Pulmonary inflammation promoted by type-2 dendritic cells is a feature of human and murine schistosomiasis.

Schistosomiasis is a parasitic disease affecting over 200 million people in multiple organs, including the lungs. Despite this, there is little understanding of pulmonary immune responses during schistosomiasis. Here, we show type-2 dominated lung immune responses in both patent (egg producing) and pre-patent (larval lung migration) murine Schistosoma mansoni (S. mansoni) infection. Human pre-patent S. mansoni infection pulmonary (sputum) samples revealed a mixed type-1/type-2 inflammatory cytokine profile, whilst a case-control study showed no significant pulmonary cytokine changes in endemic patent infection. However, schistosomiasis induced expansion of pulmonary type-2 conventional dendritic cells (cDC2s) in human and murine hosts, at both infection stages. Further, cDC2s were required for type-2 pulmonary inflammation in murine pre-patent or patent infection. These data elevate our fundamental understanding of pulmonary immune responses during schistosomiasis, which may be important for future vaccine design, as well as for understanding links between schistosomiasis and other lung diseases.

Dynamics of the bacterial gut microbiota during controlled human infection with Necator americanus larvae.

Hookworms are soil-transmitted helminths that use immune-evasive strategies to persist in the human duodenum where they are responsible for anemia and protein loss. Given their location and immune regulatory effects, hookworms likely impact the bacterial microbiota. However, microbiota studies struggle to deconvolute the effect of hookworms from confounders such as coinfections and malnutrition. We thus used an experimental human hookworm infection model to explore temporal changes in the gut microbiota before and during hookworm infection. Volunteers were dermally exposed to cumulative dosages of 50, 100 or 150 L3 Necator americanus larvae. Fecal samples were collected for microbiota profiling through 16S rRNA gene amplicon sequencing at weeks zero, four, eight, fourteen and twenty. During the acute infection phase (trial week zero to eight) no changes in bacterial diversity were detected. During the established infection phase (trial week eight to twenty), bacterial richness (Chao1, p = .0174) increased significantly over all volunteers. No relation was found between larval dosage and diversity, stability or relative abundance of individual bacterial taxa. GI symptoms were associated with an unstable microbiota during the first eight weeks and rapid recovery at week twenty. Barnesiella, amongst other taxa, was more abundant in volunteers with more GI symptoms throughout the study. In conclusion, this study showed that clinical GI symptoms following N. americanus infection are associated with temporary microbiota instability and relative abundance of specific bacterial taxa. These results suggest a possible role of hookworm-induced enteritis on microbiota stability.

Physiological stress response patterns during a blood donation.

BACKGROUND: Donating blood is associated with increased psychological stress. This study investigates whether a blood donation induces physiological stress and if response patterns differ by gender, donation experience and non-acute stress. STUDY DESIGN AND METHODS: In 372 donors, physiological stress [blood pressure, pulse rate, pulse rate variability (PRV)] was measured at seven moments during routine donation. PRV was assessed using time domain [root mean square of successive differences (RMSSD)] and frequency domain [high frequency (HF) and low frequency (LF) power] parameters. Non-acute stress was assessed by questionnaire. Shape and significance of time course patterns were assessed by fitting multilevel models for each stress measure and comparing men and women, first-time and experienced donors, and donors with high and low levels of non-acute stress. RESULTS: Significant response patterns were found for all stress measures, where levels of systolic blood pressure (F(1,1315) = 24·2, P < 0·001), RMSSD (F(1,1315) = 24·2, P < 0·001), LF (F(1,1627) = 14·1, P < 0·001) and HF (F(1,1624) = 34·0, P < 0·001) increased towards needle insertion and then decreased to values lower than when arriving at the donation centre. Diastolic blood pressure (F(1,1326) = 50·9, P < 0·001) increased and pulse rate (F(1,1393) = 507·4, P < 0·001) showed a U-shaped curve. Significant group effects were found, that is, higher systolic blood pressure/pulse rate in women; higher pulse rate in first-time donors; higher RMSSD at arrival and from screening until leaving in first-time donors; and higher LF and HF in first-time donors. CONCLUSION: This study shows an increase in physiological stress related to needle insertion, followed by a decrease when leaving the donation centre. Some group effects were also found.

Psychological and hormonal stress response patterns during a blood donation.

BACKGROUND AND OBJECTIVES: Donating blood has been associated with increased stress responses, with scarce evidence indicating that levels of psychological and hormonal stress are higher pre-donation than post-donation. We investigated whether a blood donation induces psychological and/or hormonal stress during the course of a blood donation, and whether responses differed between men and women, first-time and experienced donors and donors with high or low non-acute stress. MATERIALS AND METHODS: In 363 donors, psychological (donation-stress and arousal) and hormonal (cortisol) stress were measured by questionnaire and salivary sample at seven key moments during a routine donation. Non-acute stress was assessed by a questionnaire. Repeated measurement analyses were performed, using the last measurement (leaving the donation center) as reference value. RESULTS: Levels of donation-stress, arousal and cortisol were significantly higher during donation than when leaving the donation center. When compared with men, women reported higher levels of donation-stress and cortisol in the first part of the visit. When compared with first-time donors, experienced donors reported lower levels of donation-stress during the first part of the visit, and higher levels of arousal but less reactivity throughout the visit. When compared to donors high on non-acute stress, donors low on non-acute stress reported lower levels of donation-stress during the first part of the visit, and showed less cortisol reactivity throughout the visit. CONCLUSION: Donating blood influences psychological and hormonal stress response patterns. The response patterns differ between women and men, first-time and experienced donors and between donors high and low on non-acute stress.

Negative experiences and pre-donation blood pressure: the role of attitude and anxiety.

BACKGROUND/OBJECTIVES: Negative experiences (NEs) have been shown to result in an increased stress response, as indicated by blood pressure, at the subsequent donation. This response might be influenced by how the donor rates the donation in terms of importance and pleasantness [affective attitude (AA)/cognitive attitude (CA)] or by anxiety about donating blood. We investigated the effect of AA/CA/anxiety on the impact of NEs on pre-donation blood pressure (pd-BP) in the subsequent donation. MATERIALS/METHODS: pd-BP at visit 3 was compared between donors with and without a history of NEs during or after their first two visits (visit 1: medical check, visit 2: first donation). The effect of AA/CA/anxiety (measured 1 month prior to visit 1 on a 7-point scale) on visit 3 pd-BP was explored using linear regression and interaction analyses. Analyses were stratified for gender, age and pd-BP at visit 1, which were taken into account as confounders. RESULTS: In 1106 first-time blood donors (70% female), 632 donors (57% of total) indicated an NE at their first donation. Mean scores for AA/CA/anxiety were 5·2/6·5/2·2 (men without NE), 4·8/6·3/3·0 (men with NE), 5·2/6·6/2·6 (women without NE) and 4·8/6·6/3·2 (women with NE). No significant associations were found for NE and pd-BP at visit 3 after adjusting for confounding. Of 48 interaction effects, four were significant, but effects were small and inconsistent. CONCLUSION: In donors who had had negative experiences during their first donation, anxiety and attitude to donation did not influence their pre-donation blood pressure at their subsequent visit.

Negative experiences and predonation blood pressure at the subsequent donation in blood donors.

BACKGROUND: Negative donation experiences, like being deferred or experiencing an adverse reaction, might upset blood donors, resulting in anticipatory stress responses such as elevated blood pressure at the subsequent visit. We therefore explored associations between blood donors' negative donation experiences and their blood pressure at the subsequent visit. STUDY DESIGN AND METHODS: Blood pressure of donors with no history of negative experiences in three consecutive donations was compared to the blood pressure of donors with a negative experience during the second of the three donations. Blood pressure (systolic and diastolic) measured prior to the third donation was compared between the two groups, using linear regression analyses. Two types of negative experiences (adverse reactions and deferral) were analysed, stratifying for donation type and sex, and adjusting for age and predonation blood pressure at baseline. RESULTS: In total, 248 118 (50% female) donors were included in the analyses. Eleven per cent (26 380 donors, 61% female) had experienced a negative experience. Fainting and dizziness were associated with significant (P < 0·05) increases in systolic blood pressure: in men, 3·0 mmHg (fainting) and 2·0 mmHg (dizziness); in women, 2·0 mmHg (fainting) and 1·4 mmHg (dizziness). Deferral was associated with significant (P < 0·05) increases in both systolic (men: 0·7 mmHg, women: 0·3 mmHg) and diastolic (men: 0·2 mmHg, women: 0·3 mmHg) blood pressure. CONCLUSION: Whole blood donations with negative experiences were associated with a statistically significant higher predonation blood pressure at the subsequent visit. This indicates that negative experiences might cause an anticipatory stress reaction in a subsequent donation.

"Feelings are facts": illness perceptions in patients with lung cancer.

Given the high degree of psychosocial problems in patients with lung cancer, quality medical care would benefit from exploring and addressing and providing potential solutions for these problems. Patients with recently diagnosed non-small-cell lung cancer filled out a questionnaire that assessed illness perceptions and made a drawing of how they perceived their diseased lungs look. They also participated in an interview about the impact of lung cancer in their lives. Scores on the Brief Illness Perception Questionnaire indicated that patients score low on 'concern', 'emotional response' and 'timeline', indicating they hope to be cured from lung cancer. Patients drew the tumor larger than it is on the chest radiograph. The drawings are moderately accurate representations of the patients' lungs. In the interviews patients often expressed their hopes of being cured and how thinking positively would help. Patients who made a more accurate drawing of their lungs had less optimistic views about their prognosis. These views are more in line with the prognosis their physician would give them. However, few patients made an accurate drawing. This study contributes to a better insight into what patients believe and feel about their disease. Suggestions for taking patient perceptions into account are provided.

Nutritional counselling for diabetic renal patients.

A diabetic patient who develops nephropathy has to cope with two different diets. Healthy food guidelines have to be followed and attention paid to the spread of carbohydrates throughout the day because of the diabetes. Renal failure means that attention also has to be paid to protein, sodium, potassium and fluid intake and the diet can be confusing to patients. When dialysis commences the management of, and attention to the diabetes is sometimes a little forgotten. This article identifies problems and how they may be dealt with.

Nutritional aspects of acute renal failure.

The patient with acute renal failure is a very ill patient suffering from high urea levels causing poor appetite, nausea and vomiting. These patients are usually treated with a low sodium, low protein and, if the potassium in the blood is high, with a low potassium diet (1). This paper discusses whether or not this is the correct treatment. The symptoms of high urea levels in the blood together with increased needs for energy and protein makes it very hard to prevent the patient becoming malnourished. Often energy-enriched drinks are necessary to achieve recommendations and it is prudent to let the patient eat and drink what they desire. By calculating the energy and protein needs and comparing these with the intake and the state of illness and by following the patient's body weight over time we can obtain information about the state of nourishment. When we alter the food that's offered we achieve better intake and reduce the risk of malnourishment.

Cross-linking of the carcinoembryonic antigen-like glycoproteins CD66 and CD67 induces neutrophil aggregation.

The carcinoembryonic Ag (CEA)-like glycoproteins present on human granulocytes have been designated non-specific cross-reacting Ag (NCA). We have recently demonstrated that granulocyte-specific CD66 and CD67 mAb recognize the three originally described NCA. CD66 binds to NCA-160 and NCA-90, whereas CD67 only recognizes NCA-95. As we have shown previously, NCA-160 and NCA-90 present sialylated Lewis-X oligosaccharide Ag (SLex) in a functional way, i.e., these Ag function as (one of many possible) molecules involved in neutrophil binding to the adhesion molecule E-selectin expressed on activated endothelial cells. In this study, we found that a polyclonal anti-CEA antiserum, either as intact Ig or as F(ab')2 fragments, induced neutrophil aggregation. This aggregation response was blocked by CD18 mAb. Neutrophils from a patient severely affected by paroxysmal nocturnal hemoglobinuria completely lacked expression of NCA-95 and NCA-90. The patient's neutrophils repeatedly showed no aggregation on addition of the anti-CEA antiserum. Thus, the presence of the phosphatidylinositol-linked NCA-95/90 seems to be essential for NCA-induced neutrophil activation. Our data indicate that NCA molecules on neutrophils may function via binding to E-selection or an as yet unknown ligand (as mimicked here by an antiserum) and subsequently induce CD18-dependent adhesive properties.

Cross-linking of sialophorin (CD43) induces neutrophil aggregation in a CD18-dependent and a CD18-independent way.

Normal human neutrophils bound an as yet unclustered mAb designated BS-1. The Ag immunoprecipitated with BS-1 was blotted by CD43 mAb (and vice versa), and is therefore identical to the large sialoglycoprotein. The CD43 Ag expression on the neutrophil surface is decreased upon neutrophil activation with the chemoattractant FMLP or with PMA. This can be (at least partially) explained by the release of CD43+ material with an altered electrophoretic mobility into the extracellular medium of the neutrophils upon activation. Cross-linking of the CD43 Ag with BS-1 also invoked neutrophil activation by itself: F(ab)2 fragments of BS-1-induced neutrophil aggregation, in contrast to F(ab) fragments. Neither respiratory burst activity nor a significant rise in intracellular Ca2+ level or actin polymerization were observed. The transient neutrophil aggregation response was largely CD18 dependent, especially in the initial phase of homotypic clustering. However, a significant CD18-independent mechanism contributed thereafter to the neutrophil aggregation, as was further substantiated by the use of cultured T (and EBV-transformed B) cell clones of a patient with a leukocyte adhesion deficiency. CD43 is the first molecule described on neutrophils able to induce adhesive properties in a dual fashion.

CD66 nonspecific cross-reacting antigens are involved in neutrophil adherence to cytokine-activated endothelial cells.

Neutrophil adherence to cytokine-activated endothelial cell (EC) monolayers depends on the expression of the endothelial leukocyte adhesion molecule-1 (ELAM-1). The ligand for ELAM-1 is the sialylated Lewis-x antigen (SLe(x)) structure. The selectin LAM-1 (or LECAM-1) has been described as one of the SLe(x)-presenting glycoproteins involved in neutrophil binding to ELAM-1. Other presenter molecules have not yet been described. Our data demonstrate that the carcinoembryonic antigen (CEA)-like surface molecules on neutrophils--known as the nonspecific cross-reacting antigens (NCAs)--are involved in neutrophil adherence to monolayers of IL-1-beta-activated EC. The NCAs are recognized by CD66 (NCA-160 and NCA-90) and CD67 (NCA-95). Because NCA-95 and NCA-90 have previously been found to be phosphatidylinositol (PI)-linked, paroxysmal nocturnal hemoglobinuria (PNH) neutrophils (which lack PI-linked surface proteins) were tested as well. PNH neutrophils showed a diminished binding to activated EC. CD66 (on PNH cells still recognizing the transmembrane NCA-160 form) still inhibited the adherence of PNH cells to IL-1-beta-activated EC, but to a limited extent. Soluble CEA(-related) antigens inhibited normal neutrophil adherence as well, whereas neutrophil transmigration was unaffected. Sialidase-treatment as well as CD66 preclearing abolished the inhibitory capacity of the CEA(-related) antigens. The binding of soluble CEA antigens to IL-1-beta-pretreated EC was blocked by anti-ELAM-1. These soluble antigens, as well as the neutrophil NCA-160 and NCA-90, both recognized by CD66 antibodies, presented the SLe(x) determinant. Together, these findings indicate that the CD66 antigens (i.e., NCA-160/NCA-90) function as presenter molecules of the SLe(x) oligosaccharide structures on neutrophils that bind to ELAM-1 on EC.

Recombinant human interferon-gamma treatment in severe leucocyte adhesion deficiency.

We describe a patient with leucocyte adhesion deficiency (LAD). Clinically, the patient had delayed umbilical cord detachment, omphalitis, impaired wound healing and persistent leucocytosis. The patient had the severe form of LAD, with a total absence of leucocyte cell adhesion molecules (LeuCAMs) and undetectable mRNA for the beta chain, the common subunit of the LeuCAMs. In vitro neutrophil chemotaxis, aggregation and oxygen consumption were severely impaired. In vitro incubation of neutrophils with recombinant human interferon-gamma (rIFN-gamma) showed an increase in oxygen consumption, but no effect on the expression of the LeuCAMs, or the beta chain mRNA. In vivo treatment with IFN-gamma was started. The Fc gamma RI receptor appeared on the neutrophils, the LeuCAMs remained undetectable, while the neutrophil functions remained disturbed. The patient died of surgical complications after 10 weeks of rIFN-gamma treatment. No new infections or side-effects due to rIFN-gamma were observed.

Neutrophil migration across monolayers of resting or cytokine-activated endothelial cells. Role of intracellular calcium changes and fusion of specific granules with the plasma membrane.

Chemoattractants, used at concentrations to invoke optimal neutrophil chemotaxis, induce rapid changes in neutrophils such as a transient increase in intracellular Ca2+ ([Ca2+]i). We have previously observed that neutrophils adhering to cytokine-activated endothelial cells (EC) also respond with a rapid rise in [Ca2+]i caused by an endothelial membrane-bound form of platelet-activating factor. After preloading with the intracellular Ca(2+)-chelator bis-(O-aminophenoxyl)ethane-N,N,N',N'-tetraacetic acid (BAPTA/AM), neutrophils were no longer able to respond with a rapid rise in [Ca2+]i toward the chemoattractant FMLP or to rIL-1 beta-pretreated EC. These neutrophils were still able to adhere and migrate under the conditions tested. The only difference was that the BAPTA/AM-treated neutrophils migrated a little slower than untreated control neutrophils. This discrepancy was not observed at later time points. The BAPTA/AM-preloaded neutrophils did not differ from unloaded neutrophils in actin polymerization responses. Whereas untreated neutrophils demonstrated an up-regulation of the specific granule markers CD11b, CD45, and CD67 during migration (without any release from the azurophil granules), the BAPTA/AM pretreatment completely prevented this process. The BAPTA/AM-preloaded neutrophils did not release vitamin B12-binding protein from the specific granules upon treatment with FMLP. The down-modulation of the selectin member LAM-1, as seen upon neutrophil activation, was not affected by BAPTA/AM pretreatment of the neutrophils. Thus, neither the rapid rise in [Ca2+]i nor specific granule fusion with the plasma membrane constitute a prerequisite for neutrophil migration across resting or cytokine-activated EC.

Role of endothelial leukocyte adhesion molecule-1 and platelet-activating factor in neutrophil adherence to IL-1-prestimulated endothelial cells. Endothelial leukocyte adhesion molecule-1-mediated CD18 activation.

Adherence of neutrophils to endothelium is a key event in the sequence of inflammatory leukocyte responses. Double-color FACS analysis was used to determine the extent and kinetics of neutrophil adherence to rIL-1 beta-pretreated endothelial cells (EC). Neutrophils bound very avidly when the EC were prestimulated for 4 to 6 h with rIL-1 beta. Anti-ELAM-1 F(ab)2 fragments inhibited this adherence for more than 80%. On the other hand, anti-CD18 F(ab)2 fragments also inhibited the neutrophil adherence (40 to 50%). Combined use of anti-ELAM-1 and anti-CD18 F(ab)2 fragments completely prevented adherence. Neutrophils became activated as soon as they made contact with the rIL-1 beta-pretreated EC. First, neutrophils depleted of intracellular ATP showed a clearly decreased adherence completely dependent on ELAM-1-mediated binding, i.e., without additional effects of CD18 adhesion proteins. Thus, CD18 is activated during neutrophil adherence and then participates in the binding process. Secondly, the neutrophils responded with a transient rise in [Ca2+]i upon binding to rIL-1 beta-pretreated EC, which was demonstrated to be caused by endothelial cell-associated platelet-activating factor (PAF). However, the extent of neutrophil adherence to rIL-1 beta-pretreated EC was not affected by the use of the PAF-receptor antagonist WEB 2086, or removal of the EC-bound PAF. The only effect was a complete dependency of the neutrophil adherence on ELAM-1-mediated binding, although anti-CD18 mAb still induced 40 to 50% inhibition under these conditions. We therefore conclude that ELAM-1-mediated binding is the major mechanism for CD18 activation during neutrophil adherence to rIL-1 beta-pretreated EC.

Complement fragments C3b and iC3b coupled to latex induce a respiratory burst in human neutrophils.

The complement fragments C3b and iC3b were purified from human serum by affinity chromatography with Sepharose-coupled monoclonal antibody against the C3d region of C3. The resulting preparations were more than 95% pure and contained less than 0.1% native IgG. Purified C3b and iC3b were coupled to latex beads (0.8 micron diameter) by means of F(ab')2 fragments of monoclonal antibodies against the beta chain or the C3d region of C3, thus orienting the C3b and the iC3b on the latex with the C3b- and iC3b-specific regions outwards. These particles were found to activate the respiratory burst of freshly isolated human neutrophils to 20-30% of the maximal capacity. Latex particles randomly coated with C3b or iC3b were about 3 times less stimulatory. C3b, iC3b and IgG coupled to latex in an oriented fashion were about equally effective in stimulating the respiratory burst. Neutrophils from a patient with a total deficiency of CR3 responded normally to C3b-coated latex but did not respond to iC3b-coated latex. A monoclonal antibody against the alpha chain of CR3 inhibited the activation by iC3b-coated latex and a polyclonal antibody against CR1 partially inhibited the activation by C3b-coated latex. We found an additive effect between IgG-coated latex and C3b-coated latex, regardless of the presence of IgG and C3b on the same particle or on different particles. Thus, binding of ligands to either CR1 or CR3 per se is sufficient to induce an activating signal to the NADPH oxidase in human neutrophils.

Effects of sounds presented during general anaesthesia on postoperative course.

In a double-blind, randomized study, patients undergoing cholecystectomy were administered one of four different sounds during general anaesthesia: positive suggestions, nonsense suggestions, seaside sounds or sounds from the operating theatre. The effect of these sounds on the postoperative course was examined to assess intraoperative auditory registration. No differences were found between the four groups in postoperative variables.

Diagnostic evaluation and survival analysis of colorectal cancer patients with liver metastases.

Clinical features and their prognostic value were evaluated in 83 colorectal cancer patients with liver metastasis. The clinical features analysed included presenting symptoms and signs, liver function tests, extent of liver involvement, associated extrahepatic tumor growth, and physical condition of the patients. Overall median survival time after diagnosis of liver metastases was 8.4 months. Prognostic factors related to survival were symptoms, when referable to liver metastasis, and 5' Nt. Information is supplied to survey what selection of patients should be considered for various treatment options. A predominance of the patients showed bilobar liver involvement (79.6%), extrahepatic tumor growth (49.4%), or had an unresectable primary tumor (30.1%), thus leaving only 6% of the patients with liver metastases for surgical treatment with the intention of cure.

Scintimetric detection of choroidal malignant melanoma with [123I]-5-iodo-2-thiouracil.

[123I]-5-iodo-2-thiouracil (123I-ITU) was evaluated as a radiopharmaceutical for tumor detection in 10 patients with proven choroidal melanoma. Uptake of 123I-ITU was measured with a specially designed single eye probe collimator, 24 h after administration of 123I-ITU. Increased uptake in the tumor-bearing eye as compared to the fellow nontumor bearing eye was found in 7 out of 10 cases when the probe was located 3.5 cm in front of the eye (p less than 0.01). By using a double pinhole collimator tests were positive in 3 out of 10 123I-ITU studies only. Tests with 123I-ITU were compared with 67Ga tests in the same patients. The 67Ga tests with the single eye probe collimator were positive in 6 out 10 cases when the probe was located 6 cm in front of the eye. With the double pinhole collimator tests were positive in 7 out of 10 67Ga studies. It is concluded that 123I-labeled thiouracil is at least as useful as a radiopharmaceutical for ocular melanoma diagnosis as 67Ga-citrate, provided measurements are performed with a single eye probe.