Metabolic engineering of Saccharomyces cerevisiae for the de novo production of psilocybin and related tryptamine derivatives.

Psilocybin is a tryptamine-derived psychoactive alkaloid found mainly in the fungal genus Psilocybe, among others, and is the active ingredient in so-called "magic mushrooms". Although its notoriety originates from its psychotropic properties and popular use as a recreational drug, clinical trials have recently recognized psilocybin as a promising candidate for the treatment of various psychological and neurological afflictions. In this work, we demonstrate the de novo biosynthetic production of psilocybin and related tryptamine derivatives in Saccharomyces cerevisiae by expression of a heterologous biosynthesis pathway sourced from Psilocybe cubensis. Additionally, we achieve improved product titers by supplementing the pathway with a novel cytochrome P450 reductase from P. cubensis. Further rational engineering resulted in a final production strain producing 627 ± 140 mg/L of psilocybin and 580 ± 276 mg/L of the dephosphorylated degradation product psilocin in triplicate controlled fed-batch fermentations in minimal synthetic media. Pathway intermediates baeocystin, nor norbaeocystin as well the dephosphorylated baeocystin degradation product norpsilocin were also detected in strains engineered for psilocybin production. We also demonstrate the biosynthetic production of natural tryptamine derivative aeruginascin as well as the production of a new-to-nature tryptamine derivative N-acetyl-4-hydroxytryptamine. These results lay the foundation for the biotechnological production of psilocybin in a controlled environment for pharmaceutical applications, and provide a starting point for the biosynthetic production of other tryptamine derivatives of therapeutic relevance.

Association of mortality with out-of-hours admission in patients with perforated peptic ulcer.

BACKGROUND: Perforated peptic ulcer is a serious emergency surgical condition. The aim of the present nationwide cohort study was to evaluate the association between mortality and out-of-hours admission in patients surgically treated for perforated peptic ulcer. METHODS: All Danish patients surgically treated for benign gastric or duodenal perforated peptic ulcer in Denmark between September 1, 2011 and August 31, 2013 were included. Patients were identified through The Danish Clinical Register of Emergency Surgery. The association between 90-day mortality and time and day of admission and surgery was assessed by crude and adjusted odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: A total of 726 patients were included. Median age was 69.5 years (range 18.2-101.7), and 569 of the 726 patients (78.4%) had at least one coexisting disease. Adjusted ORs and 95% CIs between 90-day mortality and admission in daytime vs. nighttime and weekday vs. weekend were 1.0 (0.7-1.5) and 1.2 (0.8-1.8), respectively. Adjusted ORs with 95% CI between surgery in daytime vs. nighttime and weekday vs. weekend were 0.9 (0.6-1.3) and 1.2 (0.8-1.8), respectively. Sensitivity analysis was consistent with the primary analysis. The overall 90-day mortality rate was 25.6% (186/726). CONCLUSION: No statistically significant adjusted association between 90-day mortality and out-of-hours admission was found in patients surgically treated for perforated peptic ulcer.

Serum Tg--a sensitive marker of thyroid abnormalities and iodine deficiency in epidemiological studies.

Serum Tg is widely used in the control of thyroid cancer but also in the diagnosis of certain other thyroid diseases. Serum Tg may be useful in the characterization of the iodine status of a population, but little is known about determinants of serum Tg levels. We examined a random selection of 4,649 subjects from 2 regions in Denmark with different iodine status. Thyroid volume and structure were determined with ultrasonography, and thyroid function tests and Tg analysis were performed. The factor with the closest association with serum Tg levels was thyroid volume at ultrasonography (P < 0.001). Also thyroid nodularity (P < 0.001) and iodine excretion (P < 0.001) had close associations to serum Tg, even after adjusting for the influence of the other parameters. Thyroid dysfunction had a less pronounced but still highly significant association with serum Tg (P < 0.001), but no relation was found to serum TSH in general. The association with age seemed to rely on differences in the prevalence of thyroid abnormalities, and men had lower Tg levels than women of the same age. There was a marked difference in serum Tg between the two regions with slightly different iodine excretion also after adjusting for the other factors. In conclusion, serum Tg reflects thyroid abnormalities and thyroid function and is a sensitive marker of iodine deficiency in a population.

Folate intake, lifestyle factors, and homocysteine concentrations in younger and older women.

BACKGROUND: An elevated total homocysteine (tHcy) concentration is considered to be an independent risk factor for cardiovascular diseases and has also been associated with an increased risk of neural tube defects. OBJECTIVE: The objective of this study was to investigate folate intake, folate status, and the association between folate intake, other dietary and lifestyle factors, and tHcy concentrations in young and older women. DESIGN: tHcy concentrations were measured in 290 young women aged 25-30 y and in 288 older women aged 60-65 y. All participants completed questionnaires about factors including lifestyle, health, and use of vitamin supplements. Red blood cell folate was measured in 204 of the participants. A subgroup of 258 participants completed dietary records. RESULTS: Median tHcy was 7.6 micromol/L (range: 6.5-8.9) in the younger women and 9.4 micromol/L (7.7-11.1) in the older women. Folate intake from diet was 283 (224-348) and 268 (210-326) microg/d, respectively, in the 2 age groups. Folic acid intake from supplements (P: < 0.001 for the younger women and P: = 0.026 for the older women) and total folate intake (P: = 0.024 and P: = 0.079) were inversely associated with log tHcy in multiple linear regression analyses. Smoking status, coffee consumption, systolic blood pressure, and body mass index were positively associated and estrogen replacement therapy and tea consumption were inversely associated with log tHcy in some of the models. CONCLUSIONS: According to the criteria used, between 1% and 36% of the women had suboptimal folate intake. Folic acid is a strong predictor of tHcy concentration; however, several dietary and other lifestyle factors seem to be important as well.

High occurrence of thyroid multinodularity and low occurrence of subclinical hypothyroidism among tobacco smokers in a large population study.

Tobacco smoking increases the risk of goitre and Graves' disease, but the association with thyroid nodularity and hypothyroidism has not been settled. We investigated 4649 subjects from the general population with questionnaires, thyroid ultrasonography and blood tests. The results were analysed in multivariate regression models. Tobacco smoking was associated with an increased prevalence of thyroid multinodularity (odds ratio (OR) 1.9; 95% confidence interval (CI) 1.4-2.5), but not with increased prevalence of solitary thyroid nodules. The tendency was for a stronger association in the area with the most pronounced iodine deficiency (P for interaction=0.08). Lower levels of serum TSH were found among tobacco smokers (P<0.001), but this association disappeared when adjustment was made for thyroid nodularity and thyroid Volume. The prevalence of elevated TSH levels was markedly reduced among smokers (OR 0.47; 95% CI 0.33-0.67). No association was found between smoking and hyperthyroidism. The observed associations seem to be explainable by the blocking of iodine uptake and organification in the thyroid by thiocyanate, a degradation product of cyanide in tobacco smoke.

Comparative study of thyroid function and types of thyroid dysfunction in two areas in Denmark with slightly different iodine status.

OBJECTIVE: The pattern of thyroid dysfunction seems to depend on the iodine status of the population. Prevalence of thyroid dysfunction could be a parameter to consider when evaluating iodine deficiency disorders in a population. DESIGN: Comparative cross-sectional investigation in two regions in Denmark with marginally different iodine excretion. METHODS: A random selection of 4649 participants from the Civil Registration System in Denmark in age groups between 18 and 65 years were examined. Thyroid dysfunction was evaluated from blood samples and questionnaires, and compared with results from ultrasonography. RESULTS: Median iodine excretion was 53 microg/l in Aalborg and 68 microg/l in Copenhagen. Previously diagnosed thyroid dysfunction was found with the same prevalence in the regions. Serum TSH was lower in Aalborg than in Copenhagen (P=0. 003) and declined with age in Aalborg, but not in Copenhagen. Not previously diagnosed hyperthyroidism was found with the same overall prevalence in the regions, but in age >40 years hyperthyroidism was more prevalent in Aalborg (1.3 vs 0.5%, P=0.017). Not previously diagnosed hypothyroidism was found more frequently in Aalborg (0.6 vs 0.2%, P=0.03). Hyperthyroidism was more often associated with macronodular thyroid structure at ultrasound in Aalborg and hypothyroidism was more often associated with patchy thyroid structure in Copenhagen. CONCLUSIONS: Significant differences in thyroid dysfunction were found between the regions with a minor difference in iodine excretion. The findings are in agreement with a higher prevalence of thyroid autonomy among the elderly in the most iodine-deficient region.

Thyroid structure and size and two-year follow-up of solitary cold thyroid nodules in an unselected population with borderline iodine deficiency.

OBJECTIVE: Multinodular goitre has been found with a high prevalence in iodine-deficient areas, but less frequently in iodine-replete areas; the iodine intake sufficient to prevent goitre has not been established, however. METHODS: We report data from an ultrasonic investigation of the thyroid glands of 2656 randomly selected subjects aged 41 to 71 years in an area with borderline iodine deficiency. RESULTS: Median iodine concentration in spot urine samples was 70microg/l. Multinodular thyroid structure was found in 23% of the population, increasing in women from 20 to 46% with increasing age, and in men from 7 to 23%. Solitary, scintigraphically cold, thyroid nodules >10mm were found in 2.4% of the population with the same prevalence in the different age and sex groups. Two years of follow-up of these cold nodules revealed no signs of malignancies. Median thyroid volume was 11.0ml. Thyroid enlargement (>18ml for women and >25ml for men) was found among 13. 1% of the women and 6.2% of the men, and the prevalence increased with age. The presence of thyroid nodules was related to positive anti-thyroperoxidase antibody (TPO Ab) titres, whereas thyroid enlargement was associated with iodine excretion <50microg/day. CONCLUSIONS: Thyroid enlargement was associated with low iodine excretion and median thyroid volume was slightly increased compared with iodine-replete areas. Multinodular thyroid structure was found with a high prevalence and was associated with TPO Ab >200kU/l. Cold thyroid nodules were moderately prevalent, with no cases of detected malignancies during 2 years of follow-up.

Simultaneous registration of sleep-stages and bladder activity in enuresis.

A combined monitoring of sleep stages and bladder activity for the evaluation of enuresis is on trial. Polygraphic recordings in 7 patients, comprising the preliminary material, have shown single-event bladder contractions without forewarning or changes in sleep stages. A volume dependency of the enuresis episodes seems so far to be a major mechanism.

Review of thyroid cancer cases among patients with previous benign thyroid disorders.

A previous register linkage study showed an increased risk of thyroid cancer among patients previously discharged from a hospital with a diagnosis of a benign thyroid disorder. In this study, we have reviewed all available medical records, first to validate the earlier result and second to describe the symptomatology of patients with a history of benign thyroid disorder prior to the cancer diagnosis. The previous study identified 189 patients with a benign and subsequent malignant thyroid disorder. Medical records were obtainable for 156 of these patients and were reviewed. For 104 patients, benign and malignant thyroid diseases were metachronous (a clearly separated disease history of the benign and malignant diseases), and for 48 patients synchronous. In 4 cases, thyroid cancer could not be confirmed. Among patients with metachronous thyroid disorders, all major benign thyroid disorders were represented including hot nodules, diffuse and multinodular toxic and nontoxic goiter. Symptoms preceding diagnosis of thyroid cancer included growth of goiter/nodules, globulus, stridor, hoarseness, and metastasis. No major differences were found among patients with metachronous and synchronous benign and malignant thyroid disorder, apart from the fact that all metastases were found among metachronous cases. This study confirmed the conclusion that patients with a previous history of goiter or nodules have an increased risk of thyroid cancer. However, thyroid cancer still occurs too infrequently to warrant screening in all patients with a previous history of goiter or nodules.

Environmental iodine intake affects the type of nonmalignant thyroid disease.

The relationship between the iodine intake level of a population and the occurrence of thyroid diseases is U-shaped with an increase in risk from both low and high iodine intakes. Developmental brain disorders and endemic goiter caused by severe iodine deficiency may seriously deteriorate overall health status and economic performance of a population. Severe iodine deficiency with a median 24-hour urinary iodine excretion of the population below 25 microg needs immediate attention and correction. Less severe iodine deficiency with median urinary iodine excretion below 120 microg per 24 hours is associated with multinodular autonomous growth and function of the thyroid gland leading to goiter and hyperthyroidism in middle aged and elderly subjects. The lower the iodine intake, the earlier and more prominent are the abnormalities. At the other end of the spectrum, severely excessive iodine intake starting at median urinary iodine excretion levels around 800 microg per 24 hours is associated with a higher prevalence of thyroid hypofunction and goiter in children. A number of studies indicate that moderate and mild iodine excess (median urinary iodine >220 microg per 24 hours) are associated with a more frequent occurrence of hypothyroidism, especially in elderly subjects. The exact mechanism leading to this has not been clarified, and more studies are needed to define the limits of excessive iodine intake precisely. Due to the frequent occurrence of thyroid disorders, proper monitoring and control of the population iodine intake level is a cost-effective alternative to diagnosing, therapy and control of the many individual cases of thyroid diseases that might have been prevented.

Validation of ultrasonography of the thyroid gland for epidemiological purposes.

Ultrasonography of the thyroid is often used in epidemiological surveys, thus thorough characterization of the interobserver variation of the different parameters obtained is important. Various methods have been used for measuring thyroid volume, and different formulas have been used for calculation of thyroid volume from the measured dimensions. In this article, two principles of thyroid volume measurement are described in detail: the wellknown method based on the three axes of each lobe and a new principle based on planimetry in two planes. The interobserver variation of the examination and the measuring procedure in itself were tested on 25 participants in a population study. A comparison of postmortem ultrasonography of the thyroid and results of an autopsy was performed. Good correlation and agreement between observers was found for thyroid volume (r = 0.98) and prevalence of thyroid nodules (kappa = 0.72), whereas echogenecity and echopattern showed little agreement. The correlation of thyroid volume by ultrasonography to autopsy results was satisfactory (r = 0.93), but the volume tended to be slightly underestimated even when using the formula pi/6(= 0.52)*length*width*depth. No major differences were found between the performance of the two principles of volume calculation. We conclude that when the measuring procedure is well defined, results of ultrasonography are comparable between observers for thyroid volume and prevalence of thyroid nodules, but not for echogenecity or echopattern. The formula of length*depth*width*pi/6 is suitable for thyroid volume measurement.

Thyroid disorders in mild iodine deficiency.

Comparative epidemiologic studies in areas with low and high iodine intake and controlled studies of iodine supplementation have demonstrated that the major consequence of mild-to-moderate iodine deficiency for the health of the population is an extraordinarily high occurrence of hyperthyroidism in elderly subjects, especially women, with risk of cardiac arrhythmias, osteoporosis, and muscle wasting. The hyperthyroidism is caused by autonomous nodular growth and function of the thyroid gland and it is accompanied by a high frequency of goiter. Pregnant women and small children are not immediately endangered but the consequences of severe iodine deficiency for brain development are grave and a considerable safety margin is advisable. Moreover, a shift toward less malignant types of thyroid cancer and a lower radiation dose to the thyroid in case of nuclear fallout support that mild-to-moderate iodine deficiency should be corrected. However, there is evidence that a high iodine intake may be associated with more autoimmune hypothyroidism, and that Graves' disease may manifest at a younger age and be more difficult to treat. Hence, the iodine intake should be brought to a level at which iodine deficiency disorders are avoided but not higher. Iodine supplementation programs should aim at relatively uniform iodine intake, avoiding deficient or excessive iodine intake in subpopulations. To adopt such a strategy, surveillance programs are needed.

Age- and sex-adjusted iodine/creatinine ratio. A new standard in epidemiological surveys? Evaluation of three different estimates of iodine excretion based on casual urine samples and comparison to 24 h values.

OBJECTIVE: The most accurate way to measure urinary iodine excretion in epidemiological surveys is still debated. We propose a new principle of estimating iodine excretion based on casual urine samples. MATERIAL AND METHODS: A total of 123 24 h urine samples and corresponding casual urine samples were collected from 31 subjects. Iodine excretion was expressed as 24 h iodine excretion and three different estimates: iodine concentration in the casual sample, iodine/gram creatinine in the casual sample, and the new principle-iodine/creatinine ratio in the casual sample, adjusted for expected creatinine excretion of the individual. RESULTS: All three estimates based on casual urine samples correlated significantly to 24 h values with a r (Pearson) of 0.37 for iodine concentration, 0. 61 for iodine/creatinine ratio and 0.62 for the age- and sex-adjusted iodine/creatinine ratio. The median iodine excretion in the entire group was 143 microg/day in 24 h samples, 87 microg/l as iodine concentration, 77 microg/g creatinine as iodine/creatinine ratio and 126 microg/day as age- and sex-adjusted iodine/creatinine ratio. CONCLUSION: Age- and sex-adjusted iodine/creatinine ratio is a more accurate and unbiased estimate of iodine excretion in epidemiological surveys of adults than the two most frequently used estimated: iodine concentration and iodine/gram creatinine, as these two estimates may introduce a bias depending on the composition of the investigated group. The adjusted iodine/creatinine ratio is superior to the other estimates, especially when individual estimates of 24 h iodine excretion is required or cohorts of selected groups are investigated. SPONSORSHIP: This work was supported by grants from the Medical Research Foundation Region Greater Copenhagen, Faroe Islands and Greenland; the Wedell-Wedellsborg Foundation; Musikforlaeggerne Agnes and Knut Morks Foundation.

Evaluation of a semi-quantitative food frequency questionnaire to estimate iodine intake.

OBJECTIVE: To evaluate a food frequency questionnaire (FFQ) used to assess the dietary intake of iodine. DESIGN: The iodine intake determined by the FFQ was compared with 4-day dietary records and with iodine excretion in 24 h urine samples in a subgroup of participants in a cross-sectional study of iodine intake and thyroid diseases in Denmark. Furthermore, the intake of fish determined from the FFQ was compared with the intake of fish from a simple record kept for 3 months. SUBJECTS: Women aged 25-30 y and 60-65 y. RESULTS: Median iodine intake was similar when determined from the FFQ and from dietary records and the correlation between these measures was 0.52 (P < 0.001). Iodine intake was higher than iodine excretion (P < 0.001). The cross-check questions in the FFQ (for example the question 'How often did you get any kind of fish?') underestimated the intake. In contrast, the intake of a specific fish tended to be overestimated by the FFQ when compared with the 3 month record of fish intake. CONCLUSION: The FFQ can be used to classify subjects into low and high iodine intake groups, but the level of iodine tends to be overestimated.

Heart rate variability and n-3 fatty acids in patients with chronic renal failure--a pilot study.

Patients with chronic renal failure (CRF) often have autonomic cardiac dysfunction, which can be assessed by measuring heart rate variability (HRV). This dysfunction prediposes the patients to sudden cardiac death. This study describes 24-hour HRV in patients with CRF compared to HRV in patients with a previous myocardial infarction (MI). Furthermore, associations between HRV in patients with CRF and the content of n-3 polyunsaturated fatty acids (PUFA) in cell membranes were examined, because n-3 PUFA may improve HRV. Twenty-nine patients with CRF treated with dialysis were enrolled. A 24-hour Holter recording was obtained at baseline and the HRV variables, RR (= mean of all normal RR intervals during the 24-hour recording) and SDNN (= standard deviation of all normal RR intervals in the entire 24-hour recording) were analyzed. Also, granulocyte fatty acid composition was determined. The patients were allocated to dietary supplementation with either 5.2 g of n-3 PUFA or a placebo oil (olive oil) daily for 12 weeks in a double-blind design. At the end of the supplementation period the Holter recording and blood sampling were repeated. At baseline the CRF patients' mean SDNN ws 86 ms compared to 118 ms (p < 0.01) in patients with a previous MI. After supplementation with either n-3 PUFA or placebo a highly significant correlation was observed between the content of n-3 PUFA in cell membranes and HRV (r = 0.71, p < 0.01). Furthermore, when the patients were dichotomized according to their mean SDNN, it was found, that those with the highest SDNN had a higher content of n-3 PUFA in cell membranes compared to those with the lowest SDNN (7.8% vs 4.2%, p < 0.05). In conclusion, HRV was decreased in CRF patients indicating a cardiovascular autonomic dysfunction. The positive correlation between the n-3 PUFA content in cell membranes and HRV suggests that the effects of an increased intake of n-3 PUFA in CRF patients should be further studied.

Malignant pleural mesothelioma.

Malignant pleural mesotheliomas are tumors originating in the serosal lining of the pleural cavities. Although these are relatively rare tumors, an increasing number of cases are anticipated over the next decade. Patients with these tumors, which are almost invariably fatal, usually have an interval of less than two years between the onset of symptoms and death. There is a well established link between mesothelioma and asbestos exposure. Treatment includes surgery, chemotherapy, and radiotherapy. This article explores the epidemiology and etiology of the disease, offers a summary of current treatment options, and discusses nursing strategies.

Liposarcoma of the thyroid gland.

The presumed first case of primary liposarcoma of the thyroid gland is presented, and the possible origin of this rare tumor from either mature fat cells or primitive mesenchymal cells is discussed.

[Licorice. A basis for precautions one more time!]. / Lakrids. Grund til at advare endnu en gang!

A 38 year-old male was admitted to hospital with somnolence, flaccid paralysis of the extremities, arterial hypertension, oedema, severe hypokalemia and rabdomyolysis. The course was complicated with respiratory and kidney failure. It became apparent that the symptoms were caused by the ingestion of 200 g of licorice daily for ten weeks together with a thiazide diuretic for two weeks.