Redox balance and immunity of piglets pre- and post-E. coli challenge after treatment with hemp or fish oil, and vitamin E.

This study investigated the influence of polyunsaturated fatty acid composition and vitamin E supplementation on oxidative status and immune responses in weanling piglets pre- and post-E. coli challenge. Suckling piglets (n = 24) were randomly selected from two litters for an oral supplementation (1 mL/day) with fish oil or hemp oil and vitamin E supplementation (60 mg natural vitamin E/mL oil) from day 10 to 28 of age. At day 29 and 30 of age, each piglet was orally inoculated with 6.7 × 108 and 3.96 × 108 CFU of F4 and F18 E. coli, respectively. Blood was sampled from all piglets on day 28 before E. coli challenge and on day 35 of age to investigate immunological and oxidative stress markers in plasma. One week after weaning and exposure to E. coli, a general reduction in the α-tocopherol concentration and activity of GPX1 was obtained. Vitamin E supplementation lowered the extent of lipid peroxidation and improved the antioxidative status and immune responses after E. coli challenge. Hemp oil had the greatest effect on antioxidant enzyme activity. Provision of hemp oil and vitamin E to suckling piglets may reduce the incidence of post-weaning diarrhea.

Frequency of misdiagnosis in hypertrophic cardiomyopathy.

BACKGROUND: Hypertrophic Cardiomyopathy (HCM) is characterized by unexplained left ventricle hypertrophy (LVH) ≥15 mm. The condition is often hereditary and family screening is recommended to reduce the risk of adverse disease complications and premature death among relatives. Correct diagnosis of index patients is important to ensure that only relatives at risk of disease development are invited for family screening. PURPOSE: To investigate if patients with ICD-10 codes for HCM (DI421) or hypertrophic obstructive cardiomyopathy (DI422) fulfilled recognised diagnostic criteria. METHODS: All patients with ICD-10 codes for HCM or HOCM at a Department of Cardiology were identified and had their diagnosis validated by a cardiac investigation or a review of their medical records and previous investigations. RESULTS: Two hundred and forty patients had ICD-10 codes for HCM/HOCM, of whom 202 (84%, 202/240) underwent re-examination, while 38 (16%, 38/240) had their hospital notes reviewed. Seventy-six patients (32%, n = 76/240) did not fulfil diagnostic criteria, of whom 39, (51%, n = 39/76) had normal (10 mm) or modest LV wall thickness (11-14 mm). The remaining 37 patients (49%, n = 37/76) had LVH ≥15 mm, which was well-explained by uncontrolled hypertension, (32%, n = 24/76), aortic valve stenosis (19%, n = 7/76) or wild-type amyloidosis (16%, 6/76). CONCLUSION: One-third of patients with ICD-10 codes for HCM or HOCM did not fulfil recognised diagnostic criteria. Incorrect diagnosis of HCM may cause unnecessary family investigations which may be associated with anxiety, and a waste of health care resources. This highlights the need for specialised cardiomyopathy services to ensure correct diagnosis and management of HCM.

The time course of feature-selective attention inside and outside the focus of spatial attention.

Research on attentional selection of stimulus features has yielded seemingly contradictory results. On the one hand, many experiments in humans and animals have observed a "global" facilitation of attended features across the entire visual field, even when spatial attention is focused on a single location. On the other hand, several event-related potential studies in humans reported that attended features are enhanced at the attended location only. The present experiment demonstrates that these conflicting results can be explained by differences in the timing of attentional allocation inside and outside the spatial focus of attention. Participants attended to fields of either red or blue randomly moving dots on either the left or right side of fixation with the task of detecting brief coherent motion targets. Recordings of steady-state visual evoked potentials elicited by the flickering stimuli allowed concurrent measurement of the time course of feature-selective attention in visual cortex on both the attended and the unattended sides. The onset of feature-selective attentional modulation on the attended side occurred around 150 ms earlier than on the unattended side. This finding that feature-selective attention is not spatially global from the outset but extends to unattended locations after a temporal delay resolves previous contradictions between studies finding global versus hierarchical selection of features and provides insight into the fundamental relationship between feature-based and location-based (spatial) attention mechanisms.

Proinflammatory polarization strongly reduces human macrophage in vitro phagocytosis of tumor cells in response to CD47 blockade.

Antibody-based CD47 blockade aims to activate macrophage phagocytosis of tumor cells. However, macrophages possess a high degree of phenotype heterogeneity that likely influences phagocytic capacity. In murine models, proinflammatory (M1) activation increases macrophage phagocytosis of tumor cells, but in human models, results have been conflicting. Here, we investigated the effects of proinflammatory polarization on the phagocytic response of human monocyte-derived macrophages in an in vitro model. Using both flow cytometry-based and fluorescence live-cell imaging-based phagocytosis assays, we observed that mouse monoclonal anti-CD47 antibody (B6H12) induced monocyte-derived macrophage phagocytosis of cancer cells in vitro. Proinflammatory (M1) macrophage polarization with IFN-γ+LPS resulted in a severe reduction in phagocytic response to CD47 blockade. This reduction coincided with increased expression of the antiphagocytic membrane proteins LILRB1 and Siglec-10 but was not rescued by combination blockade of the corresponding ligands. However, matrix metalloproteinase inhibitors (TAPI-0 or GM6001) partly restored response to CD47 blockade in a dose-dependent manner. In summary, these data suggest that proinflammatory (M1) activation reduces phagocytic response to CD47 blockade in human monocyte-derived macrophages.

A high fat to vitamin E ratio in the feed protects and improves uptake of the natural form of vitamin E in postweaning calves.

In postweaning calves, it is a challenge to maintain the plasma vitamin E level at or above the recommended level (3 µg/mL), which is linked to a good immune response. It has been unclear until now why the provision of solid feed with concentrations below 200 mg/kg feed of vitamin E is ineffective in maintaining the plasma vitamin E level of calves above the recommended plasma level postweaning. The present study was conducted to investigate if a high fat to vitamin E ratio in the concentrate could protect and improve the delivery of the natural form of vitamin E (RRR-α-tocopherol) to calves postweaning. Thirty calves were included in the experiment from 2 weeks preweaning until 2 weeks postweaning (Weeks -2, -1, 0 [weaning], 1, and 2 relative to weaning) and fed one of three concentrates in which lecithin mixture provided the fat supplement: control (77 mg/kg of vitamin E and 4.9% DM of crude fat; CONT), medium level of vitamin E supplemented (147 mg/kg of vitamin E and 7.7% DM of crude fat; MedVE) or high level of vitamin E supplemented (238 mg/kg of vitamin E and 12.4% DM of fat; HiVE). Thus, there was a comparable ratio of fat to vitamin E (520-630) in the three concentrates. During the 2 weeks postweaning, final body weight (92 ± 2 kg), average daily gain (917 ± 51 g/day) and concentrate intake (2.2 ± 0.09 kg/day; mean of treatment ± standard error) were unaffected by treatment and the interaction between treatment and week. There was an interaction between treatment and week for vitamin E intake pre- (p < 0.001) and postweaning (p < 0.001). There was an interaction between treatment and week (p < 0.001) for plasma vitamin E level postweaning, and it was 2.5, 3.1, and 3.8 µg/mL in CONT, MedVE, and HiVE, respectively, at Week 1 postweaning. In addition, plasma vitamin E levels at Week 2 postweaning were 2.6, 3.6 and 4.8 µg/mL in CONT, MidVE and HiVE respectively. The results show that 147 mg/kg of lecithin-protected vitamin E in the concentrate is needed to secure a plasma vitamin E level well above the recommended level. In addition, lecithin-protected vitamin E elevated the plasma level of triglycerides and nonesterified fatty acids.

Clinical Frailty Scale at presentation to the emergency department: interrater reliability and use of algorithm-assisted assessment.

PURPOSE: The Clinical Frailty Scale (CFS) allows health care providers to quickly stratify older patients, to support clinical decision-making. However, few studies have evaluated the CFS interrater reliability (IRR) in Emergency Departments (EDs), and the freely available smartphone application for CFS assessment was never tested for reliability. This study aimed to evaluate the interrater reliability of the Clinical Frailty Scale (CFS) ratings between experienced and unexperienced staff (ED clinicians and a study team (ST) of medical students supported by a smartphone application to assess the CFS), and to determine the feasibility of CFS assignment in patients aged 65 or older at triage. METHODS: Cross-sectional study using consecutive sampling of ED patients aged 65 or older. We compared assessments by ED clinicians (Triage Clinicians (TC) and geriatric ED trained nurses (geriED-TN)) and a study team (ST) of medical students using a smartphone application for CFS scoring. The study is registered on Clinicaltrials.gov (NCT05400707). RESULTS: We included 1349 patients aged 65 and older. Quadratic-weighted kappa values for ordinal CFS levels showed a good IRR between TC and ST (Ï° = 0.73, 95% CI 0.69-0.76), similarly to that between TC and geriED-TN (Ï° = 0.75, 95% CI 0.66-0.82) and between the ST and geriED-TN (Ï° = 0.74, 95% CI 0.63-0.81). A CFS rating was assigned to 972 (70.2%) patients at triage. CONCLUSION: We found good IRR in the assessment of frailty with the CFS in different ED providers and a team using a smartphone application to support rating. A CFS assessment occurred in more than two-thirds (70.2%) of patients at triage.

Attentional Modulation in Early Visual Cortex: A Focused Reanalysis of Steady-state Visual Evoked Potential Studies.

Steady-state visual evoked potentials (SSVEPs) are a powerful tool for investigating selective attention. Here, we conducted a combined reanalysis of multiple studies employing this technique in a variety of attentional experiments to, first, establish benchmark effect sizes of attention on amplitude and phase of SSVEPs and, second, harness the power of a large data set to test more specific hypotheses. Data of eight published SSVEP studies were combined, in which human participants (n = 135 in total) attended to flickering random dot stimuli based on their defining features (e.g., location, color, luminance, or orientation) or feature conjunctions. The reanalysis established that, in all the studies, attention reliably enhanced amplitudes, with color-based attention providing the strongest effect. In addition, the latency of SSVEPs elicited by attended stimuli was reduced by ∼4 msec. Next, we investigated the modulation of SSVEP amplitudes in a subset of studies where two different features were attended concurrently. Although most models assume that attentional effects of multiple features are combined additively, our results suggest that neuronal enhancement provided by concurrent attention is better described by multiplicative integration. Finally, we used the combined data set to demonstrate that the increase in trial-averaged SSVEP amplitudes with attention cannot be explained by increased synchronization of single-trial phases. Contrary to the prediction of the phase-locking account, the variance across trials of complex Fourier coefficients increases with attention, which is more consistent with boosting of a largely phase-locked signal embedded in non-phase-locked noise.

Sex-dimorphic neuroprotective effect of CD163 in an α-synuclein mouse model of Parkinson's disease.

Alpha-synuclein (α-syn) aggregation and immune activation represent hallmark pathological events in Parkinson's disease (PD). The PD-associated immune response encompasses both brain and peripheral immune cells, although little is known about the immune proteins relevant for such a response. We propose that the upregulation of CD163 observed in blood monocytes and in the responsive microglia in PD patients is a protective mechanism in the disease. To investigate this, we used the PD model based on intrastriatal injections of murine α-syn pre-formed fibrils in CD163 knockout (KO) mice and wild-type littermates. CD163KO females revealed an impaired and differential early immune response to α-syn pathology as revealed by immunohistochemical and transcriptomic analysis. After 6 months, CD163KO females showed an exacerbated immune response and α-syn pathology, which ultimately led to dopaminergic neurodegeneration of greater magnitude. These findings support a sex-dimorphic neuroprotective role for CD163 during α-syn-induced neurodegeneration.

Formation of RRR-α-tocopherol in rumen and intestinal digestibility of tocopherols in dairy cows.

Tocopherol sources in diets are often a combination of all-rac-α-tocopheryl acetate (synthetic α-tocopherol) from vitamin supplements and natural tocopherols and 2R-(4'R, 8'R)-5,7,8-trimethyltocotrienol (α-tocotrienols) from the feed sources. Synthetic α-tocopherol consists of 8 different stereoisomers including 2R-(4'R, 8'R)-5,7,8-trimethyltocol (RRR-α-tocopherol), 2R-(4'S, 8'R)-5,7,8-trimethyltocol (RSR-α-tocopherol), 2R-(4'R, 8'S)-5,7,8-trimethyltocol (RRS-α-tocopherol), 2R-(4'S, 8'S)-5,7,8-trimethyltocol (RSS-α-tocopherol), 2S-(4'S, 8'S)-5,7,8-trimethyltocol (SSS-α-tocopherol), 2S-(4'R, 8'S)-5,7,8-trimethyltocol (SRS-α-tocopherol), 2S-(4'S, 8'R)-5,7,8-trimethyltocol (SSR-α-tocopherol), and 2S-(4'R, 8'R)-5,7,8-trimethyltocol (SRR-α-tocopherol). The pre-absorption metabolism of tocopherols and tocotrienols in ruminants differs from monogastric animals due to the extensive microbial fermentation in the anaerobic rumen. The current study investigated the impact of toasting and decortication of oats on metabolism in the digestive tract (synthesis, digestion), and intestinal digestibility of tocopherols in dairy cows by using 4 ruminal and intestinal cannulated Danish Holstein cows in a 4 × 4 Latin square design for 4 periods. Cows were fed a total mixed ration ad libitum containing different forms of oats: whole oat, decorticated oat, toasted oat, and decorticated toasted oat, all rolled before mixed ration. Overall means across 4 treatments were statistically analyzed, testing whether overall means were different from zero. Decortication or toasting did not affect the balance or digestibility of α-tocopherols in rumen. Average across treatments showed the ruminal degradation of synthetic α-tocopherol (279 mg/d, P = 0.02; P-value shows that average across treatments is different from zero), synthetic 2R-α-tocopherol (133 mg/d, P < 0.01; summation of RRS-, RSR- and RSS-α-tocopherol), and 2S-α-tocopherol (190 mg/d; P < 0.01, summation of SSS-, SRS-, SSR, and SRR-α-tocopherol), while RRR-α-tocopherol was formed in the rumen (221 mg/d, P = 0.10). The average across treatments showed that small intestinal digestibility of tocopherols ranked in the following order: α-tocotrienol > natural α-tocopherol > synthetic α-tocopherols > 2R-(4'R, 8'R)-,7,8-dimethyltocol (γ-tocopherol). The average across treatments for small intestinal and feed-ileum digestibility ranked in the following order: RRR-α-tocopherol > synthetic 2R-α-tocopherol > 2S-α-tocopherol. Results showed the first evidence for RRR-α-tocopherol formation under anaerobic conditions in the rumen. In addition, synthetic α-tocopherol stereoisomers, γ-tocopherol and α-tocotrienol were degraded in the rumen. There was a discrimination against absorption of synthetic 2R- and 2S-α-tocopherol in the small intestine.

The effect of dietary omega-3 fatty acid supplementation on fetal growth, piglet birth weight and plasma fatty acid concentrations, using docosahexaenoic acid in early gestation in sows.

The objective of the study was to test the effect of the omega-3 fatty acid docosahexaenoic acid (DHA) on fetal and placental development as well as the birth weight of piglets. A total of 238 multiparous sows were allocated to either a control diet group or a DHA diet group with an omega-6 to omega-3 ratio of 9.8 and 2.4, respectively, from mating to day 43 of gestation. A blood sample was collected and back fat thickness was measured prior to mating, on days 14, 42 and 112 of gestation. On day 43 of gestation, 14 sows were slaughtered and measurements of fetuses and placentas were taken. Piglets in some litters were weighed individually at farrowing. Dietary treatment did not affect fetal characteristics and back fat thickness (P > 0.05). Dietary treatment increased the plasma concentrations of total omega-3 fatty acids in sows (P < 0.05). Sows fed the DHA diet had a shorter gestation length compared to the control sows (P < 0.05), but the number of born piglets was not affected (P > 0.05). The average piglet birth weight and the within-litter variation in birthweight were unaffected by dietary DHA (P > 0.05), however, sows fed DHA diet had fewer piglets under 800 g at birth compared to control sows (P < 0.05). In conclusion, addition of DHA decreased the dietary ratio of omega-6 to omega-3 fatty acids, increased plasma n-3 fatty acid concentrations in sows and decreased the number of piglets weighing under 800 g at birth.

Supplementation of palmitoleic acid improved piglet growth and reduced body temperature drop upon cold exposure.

Piglet survival is a major challenge in the first few days postpartum and interventions during this period may improve survival and growth. This study investigated the effects of palmitoleic acid (C16:1n-7; PA) supplementation on growth performance, body temperature, fatty acid (FA), and energy metabolism in milk-replacer-fed piglets. Forty-eight piglets were stratified by body weight and randomly assigned to one of four dietary treatments (0%, 1%, 2%, and 3% PA supplementation as a percent of milk replacer) and given the diet through an orogastric tube. They were fed dietary treatments every 2 h for 4 d in the first week postpartum and all were sacrificed at the end of the experiment. The piglets were weighed daily, and half in each dietary treatment group, the same piglets each day, were exposed daily to a lower temperature for 2 h. Plasma samples were collected immediately before sacrifice for analyses of FA and other plasma metabolites. The weight of organs and empty body weight were determined after sacrifice. Liver and semimembranosus muscle tissue samples were collected and analyzed for FA content. Contents of C16:1n-7 and C18:1n-7 in both plasma and liver (P < 0.001), and C16:1n-7 in semimembranosus muscle (P < 0.001) increased linearly as PA supplementation increased. Most plasma FA levels (except C16:1n-7, C16:1n-9, and C22:5n-3) were lower in piglets exposed to lower temperatures than those that were not. Plasma glucose, triglycerides, and lactate dehydrogenase levels increased linearly with PA supplementation (P < 0.001). Piglets' average daily gain, liver glycogen pool, liver weight, and gallbladder weight increased linearly (P < 0.05, P < 0.01, P < 0.05, and P < 0.001, respectively), but lung weight, liver nitrogen content, and body temperature drop decreased linearly (P < 0.01, P < 0.001, and P < 0.05, respectively) with PA supplementation. Piglets exposed to low temperature had greater liver nitrogen (P < 0.05) and lactate dehydrogenase (P < 0.001) contents but had lower liver weight (P < 0.01) and plasma lactate concentration (P < 0.05) than those that were not. In conclusion, this study demonstrated the importance of PA on the growth performance of the piglets by increasing their average daily gain and decreasing a drop in body temperature upon cold exposure, most likely due to a modified energy metabolism.

Reducing piglet mortality in the early days after birth is a significant challenge in the modern pig industry. The focus on achieving larger litter sizes has had a negative impact on piglets' birth weight and their intake of colostrum. Additionally, piglets are born without easily oxidizable brown adipose tissue and have limited body reserves, making them more vulnerable to death due to their lower capacity for thermogenesis. Therefore, it is important to explore dietary strategies that can enhance piglets' thermogenesis capacity. In this study, the role of palmitoleic acid supplementation was investigated in a dose-response design to determine its impact on growth performance, fatty acid composition, and energy metabolism of milk-replacer-fed piglets during their first week of life. The results revealed a linear increase in the average daily gain of the piglets, liver weight, and liver glycogen content with increasing palmitoleic acid supplementation. Moreover, increased palmitoleic acid supplementation was associated with a drop in body temperature when piglets were exposed to a lower temperature during the experimental period. Altogether, the study indicated that palmitoleic acid has a sparing effect on glycogen reserves and that a greater proportion of energy utilized by the piglets to maintain their body temperature was derived from the oxidation of fatty acids. The results indicated a promising approach to improve piglet survival and growth through dietary modifications of fatty acids in the diet.

Genetic mapping of quantitative trait loci associated with drought tolerance in chickpea (Cicer arietinum L.).

Elucidation of the genetic basis of drought tolerance is vital for genomics-assisted breeding of drought tolerant crop varieties. Here, we used genotyping-by-sequencing (GBS) to identify single nucleotide polymorphisms (SNPs) in recombinant inbred lines (RILs) derived from a cross between a drought tolerant chickpea variety, Pusa 362 and a drought sensitive variety, SBD 377. The GBS identified a total of 35,502 SNPs and subsequent filtering of these resulted in 3237 high-quality SNPs included in the eight linkage groups. Fifty-one percent of these SNPs were located in the genic regions distributed throughout the genome. The high density linkage map has total map length of 1069 cm with an average marker interval of 0.33 cm. The linkage map was used to identify 9 robust and consistent QTLs for four drought related traits viz. membrane stability index, relative water content, seed weight and yield under drought, with percent variance explained within the range of 6.29%-90.68% and LOD scores of 2.64 to 6.38, which were located on five of the eight linkage groups. A genomic region on LG 7 harbors quantitative trait loci (QTLs) explaining > 90% phenotypic variance for membrane stability index, and > 10% PVE for yield. This study also provides the first report of major QTLs for physiological traits such as membrane stability index and relative water content for drought stress in chickpea. A total of 369 putative candidate genes were identified in the 6.6 Mb genomic region spanning these QTLs. In-silico expression profiling based on the available transcriptome data revealed that 326 of these genes were differentially expressed under drought stress. KEGG analysis resulted in reduction of candidate genes from 369 to 99, revealing enrichment in various signaling pathways. Haplotype analysis confirmed 5 QTLs among the initially identified 9 QTLs. Two QTLs, qRWC1.1 and qYLD7.1, were chosen based on high SNP density. Candidate gene-based analysis revealed distinct haplotypes in qYLD7.1 associated with significant phenotypic differences, potentially linked to pathways for secondary metabolite biosynthesis. These identified candidate genes bolster defenses through flavonoids and phenylalanine-derived compounds, aiding UV protection, pathogen resistance, and plant structure.The study provides novel genomic regions and candidate genes which can be utilized in genomics-assisted breeding of superior drought tolerant chickpea cultivars.

Patients With Hypertrophic Cardiomyopathy and Normal Genetic Investigations Have Few Affected Relatives.

BACKGROUND: Current guidelines recommend that relatives of index patients with hypertrophic cardiomyopathy (HCM) are offered clinical investigations to identify individuals at risk of adverse disease complications and sudden cardiac death. However, the value of family screening in relatives of index patients with a normal genetic investigation of recognized HCM genes is largely unknown. OBJECTIVES: The purpose of this study was to perform family screening among relatives of HCM index patients with a normal genetic investigation to establish the frequency of familial disease and the clinical characteristics of affected individuals. METHODS: Clinical and genetic investigations were performed in consecutive and unrelated HCM index patients. Relatives of index patients who did not carry pathogenic/likely pathogenic variants in recognized HCM genes were invited for clinical investigations. RESULTS: In total, 60% (270 of 453) of HCM index patients had a normal genetic investigation. A total of 80% of their relatives (751 of 938, median age 44 years) participated in the study. Of these, 5% (34 of 751) were diagnosed with HCM at baseline, whereas 0.3% (2 of 717 [751-34]) developed the condition during 5 years of follow-up. Their median age at diagnosis was 57 years (IQR: 51-70 years). Two-thirds (22 of 36) were diagnosed following family screening, whereas one-third (14 of 36) had been diagnosed previously because of cardiac symptoms, a murmur, or an abnormal electrocardiogram. None of the affected relatives experienced adverse disease complications. The risk of SCD was low. CONCLUSIONS: Systematic family screening of index patients with HCM and normal genetic investigations was associated with a low frequency of affected relatives who appeared to have a favorable prognosis.

Vegetal protein hydrolysates reduce the yield losses in off-season crops under combined heat and drought stress.

To deal with the vagaries of climate change, it is essential to develop climate-resilient agricultural practices, which improve crop productivity, and ensure food security. The impacts of high temperature and water deficit stress conditions pose serious challenges to a sustainable crop production. Several adaptation measures are practiced globally to address these challenges and among these altering the crop's typical growing season is one of the key management practices. Application of biostimulants and other growth hormones helps in compensating yield losses under abiotic stress significantly. Therefore, this study was conducted to evaluate the influence of vegetal protein hydrolysate based biostimulant to reduce the yield losses of off-season crops (soybean and chilli in summer and chickpea in early Kharif) when the temperature was higher than the regular season under water deficit stress conditions. The experiments were carried out with the foliar application of different protein hydrolysates (PHs) concentrations. The study revealed that the application of PHs significantly improved the membrane stability index, relative water content, total chlorophyll and proline content of leaves. Consequently, it led to an increase in the number of pods in soybean and chickpea, and fruits in chilli, leading to improved yields when plants were treated with the appropriate amount of PHs. Compared to untreated plants, PHs helped improve the efficiency of PS-II with significantly high photochemical efficiency (QYmax) even at higher excised leaf water loss or reduction in loss of relative water content. This study concluded that foliar application of PHs at 4, 2, and 6 ml L-1 can be beneficial for soybean, chickpea and chilli, which exhibited 17, 30, and 25% yield improvement respectively, over the untreated plants under water deficit stress. It is suggested that the benefits of PHs can be realized in soybean, chickpea and chilli under high temperature and water deficit stress. Therefore, vegetal PHs may be able to assist farmers in arid regions for boosting their income by raising market value and decreasing production barriers during the off-season. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01334-4.

Stage-Specific Risk of Recurrence and Death From Melanoma in Denmark, 2008-2021: A National Observational Cohort Study of 25 720 Patients With Stage IA to IV Melanoma.

Importance: To ensure optimal treatment and surveillance of patients with melanoma, knowledge of the clinical stage-specific risk of recurrence, mortality, and recurrence patterns across the American Joint Committee on Cancer Eighth Edition (AJCC8) substages is needed. Objective: To estimate stage-specific recurrence and melanoma-specific mortality rates, assess absolute stage-specific risks of recurrence and mortality, and describe stage-specific recurrence patterns, including conditional rates. Design: Retrospective cohort study of prospectively collected nationwide population-based registry data. Setting: Nationwide, population-based cohort study. Participants: The 25 720 Danish patients, 18 years or older, diagnosed with first-time stage IA to IV cutaneous melanoma between January 1, 2008, and December 31, 2019, were included and followed up from time of primary treatment until December 31, 2021. Exposures: First diagnosis of stage IA to IV cutaneous melanoma. Main Outcomes: Stage-specific cumulative incidence of recurrence and melanoma-specific mortality, melanoma-specific recurrence-free survival, and assessed absolute stage-specific risks of recurrence and melanoma-specific mortality. Secondary outcomes were stage-specific recurrence patterns, including conditional rates, and melanoma-specific survival. Results: We followed up 25 720 patients for a median of 5.9 years (95% CI, 58.9-59.3 years). Mean age was 59.1 years (95% CI, 58.9-59.3 years). Patients with stage IIB to IIC melanoma were older, had more comorbidities at diagnosis, and had the lowest rate of pathologic staging by sentinel node biopsy (81.6%-87.4%). A total of 10.6% of patients developed recurrence; first recurrence included distant recurrence, alone or with synchronous locoregional recurrence, in 56.6% of patients. We found a comparable risk of recurrence in stages IIIA and IIB (29.7% vs 33.2%) and in stages IIIB and IIC (35.9% vs 36.8%), respectively. Melanoma-specific mortality was comparable between stages IIIA and IIA (13.0% vs 13.6%) and between stages IIIB and IIB (18.4% vs 22.0%), respectively. These risk patterns persisted in cause-specific hazards models. Conclusions and Relevance: This nationwide, population-based cohort study found that the increasing stages of the current AJCC8 staging system do not accurately reflect an increasing risk of recurrence and mortality in melanoma. The high proportion of distant recurrences suggests that hematogenous spread is a more common metastatic pathway than previously assumed, and surveillance with routine functional/cross-sectional imaging should be considered for stages IIB to IV. Future efforts should be put toward developing new tools for risk stratification and determining the survival effect of routine imaging in surveillance.

Comparison of culture media reveals that non-essential amino acids strongly affect the phenotype of human monocyte-derived macrophages.

Macrophages are important innate immune cells with the ability to adapt their phenotype to environmental cues. Research on human macrophages often uses monocyte-derived macrophages cultured in vitro, but it is unclear if culture medium affects macrophage phenotype. The objective of this study was to determine the impact of culture medium composition on monocyte-derived macrophage phenotype. Monocyte-derived macrophages were generated in different formulations of culture media (RPMI 1640, DMEM, MEM, McCoy's 5a and IMDM). Viability, yield and cell size were monitored, and RT-qPCR, flow cytometry or ELISA was used to compare levels of phenotype markers (CD163, CD206, CD80, TNFα, IL-10, SIRPα, LILRB1 and Siglec-10). Yield, cell size, gene expression, membrane protein levels and release of soluble proteins were all affected by changes in culture medium composition. The most pronounced effects were observed after culture in DMEM, which lacks the non-essential amino acids asparagine, aspartic acid, glutamic acid and proline. Supplementation of DMEM with non-essential amino acids either fully or partly reversed most effects of DMEM on macrophage phenotype. The results suggest culture medium composition and amino acid availability affect the phenotype of human monocyte-derived macrophages cultured in vitro.

Structured decision support to prevent hospitalisations of community-dwelling older adults in Denmark (PATINA): an open-label, stepped-wedge, cluster-randomised controlled trial.

BACKGROUND: Ageing populations and health-care staff shortages encourage efforts in primary care to recognise and prevent health deterioration and acute hospitalisation in community-dwelling older adults. The PATINA algorithm and decision-support tool alerts home-based-care nurses to older adults at risk of hospitalisation. The study aim was to test whether use of the PATINA tool was associated with changes in health-care use. METHODS: An open-label, stepped-wedge, cluster-randomised controlled trial was done in three Danish municipalities, covering 20 area teams providing home-based care to around 7000 recipients. During a period of 12 months, area teams were randomly assigned to an intervention crossover for older adults (aged 65 years or older) who received care at home. The primary outcome was hospitalisation within 30 days of identification by the algorithm as being at risk of hospitalisation. Secondary outcomes were hospital readmission and other hospital contacts, outpatient contacts, contact with primary care physicians (PCPs), temporary care, and death, within 30 days of identification. This study was registered at ClinicalTrials.gov (NTC04398797). FINDINGS: In total, 2464 older adults participated in the study: 1216 (49·4%) in the control phase and 1248 (50·6%) in the intervention phase. In the control phase, 102 individuals were hospitalised within 30 days during 33 943 days of risk (incidence 0·09 per 30 days), compared with 118 individuals within 34 843 days of risk (0·10 per 30 days) during the intervention phase. The intervention was not associated with a reduction in the number of first hospitalisations within 30 days (incidence rate ratio [IRR] 1·10 [90% CI 0·90-1·40]; p=0·28). Furthermore it was not associated with reduced rates of other hospital contacts (IRR 1·10 [95% CI 0·90-1·40]; p=0·28), outpatient contacts (1·10 [0·88-1·40]; p=0·42), or mortality (0·82 [0·58-1·20]; p=0·25). The intervention was associated with a 59% reduction in readmissions within 30 days of hospital discharge (IRR 0·41 [95% CI 0·24-0·68]; p=0·0007), a 140% increase in contacts with PCPs (2·40 [1·18-3·20]; p<0·0001), and a 150% increase in use of temporary care (2·50 [1·40-4·70]; p=0·0027). INTERPRETATION: Despite having no effect on the primary outcome, the PATINA tool showed other benefits for older adults receiving home-based care. Such algorithms have the potential to shift health-care use from secondary to primary care but need to be tested in other home-based care settings. Implementation of algorithms in clinical practice should be informed by analysis of cost-effectiveness and potential harms as well as the benefits. FUNDING: Innovation Fund Denmark and Region of Southern Denmark. TRANSLATIONS: For the Danish, French and German translations of the abstract see Supplementary Materials section.

The effect of air quality on sleep and cognitive performance in school children aged 10-12 years: a double-blinded, placebo-controlled, crossover trial.

OBJECTIVES: To investigate the effect of CO2 during sleep on next-morning cognitive performance in young schoolchildren, the authors performed a double-blind fully balanced crossover placebo-controlled study. MATERIAL AND METHODS: The authors included 36 children aged 10-12 years in the climate chamber. The children slept at 21°C in 6 groups each at 3 different conditions separated by 7 days in a random order. Conditions were as follows: high ventilation with CO2 at 700 ppm, high ventilation with added pure CO2 at 2000-3000 ppm, and reduced ventilation with CO2 at 2-3000 ppm and bioeffluents. Children were subjected to a digital cognitive test battery (CANTAB) in the evening prior to sleep and on the next morning after breakfast. Sleep quality was monitored with wrist actigraphs. RESULTS: There were no significant exposure effects on cognitive performance. Sleep efficiency was significantly lower at high ventilation with CO2 at 700 ppm which is considered to be a chance effect. No other effects were seen, and no relation between air quality during sleep and next-morning cognitive performance was observed in the children emitting an estimated 10 lCO2/h per child. CONCLUSIONS: No effect of CO2 during sleep was found on next day cognition. The children were awakened in the morning, and spent from 45-70 min in well-ventilated rooms before they were tested. Hence, it cannot be precluded that the children have benefitted from the good indoor air quality conditions before and during the testing period. The slightly better sleep efficiency during high CO2 concentrations might be a chance finding. Hence, replication is needed in actual bedrooms controlling for other external factors before any generalizations can be made. Int J Occup Med Environ Health. 2023;36(2):177-91.

Epigenetic Silencing of LRP2 Is Associated with Dedifferentiation and Poor Survival in Multiple Solid Tumor Types.

More than 80% of human cancers originate in epithelial tissues. Loss of epithelial cell characteristics are hallmarks of tumor development. Receptor-mediated endocytosis is a key function of absorptive epithelial cells with importance for cellular and organismal homeostasis. LRP2 (megalin) is the largest known endocytic membrane receptor and is essential for endocytosis of various ligands in specialized epithelia, including the proximal tubules of the kidney, the thyroid gland, and breast glandular epithelium. However, the role and regulation of LRP2 in cancers that arise from these tissues has not been delineated. Here, we examined the expression of LRP2 across 33 cancer types in The Cancer Genome Atlas. As expected, the highest levels of LRP2 were found in cancer types that arise from LRP2-expressing absorptive epithelial cells. However, in a subset of tumors from these cancer types, we observed epigenetic silencing of LRP2. LRP2 expression showed a strong inverse correlation to methylation of a specific CpG site (cg02361027) in the first intron of the LRP2 gene. Interestingly, low expression of LRP2 was associated with poor patient outcome in clear cell renal cell carcinoma, papillary renal cell carcinoma, mesothelioma, papillary thyroid carcinoma, and invasive breast carcinoma. Furthermore, loss of LRP2 expression was associated with dedifferentiated histological and molecular subtypes of these cancers. These observations now motivate further studies on the functional role of LRP2 in tumors of epithelial origin and the potential use of LRP2 as a cancer biomarker.