A combination of low-dose bevacizumab and imatinib enhances vascular normalisation without inducing extracellular matrix deposition.

BACKGROUND: Vascular endothelial growth factor (VEGF)-targeting drugs normalise the tumour vasculature and improve access for chemotherapy. However, excessive VEGF inhibition fails to improve clinical outcome, and successive treatment cycles lead to incremental extracellular matrix (ECM) deposition, which limits perfusion and drug delivery. We show here, that low-dose VEGF inhibition augmented with PDGF-R inhibition leads to superior vascular normalisation without incremental ECM deposition thus maintaining access for therapy. METHODS: Collagen IV expression was analysed in response to VEGF inhibition in liver metastasis of colorectal cancer (CRC) patients, in syngeneic (Panc02) and xenograft tumours of human colorectal cancer cells (LS174T). The xenograft tumours were treated with low (0.5 mg kg-1 body weight) or high (5 mg kg-1 body weight) doses of the anti-VEGF antibody bevacizumab with or without the tyrosine kinase inhibitor imatinib. Changes in tumour growth, and vascular parameters, including microvessel density, pericyte coverage, leakiness, hypoxia, perfusion, fraction of vessels with an open lumen, and type IV collagen deposition were compared. RESULTS: ECM deposition was increased after standard VEGF inhibition in patients and tumour models. In contrast, treatment with low-dose bevacizumab and imatinib produced similar growth inhibition without inducing detrimental collagen IV deposition, leading to superior vascular normalisation, reduced leakiness, improved oxygenation, more open vessels that permit perfusion and access for therapy. CONCLUSIONS: Low-dose bevacizumab augmented by imatinib selects a mature, highly normalised and well perfused tumour vasculature without inducing incremental ECM deposition that normally limits the effectiveness of VEGF targeting drugs.

Correction: Role of growth arrest-specific gene 6-mer axis in multiple myeloma.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Mast cells decrease efficacy of anti-angiogenic therapy by secreting matrix-degrading granzyme B.

Resistance towards VEGF-centered anti-angiogenic therapy still represents a substantial clinical challenge. We report here that mast cells alter the proliferative and organizational state of endothelial cells which reduces the efficacy of anti-angiogenic therapy. Consequently, absence of mast cells sensitizes tumor vessels for anti-angiogenic therapy in different tumor models. Mechanistically, anti-angiogenic therapy only initially reduces tumor vessel proliferation, however, this treatment effect was abrogated over time as a result of mast cell-mediated restimulation of angiogenesis. We show that mast cells secrete increased amounts of granzyme b upon therapy, which mobilizes pro-angiogenic laminin- and vitronectin-bound FGF-1 and GM-CSF from the tumor matrix. In addition, mast cells also diminish efficacy of anti-angiogenic therapy by secretion of FGF-2. These pro-angiogenic factors act beside the targeted VEGFA-VEGFR2-axis and reinduce endothelial cell proliferation and angiogenesis despite the presence of anti-angiogenic therapy. Importantly, inhibition of mast cell degranulation with cromolyn is able to improve efficacy of anti-angiogenic therapy. Thus, concomitant mast cell-targeting might lead to improved efficacy of anti-angiogenic therapy.Resistance towards VEGF-centered anti-angiogenic therapy is an important clinical challenge. Here, the authors show that mast cells mediate resistance to anti-angiogenetic inhibitors by altering the proliferative and organizational state of endothelial cells through mobilization of FGF-1 and GM-CSF from the tumor matrix and secretion of FGF-2.

Gender, autoantibodies, and obesity in newly diagnosed diabetic patients aged 40-75 years.

OBJECTIVE: To evaluate the frequency of autoimmune markers (islet cell antibodies (ICA] and glutamic acid decarboxylase antibodies [GADA]) and clinical features in newly diagnosed people with diabetes aged 40-75 years. RESEARCH DESIGN AND METHODS: Two hundred fifty-nine consecutive patients (aged 40-75 years) with newly suspected diabetes diagnosed during a 2-year period were studied. The diagnosis of newly discovered diabetes was confirmed in 203 patients. Gender, BMI, HbA1c, fasting C-peptide, ICA, and GADA were evaluated. The frequency of obesity was estimated using two different sets of criteria: 1) National Diabetes Data Group (NDDG) criteria, and 2) criteria based on a Swedish reference population. RESULTS: The annual incidence of diabetes was 106 per 100,000 people. The incidence of diabetes in those patients who were 40-54 years old was significantly higher in men than in women (odds ratio: 2.16; P = 0.001). ICA were detected in 16 of 203 patients (8%), whereas 17 of 203 patients (8%) were GADA+; 10 of 203 (5%) patients were positive for both ICA and GADA. Among the 203 diabetic patients, 19 (9.4%) were classified as having IDDM, giving an IDDM incidence of 10 per 100,000 people aged 40-75 years. The frequency of obesity in NIDDM was high but varied with its definition; the frequency of obesity was highest (P < 0.001) when NDDG criteria, and not Swedish reference values, were used (57 of 75 [76%] vs. 40 of 75 [53%] for women and 66 of 109 [61%] vs. 45 of 109 [41%] for men). CONCLUSIONS: A striking male preponderance was found among incident cases of diabetes in people aged 40-54 years. Autoimmune markers were detected in 10% of incident cases of diabetes in people aged 40-75 years. Using a conservative estimation, as many as 10 of 100,000 middle-aged and elderly subjects developed IDDM. The frequency of obesity in NIDDM was high but this was also the case in the reference population.

Detection of autonomic sympathetic dysfunction in diabetic patients. A study using laser Doppler imaging.

OBJECTIVE: To study signs of the disturbed reflex autonomic sympathetic nerve function in type 1 and type 2 diabetic patients. RESEARCH DESIGN AND METHODS: Measurements were made on 15 type 1 (duration 13-32 years) and on 50 recently diagnosed type 2 diabetic patients (duration 3-4 years). The vasoconstrictor responses in the distal phalanx of the middle finger (locally heated to 40 degrees C) to the cooling of the contralateral arm were measured using Laser Doppler Imaging (LDI). A vasoconstriction index (VAC) was calculated taking age into account and was compared with reference values obtained in 80 control subjects. The diabetic patients were also studied with deep-breathing tests (i.e., the heart-rate variation expressed as the expiration-to-inspiration [E/I] ratio, a test of parasympathetic nerve function). RESULTS: The vasoconstrictor responses to indirect cooling (VAC) were significantly reduced in the fingers of the diabetic patients, both type 2 (0.77 +/- 0.02 V; P < 0.01) and type 1 (0.83 +/- 0.04 V; P < 0.001), compared with the healthy control subjects (0.65 +/- 0.01); the age-corrected VAC (VACz) was slightly more impaired in type 1 than in type 2 diabetic patients. The frequency of an abnormal VACz corresponded well to the frequency of an abnormal E/I ratio in type 1 diabetic patients (approximately 50%), whereas the frequency of an abnormal VACz was significantly higher than an abnormal E/I ratio among type 2 diabetic patients (11/50 vs. 4/50; P < 0.05). CONCLUSIONS: Both type 1 and type 2 diabetic patients have impaired cutaneous blood flow regulation. The VAC index seems to be a promising tool for detection of subclinical changes in autonomic sympathetic function.

Role of Growth arrest-specific gene 6-Mer axis in multiple myeloma.

Multiple myeloma is a mostly incurable malignancy characterized by the expansion of a malignant plasma cell (PC) clone in the human bone marrow (BM). Myeloma cells closely interact with the BM stroma, which secretes soluble factors that foster myeloma progression and therapy resistance. Growth arrest-specific gene 6 (Gas6) is produced by BM-derived stroma cells and can promote malignancy. However, the role of Gas6 and its receptors Axl, Tyro3 and Mer (TAM receptors) in myeloma is unknown. We therefore investigated their expression in myeloma cell lines and in the BM of myeloma patients and healthy donors. Gas6 showed increased expression in sorted BMPCs of myeloma patients compared with healthy controls. The fraction of Mer(+) BMPCs was increased in myeloma patients in comparison with healthy controls whereas Axl and Tyro3 were not expressed by BMPCs in the majority of patients. Downregulation of Gas6 and Mer inhibited the proliferation of different myeloma cell lines, whereas knocking down Axl or Tyro3 had no effect. Inhibition of the Gas6 receptor Mer or therapeutic targeting of Gas6 by warfarin reduced myeloma burden and improved survival in a systemic model of myeloma. Thus, the Gas6-Mer axis represents a novel candidate for therapeutic intervention in this incurable malignancy.

Design, synthesis, and biological characterization of bivalent 1-methyl-1,2,5,6-tetrahydropyridyl-1,2,5-thiadiazole derivatives as selective muscarinic agonists.

Selective muscarinic agonists could be useful in the treatment of neurological disorders such as Alzheimer's disease, schizophrenia, and chronic pain. Many muscarinic agonists have been developed, yet most exhibit at best limited functional selectivity for a given receptor subtype perhaps because of the high degree of sequence homology within the putative binding site, which appears to be buried within the transmembrane domains. Bivalent compounds containing essentially two agonist pharmacophores within the same molecule were synthesized and tested for receptor binding affinity and muscarinic agonist activity. A series of bis-1,2,5-thiadiazole derivatives of 1,2,5,6-tetrahydropyridine linked by an alkyloxy moiety exhibited very high affinity (K(i) < 1 nM) and strong agonist activity. The degree of activity depended on the length of the linking alkyl group, which could be replaced by a poly(ethylene glycol) moiety, resulting in improved water solubility, binding affinity, and agonist potency.

Plasma adiponectin and serum advanced glycated end-products increase and plasma lipid concentrations decrease with increasing duration of type 2 diabetes.

OBJECTIVE: To prospectively follow the concentrations of plasma adiponectin (p-adiponectin) and serum advanced glycation end-products (s-AGE) in relation to plasma lipids and retinopathy over 3 years in type 2 diabetic patients. DESIGN AND METHODS: P-adiponectin, s-AGE, plasma lipids and diabetic retinopathy were prospectively evaluated in 61 type 2 diabetic patients at baseline and at follow up 3 years later. RESULTS: Mean p-adiponectin (from 8.84+/-5.14 to 11.05+/-6.16 microg/ml; P=0.006) and s-AGE (from 637+/-242 to 781+/-173 ng/ml; P<0.0001) concentrations had increased at follow up. In addition, HbA1c (7.7+/-1.7 to 7.4+/-1.4%; P=0.0045) and fasting C-peptide (1.00+/-0.38 to 0.81+/-0.35 nM; P=0.019) had decreased and all lipid variables had significantly improved at follow up. P-adiponectin correlated inversely with fasting C-peptide (r(s)=-0.273; P=0.045) and low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio (r(s)=-0.362; P=0.011), and directly with plasma HDL cholesterol (r(s)=0.381; P=0.005) at follow up. Analysis of variance with adiponectin and s-AGE as dependent variables and fasting C-peptide, plasma HDL and plasma LDL cholesterol as covariates demonstrated that the increase in s-AGE was independent (P=0.001) and the increase in p-adiponectin dependent on covariate changes (P=0.862). There was a slight correlation between s-AGE at baseline versus the degree of retinopathy at follow up (r(s)=0.281; P=0.0499). CONCLUSION: Both p-adiponectin and s-AGE increased during the 3 years. The increase in p-adiponectin was explained by improvements in insulin sensitivity and dyslipidaemia, whereas the increase in s-AGE was independent of changes in metabolic covariates. s-AGE increase when the duration of type 2 diabetes increases.

Therapeutic effects of antibodies against adhesion molecules in murine collagen type II-induced arthritis.

Adhesion molecules play important roles in immune reactions and inflammatory processes and may constitute attractive targets for immunomodulatory approaches. In this study, blocking mAbs against a series of adhesion molecules were tested for their therapeutic effect on developing arthritis in a mouse model. MAbs were given for a period of 4 weeks at the time of exspected incidence of visible disease symptoms, i.e. 4 weeks after priming with collagen type II. A significant reduction of incidence down to values of 13% and 29% of the controls was obtained with mAbs against CD44 and alpha 4-integrin, respectively, during an observation time of 13 weeks. MAbs against CD4 and LFA-1 resulted only in weaker, non-significant effects or a delay in the incidence. MAbs against other molecules including L-selectin, ICAM-1 or VCAM-1 were not effective. The development of antibodies against collagen type II, collagen type I, proteoglycans and the immunogen, bovine collagen type II was affected by mAb treatment to a different extent. In this case, the anti CD4 mAb was the most effective, followed by the anti alpha 4-antibodies in most cases, whereas anti CD44 showed less clear effects on the development of humoral responses. In a skin delayed type hypersensitivity model analyzed for comparison, mAbs against LFA-1/ICAM-1 and alpha 4-integrin showed the largest effects on ear swelling. These data show that mAbs against several adhesion molecules are able to block selectively distinct aspects of immune reactions, and that CD44 and alpha 4-integrins could be promising targets for an immunotherapy of rheumatoid arthritis with receptor-interfering agents.

Impending electrical shock can affect response force in a simple reaction task.

For 20 subjects reaction times and force of response were measured on a simple reaction time task to visual stimuli while activation was manipulated by occasionally delivering a noninformative electrical shock. In blocks in which shocks were delivered, forces of response were larger than those in control blocks without shocks. The results are discussed in terms of Sanders' model of stress.

Nurses gain more time with patients.

Clinical pathways that incorporate charting by exception eliminate repetitive documentation and give nurses more time to educate and care for patients. In this case, nurses report a gain of 15 minutes per patient each day.

[Susceptibility to antibiotics of some species of Enterobacter isolated from various clinical materials]. / Wrazliwosc na antybiotyki niektórych gatunków rodzaju Enterobacter izolowanych z róznych materialów klinicznych.

We investigated 185 strains of Enterobacter spp. (E. cloacae--143 strains, E. agglomerans--15 strains, E. sakazakii--14 strains and E. aerogenes--13 strains) isolated from several clinical materials. An agar diffusion assay on Mueller-Hinton Agar was performed (according to NCCLS procedure and National Reference Center of Microorganisms' Drug-sensitiveness at Central Laboratory of Sera and Vaccinations in Warsaw Poland). E. cloacae: about 90% of the strains were sensitive to carbapenems. 85.3% of the isolates were susceptible to piperacillin + tazobactam. About 80% of the strains were susceptible to cotrimoxazole and ciprofloxacin. E. agglomerans: all strains were susceptible to imipenem, and 86.7%--to meropenem. 86.7% of the strains were susceptible to ureidopenicillins + inhibitors of beta-lactamases. About 80% of the isolates were susceptible to cotrimoxazole and ciprofloxacin. E. sakazakii: all isolates were susceptible to carbapenems. About 90% of the strains were susceptible to cotrimoxazole and 50%--to ciprofloxacin. E. aerogenes: all of the strains were susceptible to carbapenems and ciprofloxacin, 92.3%--to cotrimoxazole.

Cement mantle stress under retroversion torque at heel-strike.

The paper presents a theory of fixation failure and loosening in cemented total hip prostheses and proceeds to investigate this using an experimentally validated finite element model and two prosthesis types, namely the Charnley and the C-Stem. The study investigates the effects of retroversion torque occurring at heel-strike in combination with a loss of proximal cement/bone support and distal implant/cement support with a good distal cement/bone interface. A 3D finite element model was validated by comparison of femoral surface strains with those measured in an in vitro experimental simulation using an implanted Sawbone femur loaded in the heel-strike position and including a simplified representation of muscle forces. Results showed that the heel-strike position applies a high retroversion torque to the femoral stem that when combined with proximal debonding of the cement/bone interface and distal debonding of the implant/cement interface increases the strain transfer to the cement that may ultimately lead to the breakdown of the cement mantle leading on to osteolysis and loosening of the prostheses. Experimental fatigue testing of the implanted Charnley stem in a Sawbone femur produced cracks within the cement mantle that were located in positions of maximum stress supporting the finite element analysis results and theory of failure.

Potential biomarkers of an exaggerated response to endotoxemia.

Serial plasma protein analysis was used to study the acute plasma proteome response to endotoxemia (presence of toxic bacterial products called endotoxins in the blood stream). Plasma samples from healthy volunteers before and multiple time points up to 24 h following administration of low-dose endotoxin were evaluated. Plasma protein profiles were obtained by rapid extraction of whole plasma followed by analysis with matrix-assisted laser desorption ionisation-time of flight mass spectrometry. The profiles were unique to each individual and stable over the time of the experiment. Administration of low-dose endotoxin caused profound change in six of 18 individuals. At 8 h many proteins showed quantitative oxidation, in addition to the appearance of new components and disappearance of common baseline components. An exceptionally intense new component at 4154 mass units was identified as the activation peptide of C1 esterase inhibitor. While recovery of baseline protein structure was nearly complete by 24 h, serum amyloid A, an acute-phase reactant, was still increasing and minor profile changes persisted. Clinical features did not distinguish these extreme responders from others, suggesting that plasma proteome changes offered unique insights into and potential biomarkers of subclinical events following endotoxin exposure.

Peptic ulcer in geriatric long-term care medicine.

This study was based upon deceased patients in a geriatric university hospital with a high autopsy rate (81%). Of 6200 autopsied patients, 333 (5.4%) had had an active peptic ulcer; agonal and other acute erosions were not included. 257 cases were selected for the study (average age 83.8 yr). The diagnostic accuracy, and the symptoms of peptic ulcer in stationary, elderly, chronically ill patients were studied retrospectively. Only 16% of cases with duodenal ulcer and 29% with gastric ulcer had been correctly diagnosed antemortem. The clinical features of ulcer disease in the elderly may often differ from the standard presentation in younger people. Prior to death, appetite and weight loss, nausea/vomiting, anaemia and positive occult blood test had been more common among patients with ulcer, than abdominal pain and heartburn. The predictive values of single symptoms and of combined findings were low (range 2-21%), thus supporting observations from clinical practice that diagnosis is difficult in geriatric medicine. Prospective studies of ulcer disease in living elderly are needed.

Peritonitis in geriatric inpatients.

Of 212 cases of peritonitis found in a retrospective study of geriatric inpatients, the most common causes were mesenteric infarction, malignancy, intestinal obstruction, perforated peptic ulcer, cholecystitis, diverticulitis and perforation of the urinary bladder. The diagnostic accuracy was 47%. Abdominal pain had been observed in only 55% of the cases, and guarding and/or abdominal rigidity in only 34%. Other findings such as tachycardia and fever were more common, but the specificities of these signs were low.

Ulcer disease among geriatric inpatients with positive faecal occult blood test and/or iron deficiency anaemia. A prospective study.

The purpose of this study was to investigate the prevalence and type of lesions in the upper gastrointestinal tract and to identify characteristics associated with ulcer disease among geriatric inpatients with positive faecal occult blood test and/or iron deficiency anaemia. Two thousand five hundred and four patients aged 60-98 (mean, 82) years admitted to a geriatric clinic for rehabilitation were screened by faecal occult blood test, for B-haemoglobin, and, in a case of anaemia, analyses of serum levels of mean corpuscular volume, mean corpuscular haemoglobin concentration, iron, and total iron-binding capacity. One hundred and seventy patients were included in the study. A high prevalence of ulcer disease (22%) was found. Significantly higher proportions of non-steroidal anti-inflammatory drugs and steroid users and of patients with rheumatoid arthritis and osteoarthrosis were found among ulcer patients than among patients without ulcerative upper gastrointestinal lesions. The clinical picture of ulcer disease differed from the classic presentation: abdominal pain occurred in only 7 of 38 patients (18%), whereas appetite and weight loss and nausea/vomiting were common. It is important to be aware of the high prevalence and the clinical picture of ulcer disease among geriatric inpatients with iron deficiency anaemia and/or occult gastrointestinal bleeding.