Cloning of the IgM heavy chain of the bottlenose dolphin (Tursiops truncatus), and initial analysis of VH gene usage

Clones encoding the dolphin IgM heavy (micro) chain gene were isolated from a cDNA library of peripheral blood leukocytes. Genomic Southern blot analyses showed that the dolphin IGHM gene is most likely present in a single copy, and its sequence shows greatest similarity to those of the IGHM gene of the sheep, pig and cow, evolutionarily related artiodactyls. The transmembrane (TM) form of the IGHM chain was isolated by 3' RACE. While showing similarities to the TM regions of other mammalian IGHM chains, the highly conserved Ser residue of the CART motif is substituted with a Gly in the dolphin. In contrast to the pig and cow, which utilize only a single VH family, the dolphin expresses at least two distinct VH families, belonging to the mammalian VH clans I and III. At least two JH genes were identified in the dolphin. Some CDR3 regions of the dolphin VH are long (up to 21 amino acids), and contain multiple Cys residues, hypothesized to stabilize the CDR3 structure through disulfide bond formation.     

Subepithelial neuroendocrine cells and carcinoid tumours of the human small intestine and appendix. A comparative immunohistochemical study with regard to serotonin, neuron-specific enolase and S-100 protein reactivity

A comparative immunocytochemical study was performed of subepithelial neuroendocrine cells of the human small intestine and appendix and carcinoid tumours of these sites, using a monoclonal antibody to serotonin and polyclonal antisera against neuron-specific enolase (NSE) and S-100 protein. Subepithelial neuroendocrine cells were easily identified in the lamina propria of the appendix. These cells, which sometimes occurred in aggregates, displayed serotonin and NSE immunoreactivity and were surrounded by S-100 protein immunoreactive cells, presumably of Schwann cell origin. In the appendix scattered cells with corresponding morphological features and immunoreactivity were also observed deep in the submucosa. In addition, subepithelial neuroendocrine cells were sparsely present in the lamina propria of the small intestine, occurring only as single cells in the deeper part of the mucosa below or between the epithelial crypts. Most appendiceal carcinoid tumours (11 of 12 examined cases) were biphasic and consisted of neuroendocrine tumour cells with intermingled S-100 protein immunoreactive cells (Schwann cells) with long cytoplasmic extensions. However, small intestinal (11 cases) and caecal (10 cases) carcinoids lacked S-100 protein immunoreactive cells as an integral component. The results indicate that the appendiceal carcinoids are mostly closely related structurally to the subepithelial neuroendocrine and Schwann cell aggregates of the lamina propria and are thus presumed to be histogenetically related to this cell system, while the histogenesis of small-intestinal and caecal carcinoids remains less clear.     

Effects of human plasma proteins on maturation of monocyte-derived dendritic cells

Dendritic cells (DC) are a promising tool for vaccine therapy due to their unique properties as antigen presenting cells and their ability to prime naïve T cells. Increasing evidence suggests that maturation stage of DC critically influences the fate of the immune response. Generation of monocyte-derived DC for clinically applicable immunotherapy requires the use of well-defined components and stringent culture conditions. An alternative strategy is to use human autologous serum. However, its constituents are not stable and reflect the inflammatory condition of the donor. In order to investigate whether DC properties are influenced by proteins present in the plasma, we matured human monocyte-derived DC with four main plasma components: fibrinogen, fibronectin, plasminogen or C-reactive protein. These purified proteins were added at various concentrations on day 6 after the initial differentiation induced by IL-4 and GM-CSF. The maturation was assessed by phenotyping of maturation-associated marker (CD83) and co-stimulatory molecule CD86 as well as IL-12 production. Functional properties of DC were assessed by endocytic activity and mixed leukocyte culture. Our results indicate that fibrinogen had DC-maturation effect comparable to poly-I:C, TNF-α and PGE₂ as a positive control, but it failed to induce IL-12 production. The other plasma proteins had no effect on DC maturation. CRP at high concentration had rather inhibitory effect on DC induced lymphocyte function. We conclude that none of the tested plasma components and acute phase proteins sufficiently induce fully competent mature DC. This finding is important for the preparation of human DC-based vaccines supplemented by autologous sera.     

Brief report: the Brazelton Neonatal Behavioral Assessment Scale detects differences among newborn infants of optimal health

OBJECTIVE: To determine whether the Brazelton Neonatal Behavioral Assessment Scale (NBAS) can detect behavioral differences in newborn infants of optimal health and, if such differences appear, also detect gender differences among those neonates. METHODS: Participants were a group of healthy Swedish neonates, 20 boys and 18 girls. The infants were assessed by the NBAS under standardized conditions at 48-72 hours of age, at the midpoint between two meals. RESULTS: All items except those in the dimensions Autonomic System and Motor System had a wide interquartile range. The trend was that girls had higher median item profiles, which means a higher level of functioning than boys. Four out of seven median values in the dimension Social Interactive Organization, as well as the median value in the self-quieting item in the dimension State Regulation, were significantly higher for girls. The interquartile range of the items seemed wider for boys than for girls. CONCLUSIONS: The results indicate behavioral variability among healthy neonates. Gender differences were also observed with girls showing higher levels of functioning than boys.     

Anxiety and Depression Symptoms Among Farmers: The HUNT Study,Norway

Agriculture has undergone profound changes, and farmers face a wide variety of stressors. Our aim was to study the levels of anxiety and depression symptoms among Norwegian farmers compared with other occupational groups. Working participants in the HUNT3 Survey (The Nord-Trøndelag Health Study, 2006–2008), aged 19–66.9 years, were included in this cross-sectional study. We compared farmers (women, n = 317; men, n = 1,100) with HUNT3 participants working in other occupational groups (women, n = 13,429; men, n = 10,026), classified according to socioeconomic status. We used the Hospital Anxiety and Depression Scale (HADS) to measure anxiety and depression symptoms. Both male and female farmers had higher levels of depression symptoms than the general working population, but the levels of anxiety symptoms did not differ. The differences in depression symptom levels between farmers and the general working population increased with age. In an age-adjusted logistic regression analysis, the odds ratio (OR) for depression caseness (HADS-D ≥8) when compared with the general working population was 1.49 (95% confidence interval [CI]: 1.22–1.83) in men and 1.29 (95% CI: 0.85–1.95) in women. Male farmers had a higher OR of depression caseness than any other occupational group (OR = 1.94, 95% CI: 1.52–2.49, using higher-grade professionals as reference). Female farmers had an OR similar to men (2.00, 95% CI: 1.26–3.17), but lower than other manual occupations. We found that farmers had high levels of depression symptoms and average levels of anxiety symptoms compared with other occupational groups.     

Quality of community-based day centre services for people with psychiatric disabilities: psychometric properties of the Quality in Psychiatric Care – Daily Activities (QPC–DA)

Background/aims The aim of the present study was to test the psychometric properties and dimensionality of the instrument Quality in Psychiatric Care – Daily Activities (QPC–DA) and to briefly describe the day centre attendees’ perception of the quality at community-based day centre services. Methods A sample of 218 attendees from 14 community-based day centre services in seven municipalities in Sweden participated in the study. Results Confirmatory factor analysis revealed that the QPC–DA consists of six dimensions and has a factor structure that to a large extent corresponds to that found in other studies of quality in psychiatric care settings, such as inpatient, outpatient, forensic inpatient, and housing support for people with psychiatric disabilities. The internal consistency of the factors was satisfactory and thus the QPC–DA showed adequate psychometric properties. The attendees’ ratings of quality of community-based day centre services were generally high. The highest rating was for the encounter dimension and the lowest for the participation and the secluded environment dimensions, indicating areas for improvement. Conclusion/significance The QPC–DA includes important aspects of the attendees’ assessment of quality of community-based day centre services and offers a simple and inexpensive way to evaluate quality from their perspective.     

The clinical value of serum S-100 protein measurements in minor head injury: a Scandinavian multicentre study

PURPOSE: This study of patients with minor head injury was designed to investigate the relation of S-100 protein measurements to computed tomograpy (CT) findings and patients outcomes. Increased serum levels of this protein were hypothetized to predict intracranial pathology and increased frequency of post-concussion symptoms. METHODS: One hundred and eighty-two patients were studied with Glasgow Coma Scale scores of 13-15. The study recruited patients from three Scandinavian neurotrauma centres. Serum levels of S-100 protein were measured at admittance and CT scans of the brain were obtained within 24 hours postinjury in all patients. Outcome was evaluated with the Rivermead Postconcussion Symptoms Questionnaire (RPQ) 3 months after the injury. RESULTS: Increased serum level of S-100 protein was detected in 69 (38%) patients, and CT scan demonstrated intracranial pathology in 10 (5%) (brain contusion in seven, epidural haematoma in two, traumatic subarachnoid haemorrhage in one). The proportion of patients with detectable serum level was significantly (p < 0.01) higher among those with intracranial pathology (90%) compared to those without (35%). The negative predictive value of an undetectable S-100 level was 0.99. Sixty-two per cent reported one or more post-concussion symptoms at follow-up. A trend was observed towards an increased frequency of post-concussion symptoms among patients with detectable serum levels. CONCLUSIONS: Undetectable serum level of S-100 protein predicts normal intracranial findings on CT scan. Determination of S-100 protein in serum may be used to select patients for CT scanning. Increased S-100 serum levels may be more related to post-concussion symptoms caused by mild traumatic brain injury than to symptoms of psychological origin.     

Cellular processing of (125)I- and (111)in-labeled epidermal growth factor (EGF) bound to cultured A431 tumor cells

Low molecular weight of epidermal growth factor (EGF) enables better intratumoral penetration in comparison with larger targeting proteins, but the cellular retention of EGF-associated radioactivity is poor for directly iodinated EGF. An attempt was made to improve intracellular retention by the use of metal-diethylenetriaminepentaacetic acid or nonphenolic linker (N-succinimidyl-para-iodobenzoate) as labeling agents. The use of nonphenolic linker did not improve retention of the radioactivity in A431 carcinoma cell line. The use of the radiometal label provided an appreciable prolongation of radioactivity residence inside the cell.     

Kinetics of 76Br-labeled anti-CEA antibodies in pigs; aspects of dosimetry and PET imaging properties

A monoclonal antibody labeled with the positron-emitting radionuclide 76Br (T(1/2) 16.2 h) has previously been shown useful for positron emission tomography (PET) imaging of experimental tumors. Our aim in the present study was to investigate the effects of the complex decay scheme of this radionuclide on normal organ dosimetry and PET image quality. Three mini-pigs were injected intravenously with 46-75 MBq of the 76Br-labeled anti-CEA antibody 38S1, and the whole-body kinetics followed by PET imaging for 19 h. From PET data, absorbed doses in human organs were estimated using the MIRDOSE 3.0 software. The highest 76Br concentrations were found in lungs, after a correction for the air volume in this organ. The lungs received the highest absorbed dose (mGy/MBq, mean+/-maximum error), 0.84+/-0.16, followed by liver, 0.74+/-0.28, and small intestine, 0.55+/-0.05, while the effective dose equivalent was 0.41+/-0.03 mSv/MBq. The PET imaging properties of 76Br in a two-dimensional 2D PET camera, including central area resolution and scattering effects, were investigated in phantoms and compared to those of 18F. In a 0.97 g/cm3 material, approximating soft tissue density, the FMHW ("full width at half-maximum") value of the point spread function was 7.7+/-0.2 mm for 76Br and 6.0+/-0.1 mm for 18F. In conclusion, radioimmuno PET using 76Br-labeled antibodies resulted in a fairly even distribution of the radiation dose, where the highest absorbed organ doses were only about two to three times higher than the mean absorbed body dose. The high energy beta+ spectrum in the 76Br decay had only minor effects on the resolution, but may decrease the quantification accuracy, especially in organs with a lower density such as a lung.     

Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts

Nanoparticles in a biological fluid (plasma, or otherwise) associate with a range of biopolymers, especially proteins, organized into the "protein corona" that is associated with the nanoparticle and continuously exchanging with the proteins in the environment. Methodologies to determine the corona and to understand its dependence on nanomaterial properties are likely to become important in bionanoscience. Here, we study the long-lived ("hard") protein corona formed from human plasma for a range of nanoparticles that differ in surface properties and size. Six different polystyrene nanoparticles were studied: three different surface chemistries (plain PS, carboxyl-modified, and amine-modified) and two sizes of each (50 and 100 nm), enabling us to perform systematic studies of the effect of surface properties and size on the detailed protein coronas. Proteins in the corona that are conserved and unique across the nanoparticle types were identified and classified according to the protein functional properties. Remarkably, both size and surface properties were found to play a very significant role in determining the nanoparticle coronas on the different particles of identical materials. We comment on the future need for scientific understanding, characterization, and possibly some additional emphasis on standards for the surfaces of nanoparticles.