Environmental and genetic drivers of population differences in SARS-CoV-2 immune responses

Humans display vast clinical variability upon SARS-CoV-2 infection1-3, partly due to genetic and immunological factors4. However, the magnitude of population differences in immune responses to SARS-CoV-2 and the mechanisms underlying such variation remain unknown. Here we report single-cell RNA-sequencing data for peripheral blood mononuclear cells from 222 healthy donors of various ancestries stimulated with SARS-CoV-2 or influenza A virus. We show that SARS-CoV-2 induces a weaker, but more heterogeneous interferon-stimulated gene activity than influenza A virus, and a unique pro-inflammatory signature in myeloid cells. We observe marked population differences in transcriptional responses to viral exposure that reflect environmentally induced cellular heterogeneity, as illustrated by higher rates of cytomegalovirus infection, affecting lymphoid cells, in African-descent individuals. Expression quantitative trait loci and mediation analyses reveal a broad effect of cell proportions on population differences in immune responses, with genetic variants having a narrower but stronger effect on specific loci. Additionally, natural selection has increased immune response differentiation across populations, particularly for variants associated with SARS-CoV-2 responses in East Asians. We document the cellular and molecular mechanisms through which Neanderthal introgression has altered immune functions, such as its impact on the myeloid response in Europeans. Finally, colocalization analyses reveal an overlap between the genetic architecture of immune responses to SARS-CoV-2 and COVID-19 severity. Collectively, these findings suggest that adaptive evolution targeting immunity has also contributed to current disparities in COVID-19 risk.

Cannabis for pain: a cross-sectional survey of the patient information quality on the Internet.

BACKGROUND: Cannabis has increasingly become an alternative treatment for chronic pain, however, there is evidence of concomitant negative health effects with its long-term usage. Patients contemplating cannabis use for pain relief commonly see information online but may not be able to identify trustworthy and accurate sources, therefore, it is imperative that healthcare practitioners play a role in assisting them in discerning the quality of information. The present study assesses the quality of web-based consumer health information available at the intersection of cannabis and pain. METHODS: A cross-sectional quality assessment of website information was conducted. Three countries were searched on Google: Canada, the Netherlands, and the USA. The first 3 pages of generated websites were used in each of the 9 searches. Eligible websites contained cannabis consumer health information for pain treatment. Only English-language websites were included. Encyclopedias (i.e. Wikipedia), forums, academic journals, general news websites, major e-commerce websites, websites not publicly available, books, and video platforms were excluded. Information presented on eligible websites were assessed using the DISCERN instrument. The DISCERN instrument consists of three sections, the first focusing on the reliability of the publication, the second investigating individual aspects of the publication, and the third providing an overall averaged score. RESULTS: Of 270 websites identified across searches, 216 were duplicates, and 18 were excluded based on eligibility criteria, resulting in 36 eligible websites. The average summed DISCERN score was 48.85 out of 75.00 (SD = 8.13), and the average overall score (question 16) was 3.10 out of 5.00 (SD = 0.62). These overall scores were calculated from combining the scores for questions 1 through 15 in the DISCERN instrument for each website. Websites selling cannabis products/services scored the lowest, while health portals scored the highest. CONCLUSION: These findings indicate that online cannabis consumer health information for the treatment/management of pain presents biases to readers. These biases included websites: (1) selectively citing studies that supported the benefits associated with cannabis use, while neglecting to mention those discussing its risks, and (2) promoting cannabis as "natural" with the implication that this equated to "safe". Healthcare providers should be involved in the guidance of patients' seeking and use of online information on this topic.

Myalgic encephalomyelitis: International Consensus Criteria.

The label 'chronic fatigue syndrome' (CFS) has persisted for many years because of the lack of knowledge of the aetiological agents and the disease process. In view of more recent research and clinical experience that strongly point to widespread inflammation and multisystemic neuropathology, it is more appropriate and correct to use the term 'myalgic encephalomyelitis' (ME) because it indicates an underlying pathophysiology. It is also consistent with the neurological classification of ME in the World Health Organization's International Classification of Diseases (ICD G93.3). Consequently, an International Consensus Panel consisting of clinicians, researchers, teaching faculty and an independent patient advocate was formed with the purpose of developing criteria based on current knowledge. Thirteen countries and a wide range of specialties were represented. Collectively, members have approximately 400 years of both clinical and teaching experience, authored hundreds of peer-reviewed publications, diagnosed or treated approximately 50 000 patients with ME, and several members coauthored previous criteria. The expertise and experience of the panel members as well as PubMed and other medical sources were utilized in a progression of suggestions/drafts/reviews/revisions. The authors, free of any sponsoring organization, achieved 100% consensus through a Delphi-type process. The scope of this paper is limited to criteria of ME and their application. Accordingly, the criteria reflect the complex symptomatology. Operational notes enhance clarity and specificity by providing guidance in the expression and interpretation of symptoms. Clinical and research application guidelines promote optimal recognition of ME by primary physicians and other healthcare providers, improve the consistency of diagnoses in adult and paediatric patients internationally and facilitate clearer identification of patients for research studies.

Tailored adsorption of His6-tagged protein onto nickel(II)-cyclam grafted mesoporous silica.

The specific immobilisation of a histidine-tagged protein, Spi, onto mono-anchored nickel(II)-cyclam functionalised SBA-15 is reported.

Mineral retention in three-week-old piglets fed goat and cow milk infant formulas.

Goat milk and cow milk are commonly used in infant formula preparations and, as such, understanding the nutritional characteristics of infant formulas made from these milks is important. In this study, a goat milk infant formula was compared with an adapted (whey-enhanced) cow milk infant formula with respect to mineral absorption and deposition using the 3-wk-old piglet as a model for the 3-mo-old infant. Equal numbers of piglets (n = 8) were fed either the goat milk formula or the cow milk formula. The mineral composition of the prepared goat milk formula was higher than that of the prepared cow milk formula for most minerals, including calcium (75.1 vs. 56.7 mg/100 mL) but excluding iron, which was higher in the prepared cow milk formula (0.92 vs. 0.74 mg/100 mL). The amounts of calcium, phosphorus, and manganese absorbed by the piglets were significantly higher for the goat milk formula, whereas the amounts of zinc, iron, and magnesium absorbed were significantly higher for the cow milk formula. Apparent mineral absorption, relative to intake, was statistically higher in the cow milk formula for calcium and phosphorus, although the actual differences were very small (less than 1.3%). For copper, zinc, iron, and magnesium there was no significant difference between treatments in apparent mineral absorption, whereas for manganese, absorption was higher for the goat milk infant formula. The absolute mineral deposition was higher in piglets fed the goat milk formula for calcium, phosphorus, and manganese, whereas iron deposition was higher in the piglets fed cow milk formula. For all other minerals tested, there were no significant differences between treatments. The goat milk infant formula provided a pattern of mineral retention in the 3-wk-old piglet very similar to that of the adapted cow milk infant formula. The minor differences observed between the 2 appeared to be due to the different mineral contents of the 2 formulas.

True ileal amino acid digestibility of goat and cow milk infant formulas.

Goat milk is used as an alternative to cow milk for the production of infant formulas. However, little is known about the protein quality and, specifically, about the digestible AA pattern of goat milk formulas compared with their cow milk counterparts. In this study, the true ileal AA digestibility of a goat milk infant formula was compared with a premium cow milk infant formula. The 3-wk-old piglet was used as a model for the 3-mo-old infant. Both milk formulas were prepared as described by the manufacturer, with titanium dioxide added as an indigestible marker. The formulas were fed to the piglets over a 2-wk trial period. Digesta from the terminal ileum were collected post euthanasia and analyzed for AA content, along with samples of the formulas. True AA digestibility was determined after correcting for endogenous AA loss at the terminal ileum of pigs fed an enzyme-hydrolyzed casein-based diet, followed by ultrafiltration (5,000 Da) of the digesta. Total urine and feces collection was also undertaken to determine the nitrogen retention from the diets. The true ileal AA digestibility was similar between the goat and cow milk infant formulas for all AA except Gly and Trp. There was no significant difference in the nitrogen retention of piglets fed the two different formulas. The goat milk infant formula and the premium cow milk infant formula were similar in terms of protein quality.

Ascertainment of pesticide exposures of migrant and seasonal farmworker children: findings from focus groups.

BACKGROUND: To design questionnaires for epidemiologic research among children of migrant farmworkers, researchers need to consider ways to best solicit information about pesticide exposures. METHODS: Bilingual facilitators conducted five focus groups with either migrant farmworker mothers or their children (age range 8-16 years) in southern Texas and northeastern Colorado. Guided questions were used to assess activities of migrant farmworker children and the ways to best elicit information about exposure to pesticides. RESULTS: Participants reported a large number of activities that may potentially expose children to pesticides through both direct and indirect routes. Prompting, indirect questions about chemical use, and use of local and trusted facilitators increased information elicited from focus group participants. CONCLUSIONS: These focus groups helped to provide information for developing questionnaire items related to pesticide exposure among migrant farmworker children, and highlighted the importance of using bilingual community interviewers and including children as respondents.

Optimizing immunity and gut function in the elderly.

The elderly represent a group for whom health issues are of paramount concern. Physiological processes, including the functioning of the immune and digestive systems, are known to decline with age, and consequently aging can increase susceptibility to infectious and non-infectious disease. Dietary supplementation may offer a safe and effective means of countering physiological senescence, by optimizing important immune and gut functions. This review outlines our current understanding of the ways in which aging affects immune and gut function, and discusses clinical evidence for a role of dietary supplementation in countering age-related deficiencies in these processes.

"Tears in my eyes 'cause somebody finally understood": client perceptions of practitioners following brain injury.

OBJECTIVE: The purpose of this study was to identify practitioner qualities and traits that clients with brain injury see as important. METHOD: An opportunistic sample of 51 participants with brain injury was interviewed about perceptions of service access and effectiveness. An interview guide was used to gather data for this phenomenological qualitative study. Four interviewers individually conducted audiotaped interviews, which were then transcribed. Coding and theme development were completed using HyperRESEARCH software. RESULTS: Three themes regarding practitioner qualities emerged from the data: (a) roles of the provider, (b) perceived helpfulness of services, and (c) personal characteristics of the providers. Beneficial provider roles included advocate, friend, mentor, and team member. Perceptions of helpfulness of the services included relevance, meaningfulness, practical application, skill development potential, and whether periodic feedback on progress was provided. Personal characteristics of the provider valued by the participants were clear and honest communicator, supportive, respectful, good listener, and understanding. CONCLUSIONS: Practitioners need to pay increased attention to the perceptions of care and services of clients with brain injury. The current study closely supports previous research related to quality of care in the medical and community arenas and offers some additional suggestions to professionals who work with persons with brain injury, including learning how to time giving information to clients and how to understand client concerns without being prescriptive. Future research in this area needs to focus on and describe providers who demonstrate an ability-through their willingness to don a variety of roles, their helpful services, and their personal characteristics-to meet the unique needs of clients with brain injury.

Quantifying the digestibility of dietary protein.

The current recommendation, when calculating a protein digestibility-corrected amino acid score, is to determine the digestibility of a dietary protein across the entire digestive tract, using the rat as a model animal for humans. This fecal digestibility value is subsequently corrected for endogenous contributions of protein using a metabolic nitrogen value determined by feeding rats a protein-free diet. The limitations inherent with this method are well recognized, however, and determining the digestibility of a dietary protein to the end of the small intestine is the preferred alternative. Unlike the fecal digestibility assay, which has only one basic methodology, ileal digestibility values can be determined in a number of ways. We discuss the various methods available for determining ileal digestibility values and compare results obtained for dietary proteins using both fecal and ileal digestibility assays. The relative value of using individual amino acid digestibility values as opposed to nitrogen digestibility values is reviewed. In addition, we address issues surrounding measurement of endogenous nitrogen flows, and in particular, the relative merits of determining "true" versus "real" digestibility values.

The amino acid composition of human milk corrected for amino acid digestibility.

Human milk was collected from women in their 10th-14th weeks of lactation, and was analysed for amino acids. Corrections were made for losses of amino acids which were presumed to occur during acid hydrolysis, using a non-linear mathematical model that describes the simultaneous processes of amino acid yield and decay. The mean amino acid composition of the human milk was found to be similar to previously reported estimates, although the cysteine content of the human milk in the present study was 20% higher than the average literature estimate. True (corrected for endogenous amino acid excretions) ileal amino acid digestibility of human milk was determined using the 3-week-old piglet as a model animal for the human infant. The piglets were given either human milk (n 6) or a protein-free diet (n 6) for a 6 d experimental period. Cr2O3 was added as an indigestible marker, to both the human milk and protein-free diet. At the end of the experimental period the piglets were anaesthetized and samples of digesta removed from the terminal ileum of each piglet. After sampling the piglets were killed. Endogenous ileal excretions of amino acids were determined in piglets fed on the protein-free diet. The true digestibilities of total N and amino acid N were 88% and 95% respectively. The true ileal digestibility of the non-amino acid N fraction in human milk, when calculated by difference was only 50%. The true digestibility of the amino acids in human milk ranged from 81-101% with threonine (86%) being the least digestible essential amino acid. When the true ileal digestibility values were used to correct the amino acid composition of human milk, the pattern of digestible amino acids in human milk was different compared with the currently recommended pattern of amino acid requirements for the infant.

Perceptions of care access: the experience of rural and urban women following brain injury.

To examine care access among women with brain injuries, a qualitative, phenomenological study was conducted. Twenty-one women with acquired brain injuries were recruited for the inquiry, representing rural and urban regions of Colorado. The phenomenological approach allowed the investigators to examine the shared experiences of women with brain injuries as they sought rehabilitative supports and services, and attempted to re-enter their lives and communities. Each woman was interviewed one or two times, the interviews were transcribed and then coded and analysed by both authors separately. Thematic analysis of the data revealed two major emergent themes: issues surrounding diagnosis and barriers to accessing care. A total of 10 women interviewed experienced difficulty obtaining a diagnosis of brain injury and experienced serious personal and professional consequences as a result. All the women experienced barriers to accessing care, including problems with service providers and service systems, financial challenges, travelling for services, lack of information or services in their area, lack of care coordination and funneling. Some of the barriers cannot be changed secondary to cost, efficiency and logistics. Other barriers, could be changed through education, training and policy making.

Environment effect of functional task performance in adults with acquired brain injuries: use of the assessment of motor and process skills.

OBJECTIVE: To examine household task performance in both familiar (home) and unfamiliar (clinic) environments in adults with acquired brain injuries. DESIGN: The research performed was a comparison study examining the effect of the environment on functional task performance. Individuals were evaluated using the Assessment of Motor and Process Skills both in their homes and in an unfamiliar clinic setting. PARTICIPANTS: Twenty individuals with acquired brain injuries living in the community. MAIN OUTCOME MEASURES: The Assessment of Motor and Process Skills, an observational, standardized, occupational therapy assessment tool, was used to evaluate household task performance. RESULTS: Paired one-tailed t tests indicated a significant difference between home and clinic performance measures in process ability (t=-4.28, p=.00), but no significant difference in motor ability performance measures (t=-1.84, p=.410). Scatter plot analyses showed that performance scores of 6 of the 20 subjects differed in a clinically meaningful way, and that instrumental activities of daily living motor performance scores for 3 of the 20 differed in a clinically meaningful way. CONCLUSIONS: Individuals with acquired brain injuries may be influenced by their environment when performing household tasks, although further study is necessary to determine the extent of the environmental effect.

Pharmacokinetics and pharmacodynamics of recombinant human interferon-beta in healthy male volunteers.

The pharmacokinetics and pharmacodynamics of recombinant human interferon-beta (rHuIFN-beta 1a) were assessed following administration to 12 healthy male volunteers. Each subject received, in a double-blind, balanced, random-order, crossover sequence, single doses of 6 MIU of rHuIFN-beta 1a (Rebif) i.v., i.m., and s.c. or matching placebo on four occasions separated by washout periods of 1 week. Blood samples were collected at preset times for the measurement of serum IFN-beta levels and of intracellular 2'-5'-oligoadenylate synthetase levels. Blood pressure, sitting heart rate, respiratory rate, oral body temperature, and tolerance were monitored regularly. All administrations of rHuIFN-beta 1a were well tolerated, although about half of the subjects had a flu-like syndrome, as expected. After i.v. bolus injection, the pharmacokinetics of rHuIFN-beta 1a were well described by a classic two-compartment model. Mean total clearance of rHuIFN-beta 1a was about 100 L.h-1. The distribution half-life was 5 min, and the terminal half-life was approximately 5 h. After i.m. or s.c. injection, serum IFN-beta profiles were rather flat, and about one sixth of the administered dose was available systemically. Extent and duration of clinical and biologic effects were independent of the route of administration and of the IFN-beta serum levels. Biologic pharmacodynamic effects persisted even when IFN-beta serum levels had returned to baseline and were still significantly elevated 3 days after a single dose. Because of the independence of the extent and duration of clinical and biologic pharmacodynamic effects from the route of administration and from the IFN-beta serum levels, the s.c route of administration is preferred in indications in which primarily an immunomodulatory action is sought. Predominantly antiviral and antiproliferative activity is enhanced by the i.v. route to provide adequate drug levels at the site of pathology, although its application is limited on practical grounds.

Correction for amino acid loss during acid hydrolysis of a purified protein.

Hydrolyzing a protein in acid for a single hydrolysis interval, normally 24 h, will lead to inaccurate estimates of the amino acid composition of that protein due to an effect of the time of hydrolysis on peptide bond cleavage and amino acid degradation. The simultaneous yield and decay of amino acids during the hydrolysis of a protein can be described by a compartmental model with parameters for the hydrolysis and loss rates specific to each amino acid in a protein. The amino acid composition of the protein prior to hydrolysis can be determined by nonlinear regression of data derived from multiple hydrolysis intervals. In the present study egg-white lysozyme was hydrolyzed in 6 M HCl using 18 hydrolysis intervals (range, 2-141 h) using the conventional duplicate hydrolyses/interval system. Hydrolysis and loss rates were determined for each amino acid. Increasing the number of hydrolysis intervals prior to the maximum point on the hydrolysis curve, and including an hydrolysis interval greater than 100 h increased the accuracy with which the hydrolysis and loss rates were estimated. Most of the amino acids underwent some degree of loss during hydrolysis. Of particular note was the loss rate for cysteic acid, which was greater than that found for serine which is commonly regarded as an acid-labile amino acid. The determined amino acid composition of the protein, based on the nonlinear regression of the data from four different series of hydrolysis intervals, was compared with the known amino acid composition (sequencing). Using the routine duplicate sampling system, a nonlinear regression including 10 hydrolysis intervals (2, 6, 10, 14, 18, 22, 26, 30, 60, and 141 h) resulted in a mean amino acid recovery of 100% (range, 94-110%) and provided an acceptable compromise between accuracy and the cost of analysis.

The three-week-old piglet as a model animal for studying protein digestion in human infants.

The piglet was evaluated as a model animal for studying the digestion of high-quality proteins in human infants. Three-week-old male piglets (n = 6) and 3-month-old male human infants (n = 6) were fed a bovine-milk-based formula over a 17-day experimental period comprising 7 days' adaptation followed by a 10-day fecal collection period. The piglets and infants were given 345-g liquid formula/kg body weight/day and 170-g liquid formula/kg body weight/day, respectively, which equated to similar dry matter intakes per unit stomach volume (0.923 g dry matter/cm3/day). Both the piglets and infants were individually bottle-fed the reconstituted milk formula (12.2% dry matter) at similar meal frequencies. Small but statistically significant differences (p < 0.01) were found for the apparent fecal digestibility (mean +/- overall SE) of dietary dry matter (98.8 versus 97.4% +/- 0.13%), organic matter (99.0 versus 97.7% +/- 0.12%), and total nitrogen (97.5 versus 94.5% +/- 0.36%) between the piglets and infants. The fecal digestibilities for most of the amino acids were not significantly different (p > 0.05) between the species. The digestion of protein appeared to be similar in the two species. The study provides support for using the piglet as a model animal for studying protein digestion in human infants.

Absorption of lysine and methionine from the proximal colon of the piglet.

The present study aimed to determine whether lysine and/or methionine are absorbed in nutritionally significant amounts from the proximal colon of milk-formula-fed piglets (15-32 d old; 2.0-7.4 kg liveweight). Piglets, surgically prepared with simple catheters which allowed infusion into the proximal colon, were randomly allocated to one of two milk-formula diets which were either 40% deficient in lysine (L-diet) or 60% deficient in methionine and 40% deficient in cysteine (S-diet), yet balanced for all other amino acids. The piglets were individually bottle-fed the milk-formula diets seven times daily at 2 h intervals between 08.00 and 20.00 hours. Physiological saline (9 g NaCl/l) or an isotonic solution containing the deficient amino acid was infused via the catheter at each feeding. The experimental procedure followed a cross-over design. Total daily excretions of urinary urea and total N were determined. There were no significant differences (P > 0.05) in urinary N metabolite excretion for piglets infused with amino acids compared with those infused with saline. Lysine and methionine do not appear to be absorbed in nutritionally significant amounts from the proximal colon of the milk-fed piglet.

The effect of age on the pharmacokinetics of pentisomide.

The effects of age on the pharmacokinetics of pentisomide (CM7857), an orally effective antiarrhythmic agent, were studied in two groups of volunteers. Sixteen young volunteers (mean age 26.4 years) and 10 elderly volunteers (mean age 67.8 years) received a single 200 mg oral dose of pentisomide. Mean AUC was larger and terminal elimination half-life longer in the elderly subjects, due to a decrease in total plasma clearance of pentisomide in the elderly. This decrease was due to a reduction in renal clearance of the drug which was paralleled by a significantly lower creatinine clearance in the elderly subjects. Dosage reduction, or a reduced frequency of dosing of pentisomide would be necessary in the elderly or those with impaired renal function.

Pharmacokinetics of the anti-inflammatory drug ximoprofen in healthy young and elderly subjects: comparison with elderly rheumatic patients.

The pharmacokinetics of ximoprofen were studied in young and elderly subjects after single and repeated doses up to 30 mg. In healthy elderly subjects (30 mg dose), a mean peak plasma drug concentration of 1.78 micrograms ml-1 +/- 0.83 s.d. occurred at a mean time of 1.95 h +/- 1.40 s.d. and, thereafter, concentrations declined monoexponentially with a mean half-life of 3.8 h +/- 1.4 s.d. Comparison of these data with those from younger healthy subjects showed that peak drug concentrations, areas under the curve and half-lives were about two-fold greater in the elderly, these differences probably reflecting a lower systemic drug clearance. Similar results were obtained on comparing data from young healthy subjects and elderly rheumatic patients receiving single and repeated doses of ximoprofen (15 mg twice daily). In patients, the half-life of ximoprofen was 2.5 h +/- 0.7 s.d. Within either group, pharmacokinetic parameters after single or repeated doses were similar: ximoprofen did not accumulate in the plasma of the young or elderly.

Pharmacokinetics of the anti-inflammatory drug ximoprofen in healthy subjects and in disease states.

The pharmacokinetics of ximoprofen, a potent new non-steroidal anti-inflammatory agent, has been investigated in normal healthy subjects and in patients with hepatic or renal disease. After intravenous infusion of 22.8 mg to healthy subjects, plasma ximoprofen concentrations declined in a polyexponential manner with a terminal phase half-life of 1.9 h. The systemic clearance of ximoprofen was 115 ml.min-1 and the volumes of distribution were 18.01 Vz and 13.81 Vss. Ximoprofen was 80-90% bound to plasma proteins. The systemic availabilities (f) of orally and rectally administered doses of 30 mg of ximoprofen were 98% and 56% respectively and, in the case of the rectal dose, absorption appeared to be prolonged leading to "flip-flop" kinetics. After single oral doses of 30 mg of ximoprofen to patients with hepatic disease, half-life (2.2 h), peak plasma concentrations (1.55 micrograms.ml-1 cf 1.04 micrograms.ml-1 in healthy subjects) and areas under the curve (6.12 micrograms.h.ml-1 cf 3.54 micrograms.h.ml-1 in healthy subjects) were significantly different from those in healthy subjects. After single oral doses of 30 mg of ximoprofen to patients with renal disease, pharmacokinetic parameters of half-life (4.0 h), mean residence time (6.0 h) and area under the curve (9.2 micrograms.h.ml-1) were significantly different from those in healthy subjects. There were no significant differences in pharmacokinetic parameters between patients having differing degrees of renal disease. These data nevertheless suggest that accumulation of ximoprofen in hepatic or renal disease would be of slight or negligible clinical relevance and that no alteration of the dose regimen (up to 15 mg twice daily) may be required when ximoprofen is administered in these disease states.