38 genotype appears to predict the presence of culturable bacteria colonising the sinus cavity at the time of surgery for their condition. A genetic link to patients more likely to become infected is likely.

Household air pollution from wood burning in two reconstructed houses from the Danish Viking Age.

During 13 winter weeks, an experimental archeology project was undertaken in two Danish reconstructed Viking Age houses with indoor open fireplaces. Volunteers inhabited the houses under living conditions similar to those of the Viking Age, including cooking and heating by wood fire. Carbon monoxide (CO) and particulate matter (PM2.5 ) were measured at varying distances to the fireplace. Near the fireplaces CO (mean) was 16 ppm. PM2.5 (mean) was 3.40 mg/m(3) , however, measured in one house only. The CO:PM mass ratio was found to increase from 6.4 to 22 when increasing the distance to the fire. Two persons carried CO sensors. Average personal exposure was 6.9 ppm, and from this, a personal PM2.5 exposure of 0.41 mg/m(3) was estimated. The levels found here were higher than reported from modern studies conducted in dwellings using biomass for cooking and heating. While this may be due to the Viking house design, the volunteer's lack of training in attending a fire maybe also played a role. Even so, when comparing to today's issues arising from the use of open fires, it must be assumed that also during the Viking Age, the exposure to woodsmoke was a contributing factor to health problems.

Organophosphorus pesticide degradation product in vitro metabolic stability and time-course uptake and elimination in rats following oral and intravenous dosing.

Levels of urinary dialkylphosphates (DAPs) are currently used as a biomarker of human exposure to organophosphorus insecticides (OPs). It is known that OPs degrade on food commodities to DAPs at levels that approach or exceed those of the parent OP. However, little has been reported on the extent of DAP absorption, distribution, metabolism and excretion. The metabolic stability of O,O-dimethylphosphate (DMP) was assessed using pooled human and rat hepatic microsomes. Time-course samples were collected over 2 h and analyzed by LC-MS/MS. It was found that DMP was not metabolized by rat or pooled human hepatic microsomes. Male Sprague-Dawley rats were administered DMP at 20 mg kg(-1) via oral gavage and i.v. injection. Time-course plasma and urine samples were collected and analyzed by LC-MS/MS. DMP oral bioavailability was found to be 107 ± 39% and the amount of orally administered dose recovered in the urine was 30 ± 9.9% by 48 h. The in vitro metabolic stability, high bioavailability and extent of DMP urinary excretion following oral exposure in a rat model suggests that measurement of DMP as a biomarker of OP exposure may lead to overestimation of human exposure.

Evaluation of a radioimmunoassay and establishment of a reference interval for salivary cortisol in healthy subjects in Denmark.

A commercial radioimmunoassay (RA) for salivary cortisol was evaluated using certified reference material in water and spiked to pooled saliva in the range 2.1-89.1 nmol/L. A variance component model for describing the effects of age, body mass index (BMI), diurnal variation, gender, days of sick leave during the past year, and smoking habits was established. Reference intervals for salivary cortisol in 120 healthy individuals performing their routine work were established according to the International Union of Pure and Applied Chemistry (IUPAC) and the International Federation of Clinical Chemistry (IFCC). The method evaluation of the certified reference material in water did not show any bias of the method, i.e. recovery was 97% [CI: 94%; 100.9%]. LOD (detection limit) was 1.59 nmol/L. The ratio between analytical and within-subject variation (CVa/CVi) was 0.14, indicating that the method was adequate for measurement in healthy subjects. Reference intervals were estimated to be from 3.6 to 35.1 nmol/L for samples at the time of awakening (05.27-07.27), 7.6-39.4 nmol/L for peak level in saliva samples collected 20 min after awakening (05.47-07.47), and LOD 10.3 nmol/L for late afternoon samples (17.00-19.00). Reactivity (increase from awakening to 20 min after awakening) was estimated to be 82% [CI: -179; 345%] and recovery (decrease from 20 min after awakening to 18.00) to be 80% [CI: 51; 109%]. Eighteen percent of the subjects showed a decrease in cortisol in saliva from awakening to 20 min after awakening. Salivary cortisol was not affected by age, body mass index, gender, smoking habits or days of sick leave during the past year.

Reference intervals and variation for urinary epinephrine, norepinephrine and cortisol in healthy men and women in Denmark.

Reference intervals for urinary epinephrine, norepinephrine and cortisol in 120 healthy individuals performing their routine work were established according to the International Union of Pure and Applied Chemistry (IUPAC) and the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) for use in the risk assessment of exposure to occupational stress. Reference intervals were established for three different times of the day: in morning samples (05.45-07.15) the limit of detection (LOD) was 2.10 micromol epinephrine/mol creatinine (82 women) and 2.86 micromol epinephrine/mol creatinine (37 men), and the reference interval was 3.6-29.1 micromol norepinephrine/mol creatinine and 2.3-52.8 micromol cortisol/mol creatinine (119 women and men); in afternoon samples (15.30-18.30) the reference interval was 0.64-10.8 micromol epinephrine/mol creatinine (82 women), 1.20-11.2 micromol/epinephrine/mol creatinine (36 men), 11.0-54.1 micromol/ norepinephrine/mol creatinine and LOD was 42.4 micromol cortisol/mol creatinine (117 women and men); in evening samples (21.45-23.45) LOD was 8.66 micromol epinephrine/mol creatinine (81 women) and 7.99 micromol/epinephrine/mol creatinine (36 men), the reference interval was 11.0-54.1 micromol norepinephrine/mol creatinine, and LOD was 42.4 micromol cortisol/mol creatinine (117 women and men). A variance component model for describing the effects of age, body mass index (BMI), diurnal variation, gender, days of sick leave during past year and smoking habits was established. Women showed a higher morning value but excreted lower amounts of epinephrine during the day as compared to men. No gender differences could be demonstrated for the excretion of norepinephrine and cortisol. Excretion of epinephrine and norepinephrine increased with smoking and decreased with increased BMI. No effects were observed in the excretion of cortisol.

Seasonal and biological variation of urinary epinephrine, norepinephrine, and cortisol in healthy women.

BACKGROUND: There is a significant circadian and seasonal periodicity in various endocrine functions. The present study describes the within-day and seasonal fluctuation for urinary catecholamines and cortisol and estimates the within- (CV(i)) and between-subject (CV(g)) coefficients of variation for healthy women undertaking their routine work. In addition, index of individuality (I(i)) and power calculations were derived. METHODS: Eleven healthy females undertaking their routine life-style at work participated in the study. Each subject collected six samples during 24 h 15 days over a year, giving a total number of 990 samples. Using a random effect analysis of variance, we estimated CV(g) and total within-subject variation (CV(ti)), i.e. combined within-subject and analytical variation, from logarithmically transformed data. Analytical variation was subtracted from CV(ti) to give CV(i). CV(i) was estimated from samples collected monthly during 1 year (CV(iy)), weekly during 1 month (CV(im)), and six to eight times/day (CV(id)). RESULTS: A seasonal variation was demonstrated for excretion of epinephrine, norepinephrine, and cortisol standardized with creatinine. Concentrations of urinary epinephrine were higher during June and July compared to the rest of the year, whereas concentrations of urinary cortisol were higher during December and January compared to the rest of the year. Excretion of norepinephrine was lower during working hours and higher during hours off work for June and July compared to the rest of the year. There was a high within- and between-subject variation, which could not be explained by menstrual cycle, behavioral, emotional, or cognitive stress reactions. CONCLUSIONS: Despite high biological variation a reasonably low sample size, e.g. 10-50 individuals, is adequate for practical applicability, i.e. studying differences above 150%. The present study recommends to include the sampling time in the statistical evaluation of data and to be aware of the changes in diurnal variations over seasons. When single measurements are to be evaluated, reference intervals are recommended.