Nanoscale patterning of self-assembled monolayer (SAM)-functionalised substrates with single molecule contact printing.

Defined arrangements of individual molecules are covalenty connected ("printed") onto SAM-functionalised gold substrates with nanometer resolution. Substrates were initially pre-functionlised by coating with 3,3'-dithiodipropionic acid (DTPA) to form a self-assembled monolayer (SAM), which was characterised by atomic force microscopy (AFM), contact angle goniometry, cyclic voltammetry and surface plasmon resonance (SPR) spectroscopy. Pre-defined "ink" patterns displayed on DNA origami-based single-use carriers ("stamp") were covalently conjugated to the SAM using 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) and N-hydroxy-succinimide (NHS). These anchor points were used to create nanometer-precise single-molecule arrays, here with complementary DNA and streptavidin. Sequential steps of the printing process were evaluated by AFM and SPR spectroscopy. It was shown that 30% of the detected arrangements closely match the expected length distribution of designed patterns, whereas another 40% exhibit error within the range of only 1 streptavidin molecule. SPR results indicate that imposing a defined separation between molecular anchor points within the pattern through this printing process enhances the efficiency for association of specific binding partners for systems with high sterical hindrance. This study expands upon earlier findings where geometrical information was conserved by the application of DNA nanostructures, by establishing a generalisable strategy which is universally applicable to nearly any type of prefunctionalised substrate such as metals, plastics, silicates, ITO or 2D materials.

New experimental perspectives for soft x-ray absorption spectroscopies at ultra-low temperatures below 50 mK and in high magnetic fields up to 7 T.

A new ultra-low temperature experiment including a superconducting vector magnet has been developed for soft x-ray absorption spectroscopy experiments at third generation synchrotron light sources. The sample is cooled below 50 mK by a cryogen free (3)He-(4)He dilution refrigerator. At the same time, magnetic fields of up to ±7 T in the horizontal direction and ±0.5 T in the vertical direction can be applied by a superconducting vector magnet. The setup allows to study ex situ and in situ prepared samples, offered by an attached UHV preparation chamber with load lock. The transfer of the prepared samples between the preparation section and the dilution refrigerator is carried out under cryogenic temperatures. First commissioning studies have been carried out at the Variable Polarization XUV Beamline P04 at PETRA III and the influence of the incident photon beam to the sample temperature has been studied.

Energy and Protein Intake, Anthropometrics, and Disease Burden in Elderly Home-care Receivers--A Cross-sectional Study in Germany (ErnSIPP Study).

OBJECTIVE: To date, no study has examined the nutritional status and disease burden of elderly home-care receivers living in Germany. Aim of this cross-sectional study was, first, to assess disease burden and nutritional status, denoted in anthropometrics, and, second, to investigate associations between anthropometrics and disease burden. DESIGN: Cross-sectional multi-centre study. SETTING: Home-care receivers living in three urban areas of Germany in 2010. PARTICIPANTS: 353 elderly (>64 years) in home care (128 males aged 79.1 ±7.8 years, 225 females aged 82.0 ±7.5 years). MEASUREMENTS: Nutritional status was assessed by body mass index (BMI), mid upper arm circumference (MUAC) and calf circumference (CC). Medical conditions were assessed in personal interviews. A 3-day prospective nutrition diary was kept. Metric data are reported as mean±SD or median (interquartile range), p<0.05 was considered significant. RESULTS: Most participants were substantially (59%), and 11% severest in need of care. The seniors suffered from 5 (4-7) chronic diseases; dementia, depression, stroke, and respiratory illness were most prevalent (each 20-40%). More than one-third of participants had only moderate or poor appetite, nearly half were unable to eat independently. Chewing problems were reported for 52% of study participants, and more than one quarter of elderly had swallowing problems. Daily mean energy intake was 2017±528 kcal in men (n=123) and 1731±451 kcal in women (n=216; p<0.001). Mean protein intake amounted to 1.0 g/kg body weight. Mean BMI was 28.2±6.2 kg/m² (n=341), 14% of seniors had a BMI <22 kg/m² (including 4% with BMI <20 kg/m²). Critical MUAC (<22 cm) was indicated in 6% of subjects; and CC <31 cm in 11% of men, 21% of women (p<0.05). After adjusting for sex and age, BMI, MUAC and CC were negatively associated with high care level, hospitalization in the previous year, nausea/vomiting, prevalence of dementia, poor appetite, and eating difficulties like dependency, chewing and swallowing problems. CONCLUSION: We recommend to pay special attention to the nutritional status of elderly persons in home-care exhibiting named disease burden.

Stability and cell uptake of calcium carbonate templated insulin microparticles.

Therapeutic proteins are an integral part of today's pharmaceutical practice, but they still present challenges from the drug delivery point of view. In this work, a new approach is studied based on hard templating for fabrication of microparticles composed of pure insulin, which may enable effective delivery, for instance pulmonary delivery. The approach is both simple and versatile: the protein particles are prepared by selective precipitation into porous CaCO3 microtemplates, followed by full decomposition of the template at the isoelectric point of the protein (pH 5.2). Control over the main material parameters (mechanical properties, porosity, morphology and stability at physiological conditions) are critical for the envisioned application in drug delivery. It is demonstrated that these critical parameters can be significantly tuned by a slight final pH variation around the isoelectric point (pH range 4-6) and by the denaturation degree of insulin. Electrostatic interactions and inter-protein crosslinking in the protein particles as well as their internal structure are considered, to explain the variation in the particle properties. The particle property parameters are explored using atomic force microscopy, optical microscopy and circular dichroism spectra. Finally, phagocytic clearance of the protein particles in vitro was studied to explore possible enhancements in particle fabrication to improve the efficiency of insulin delivery by inhalation.

Malnutrition is related to functional impairment in older adults receiving home care.

OBJECTIVES: The aims of this work were (a) to provide a detailed description of the association between nutritional (Mini Nutritional Assessment; MNA®) and functional status in a sample of older adults receiving home care, using both questionnaire- and performance-based functional methods, and (b) to investigate the impact of different MNA subscales on this association. DESIGN: Multi-centre, cross-sectional. SETTING: Home care. PARTICIPANTS: 296 persons ≥65 years in need of care (80.7±7.7 y). MEASUREMENTS: Nutritional status was determined by the MNA and functional status by two questionnaires (Instrumental and Basic Activities of Daily Living; IADL, ADL) and three performance tests (handgrip strength, HGS; Short Physical Performance Battery, SPPB; Timed 'Up and Go' Test, TUG). A categorical and a covariance analytical approach were used to test for differences in functional status between MNA groups (well nourished, risk of malnutrition, malnourished). In addition, functional parameters were correlated with total MNA, a modified MNA version (modMNA), where functional items were excluded, and MNA subscales ('functionality', 'general assessment', 'anthropometry', 'dietary assessment', and 'subjective assessment'). RESULTS: 57% of the participants were at risk of malnutrition and 12% malnourished. 35% reported severe limitations in IADL, 18% in ADL. 40%, 39% and 35% had severe limitations in HGS, SPPB and TUG; 9%, 28% and 34% were not able to perform the tests. Functional status deteriorated significantly from the well nourished to the malnourished group in all functional measures. The modMNA was weak but still significantly related to all functional parameters except TUG. The subscale 'functionality' revealed strongest correlations with functional measures. All other MNA subscales showed only weak or no associations. CONCLUSION: More than one half of the seniors receiving home care were at nutritional risk and poor functional level, respectively. Malnutrition according to MNA was significantly associated to both questionnaire- and performance-based functional measures even after exclusion of functional MNA items.

Grading evidence and recommendations for clinical practice guidelines in nephrology. A position statement from Kidney Disease: Improving Global Outcomes (KDIGO).

Considerable variation in grading systems used to rate the strength of guideline recommendations and the quality of the supporting evidence in Nephrology highlights the need for a uniform, internationally accepted, rigorous system. In 2004, Kidney Disease: Improving Global Outcomes (KDIGO) commissioned a methods expert group to recommend an approach for grading in future nephrology guidelines. This position statement by KDIGO recommends adopting the Grades of Recommendation Assessment, Development, and Evaluation (GRADE) approach for the grading of evidence and guidelines on interventions. The GRADE approach appraises systematic reviews of the benefits and harms of an intervention to determine its net health benefit. The system considers the design, quality, and quantity of studies as well as the consistency and directness of findings when grading the quality of evidence. The strength of the recommendation builds on the quality of the evidence and additional considerations including costs. Adaptations of the GRADE approach are presented to address some issues pertinent to the field of nephrology, including (1) the need to extrapolate from studies performed predominantly in patients without kidney disease, and (2) the need to use qualitative summaries of effects when it is not feasible to quantitatively summarize them. Further refinement of the system will be required for grading of evidence on questions other than those related to intervention effects, such as diagnostic accuracy and prognosis.

New approaches to the treatment of calcium and phosphorus abnormalities in patients on hemodialysis.

Disturbances in calcium and phosphorus are common in patients who are on chronic hemodialysis. Current therapy consists mainly of administration of calcium-containing phosphate binders and calcitriol. However, the use of these therapies has been limited by both iatrogenic hypercalcemia and hyperphosphatemia. Indeed, hyperphosphatemia and elevations in the calcium x phosphorus product have recently been linked to increased mortality and a high burden of cardiac calcifications on electron-beam computed tomography. Novel therapeutic agents such as sevelamer and vitamin D analogs have thus been developed with the goal of decreasing oral calcium loading and hypercalcemia. We provide an algorithm for the management of calcium and phosphorus abnormalities, based on newer recommendations for tighter calcium and phosphorus control with a lower target for the calcium x phosphorus product.

AL-amyloidosis of the kidney initially presenting as minimal change glomerulonephritis.

Small amounts of amyloid in kidney biopsy specimens may be missed on routine examination unless specifically targeted. Occasionally, this oversight results in a diagnosis of minimal change glomerulonephritis (MCGN). This misdiagnosis may be facilitated by the fact that typical "minimal changes" with flattening and effacement of the epithelial foot processes can be found in capillary loops directly affected by amyloid deposition as well as in capillary loops of glomeruli with only mild amyloid deposition in the mesangium. Repeatedly, the diagnosis of MCGN had to be corrected to renal amyloidosis when re-examination by special techniques succeeded in detecting even small amounts of amyloid fibrils. We present the case of a previously healthy 49-year-old man who suddenly developed nephrotic syndrome. A first renal biopsy showed MCGN. Proteinuria remained refractory to immunosuppressive treatments, and creatinine clearance deteriorated rapidly. Two years later, a repeat renal biopsy showed AL-amyloidosis. In this case, re-examination of the first biopsy in the light of the final diagnosis again did not show any deposition of amyloid fibrils. We suspect that proteinuria and epithelial podocyte changes in amyloidosis are caused by factors other than deposition of amyloid fibrils itself. Possibly a cytokine release during the early fibril formation leads to abnormalities even before the typical structural changes of renal amyloidosis can be detected. This is analogous to the hypothesis of a circulating factor that leads to proteinuria in focal segmental glomerulosclerosis or the speculation of altered lymphokine expression associated with the development of MCGN in Hodgkin's disease.

Predominance of mononuclear cells expressing the chemokine receptor CCR5 in synovial effusions of patients with different forms of arthritis.

OBJECTIVE: To study the role of the chemokine receptors CCR5 and CCR2 in patients with arthritis. METHODS: CCR5 expression on peripheral blood leukocytes was compared with the expression on leukocytes isolated from the synovial fluid of 20 patients with different rheumatic joint diseases. Three additional samples were studied for CCR2 expression. The expression of chemokine receptors on blood and synovial fluid leukocytes was determined by 3-color flow cytometry analysis. To test CCR5 receptor down-modulation from the cell surface, leukocytes were incubated in vitro with a RANTES (regulated on activation, normal T cell expressed and secreted) derivative, aminooxypentane (AOP)-RANTES. Patients were genotyped for the delta32 CCR5 deletion by polymerase chain reaction. RESULTS: A high percentage of CCR5-expressing CD4+ and CD8+ T cells (74% and 81%, respectively), monocytes (51%), and natural killer cells (35%) was found in the synovial fluid of all patients, whereas in the peripheral blood, only a small percentage of these cells expressed CCR5 (13%, 32%, 7.8%, and 4%, respectively). Infiltration of CCR5-positive leukocytes was not reduced in CCR5-heterozygous patients. A similar, but less pronounced, distribution was observed for CCR2-positive T cells. In vitro, CCR5 was completely down-modulated on synovial fluid leukocytes by AOP-RANTES. CONCLUSION: The predominance of CCR5-positive mononuclear cells in the synovial effusions of patients with arthritis suggests an important role for CCR5 in the process of joint inflammation, and identifies CCR5 as a possible new target for therapeutic intervention.

Prevention of corticosteroid-induced suppression of human polymorphonuclear leukocyte-induced damage of Aspergillus fumigatus hyphae by granulocyte colony-stimulating factor and gamma interferon.

Neutrophils (PMNs) are a critical line of defense against Aspergillus fumigatus infection. Increased frequency of invasive aspergillosis has been observed in patients receiving corticosteroids, suggesting a deleterious effect of these compounds on PMN antifungal function. To investigate this hypothesis and to determine the potential preventive utility of granulocyte colony-stimulating factor (G-CSF) and gamma interferon (IFN-gamma), the effects of hydrocortisone (HCS) and dexamethasone (DXS) on PMN-induced damage of Aspergillus fumigatus hyphae were studied with or without pretreatment of PMNs with G-CSF and IFN-gamma. PMNs treated with HCS (> or = 3,000 microM) or DXS (> or = 10 microM) during a 2-h colorimetric tetrazolium metabolic assay (using methylthiotetrazolium) showed suppressed percentage of hyphal damage (P < 0.02). In addition, both HCS (> or = 30 microM) and DXS (> or = 1 microM) significantly suppressed oxidative burst measured as superoxide anion release by PMNs in response to opsonized and nonopsonized hyphae as well as to N-formylmethionyl leucyl phenylalanine. Pretreatment of PMNs with G-CSF (4,000 U/ml) and/or IFN-gamma (100 and 1,000 U/ml) for 90 min prevented the suppression of hyphal damage that occurred in the presence of HCS (3,000 microM; P < 0.01) or DXS (10 microM; P < or = 0.001). G-CSF (4,000 U/ml) and IFN-gamma (100 U/ml) combined had an additive effect on increasing the antifungal activity of HCS-treated but not of DXS-treated PMNs compared with IFN-gamma alone (P = 0.015). Thus, these findings reveal that corticosteroids impair PMN function in response to A. fumigatus and that G-CSF and IFN-gamma prevent this impairment.

Enhancement of oxidative response and damage caused by human neutrophils to Aspergillus fumigatus hyphae by granulocyte colony-stimulating factor and gamma interferon.

Invasive aspergillosis is a serious fungal infection caused by the proliferation and invasion of Aspergillus hyphae in tissue. Neutrophils (PMNs) are the most important line of defense against Aspergillus hyphae. To investigate the role of granulocyte colony-stimulating factor (G-CSF) and gamma interferon (IFN-gamma) against Aspergillus fumigatus, we studied the effects of the two cytokines on the oxidative burst and the capacity of normal human PMNs to damage hyphae of the organism. G-CSF enhanced PMN oxidative burst measured as superoxide anion (O2-) production in response to N-formylmethionyl leucyl phenylalanine, serum opsonized hyphae, and nonopsonized hyphae by 75, 37, and 24%, respectively, compared with control PMNs (P < 0.015). IFN-gamma also induced increases of 52, 71, and 96%, respectively, in response to the same stimuli (P < 0.006). In addition, the capacity of PMNs to damage hyphae as measured by the 3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MMT) colorimetric metabolic assay was significantly enhanced by G-CSF and IFN-gamma (P < 0.01 and < 0.05, respectively). The enhancement was achieved irrespective of serum opsonization of the hyphae, suggesting upregulatory actions of the two cytokines on signal pathways specific for opsonized and nonopsonized hyphae. The combination of the two cytokines exhibited an additive effect at the higher concentrations compared with the effects of the cytokines alone (P < 0.05). Pretreatment of PMNs with protein synthesis inhibitors showed that IFN-gamma activates PMN function through transcriptional regulation, whereas the effect of G-CSF does not require new proteins. These in vitro effects suggest modulatory roles for G-CSF and IFN-gamma in the host defense against Aspergillus hyphae irrespective of serum opsonization and a potential utility of the cytokines as adjuncts for the prevention and possible treatment of invasive aspergillosis.

Neutrophil oxidative burst in response to blastoconidia and pseudohyphae of Candida albicans: augmentation by granulocyte colony-stimulating factor and interferon-gamma.

The effects of granulocyte colony-stimulating factor (G-CSF) and interferon-gamma (IFN-gamma) on the oxidative burst of neutrophils (PMNL) in response to blastoconidia and pseudohyphae of Candida albicans were assessed and compared with those in response to N-FMLP. G-CSF enhanced oxidative burst, as measured by superoxide production, in response to both FMLP and opsonized blastoconidia. The enhancement of oxidative burst in response to FMLP was significantly greater (P = .004) than that in response to blastoconidia (65% and 39%, respectively). G-CSF also enhanced oxidative burst in response to pseudohyphae. IFN-gamma enhanced oxidative burst in response to FMLP and opsonized blastoconidia by 53% and 50%, respectively. Moreover, IFN-gamma significantly enhanced oxidative burst in response to opsonized and nonopsonized hyphae by 86% and 65%, respectively. These results demonstrate that G-CSF and IFN-gamma enhance the oxidative burst of PMNL in response to both blastoconidia and pseudohyphae of C. albicans and suggest an immunomodulatory role of the two cytokines in the host defenses against this fungus.

[Early detection of colorectal cancers by detection of occult blood in feces using Krypto-Haem SSW]. / Früherkennung kolorektaler Karzinome durch den Nachweis von okkultem Blut im Stuhl mit Krypto-Haem SSW.

Screening for cancer of the bowels with tests for occult blood in the stools obtained an increasing interest during the last 15 years. In the GDR Krypto-Haem SSW is at the disposal for this purpose. Up to now two larger investigations were performed with this preparation. The results of the studies with Krypto-Haem and Haemoccult were compared as well as questions of the test performance, the secondary diagnostics, the sensitivity of the tests, the compliance and organisation of a screening are treated. From this derived that Krypto-Haem SSW can be recommended as diagnostic test for colorectal tumours within the basic medical care.

[Colonic cancer screening with KRYPTO-HAEM SSW. The results in 3015 persons]. / Dickdarmkarzinom-Screening mit KRYPTO-HAEM SSW. Ergebnisse an 3015 Personen.

In a factory 3015 individuals have been tested for occult blood in stool using the KRYPTO-HAEM SSW. Test results were positive in 100 individuals (3.3%). Subsequent investigations have revealed 6 carcinomas of the colon, one carcinoid in the cecum and adenomas in 9 cases. Eleven out of the 16 tumors detected were beyond the range of the rectoscope. The test was valued as false-positive in 42 individuals (1.4%). A cecum carcinoma and 5 adenomas were revealed by false-negative findings. The importance of case history and exact diagnostic clarification of a positive test is pointed out. The inclusion of the KRYPTO-HAEM test into basic medical care proved to be an adequate measure for early detection of colorectal tumors.

[Screening for colorectal cancer with a test for occult blood in stool (author's transl)]. / Screening auf kolorektalen Krebs mit einem Suchtest auf okkultes Blut im Stuhl.

Results of an occult-blood-test in 1 072 persons are reported. The test, recently registered as KRYPTO-HAEM, was developed by the Central Institute of Molecular Biology of the Academy of Sciences of the GDR. 27 patients (2.5%) had positive findings and by clinical examination 1 carcinoma of the sigmoid colon and 2 adenomas were detected. In 1 case (adenoma of the rectum) the test had a false-negative result. The findings were compared with own experiences with the HAEMOCCULT-Test. The KRYPTO-HAEM-Test is recommended for screening for colorectal tumors.