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Objective Biomarkers for diagnosis and progression of osteoarthritis (OA) are lacking. This study was undertaken to identify circulating biomarkers for OA that could predict disease occurrence and/or progression to joint replacement. Methods Using the SomaScan platform, we measured 4,792 proteins in plasma from 37,278 individuals, of whom 12,178 individuals had OA and 2,524 had undergone joint replacement. We performed a case-control study for identification of potential protein biomarkers for hip, knee, and/or hand OA, and a prospective study for identification of biomarkers for joint replacement. Results Among the large panel of plasma proteins assessed, cartilage acidic protein 1 (CRTAC1) was the most strongly associated with both OA diagnosis (odds ratio 1.46 [95% confidence interval 1.41-1.52] for knee OA, odds ratio 1.36 [95% confidence interval 1.29-1.43] for hip OA, and odds ratio 1.33 [95% confidence interval 1.26-1.40] for hand OA) and progression to joint replacement (hazard ratio 1.40 [95% confidence interval 1.30-1.51] for knee replacement and hazard ratio 1.31 [95% confidence interval 1.19-1.45] for hip replacement). Patients with OA who were in the highest quintile of risk of joint replacement, based on known risk factors (i.e., age, sex, and body mass index) and plasma CRTAC1 level, were 16 times more likely to undergo knee replacement within 5 years of plasma sample collection than those in the lowest quintile, and 6.5 times more likely to undergo hip replacement. CRTAC1 was not associated with other types of inflammatory arthritis. A specific protein profile was identified in those patients who had undergone joint replacement prior to plasma sample collection. Conclusion Through a hypothesis-free approach, we identified CRTAC1 in plasma as a novel promising candidate biomarker for OA that is both associated with occurrence of OA and predictive of progression to joint replacement. This biomarker might also be useful in the selection of suitable patients for clinical trial enrollment.

The authors have withdrawn this manuscript because this paper was posted prematurely in advance of a UK Biobank Pharma Proteomics Project consortium effort. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author

The plasma proteome can help bridge the gap between the genome and diseases. Here we describe genome-wide association studies (GWASs) of plasma protein levels measured with 4,907 aptamers in 35,559 Icelanders. We found 18,084 associations between sequence variants and levels of proteins in plasma (protein quantitative trait loci; pQTL), of which 19% were with rare variants (minor allele frequency (MAF)

About 5-8% of individuals over the age of 60 have dementia. With our ever-aging population this number is likely to increase, making dementia one of the most important threats to public health in the 21st century. Given the phenotypic overlap of individual dementias the diagnosis of dementia is a major clinical challenge, even with current gold standard diagnostic approaches. However, it has been shown that certain dementias show specific structural characteristics in the brain. Progressive supranuclear palsy (PSP) and multiple system atrophy (MSA) are prototypical examples of this phenomenon, as they often present with characteristic brainstem atrophy. More detailed characterization of brain atrophy due to individual diseases is urgently required to select biomarkers and therapeutic targets that are meaningful to each disease. Here we present a joint multi-atlas-segmentation and deep-learning-based segmentation method for fast and robust parcellation of the brainstem into its four sub-structures, i.e., the midbrain, pons, medulla, and superior cerebellar peduncles (SCP), that in turn can provide detailed volumetric information on the brainstem sub-structures affected in PSP and MSA. The method may also benefit other neurodegenerative diseases, such as Parkinson's disease; a condition which is often considered in the differential diagnosis of PSP and MSA. Comparison with state-of-the-art labeling techniques evaluated on ground truth manual segmentations demonstrate that our method is significantly faster than prior methods as well as showing improvement in labeling the brainstem indicating that this strategy may be a viable option to provide a better characterization of the brainstem atrophy seen in PSP and MSA., Accepted to SPIE Medical Imaging 2021

Publisher's version (útgefin grein). Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations., Discovery of coding variants in genes that confer risk of neurodevelopmental disorders is an important step towards understanding the pathophysiology of these disorders. Wholegenome sequencing of 31,463 Icelanders uncovers a frameshift variant (E712KfsTer10) in microtubule-associated protein 1B (MAP1B) that associates with ID/low IQ in a large pedigree (genome-wide corrected P = 0.022). Additional stop-gain variants in MAP1B (E1032Ter and R1664Ter) validate the association with ID and IQ. Carriers have 24% less white matter (WM) volume (β = −2.1SD, P = 5.1 × 10−8), 47% less corpus callosum (CC) volume (β = −2.4SD, P = 5.5 × 10−10) and lower brain-wide fractional anisotropy (P = 6.7 × 10−4). In summary, we show that loss of MAP1B function affects general cognitive ability through a profound, brain-wide WM deficit with likely disordered or compromised axons., We are grateful to the participants and we thank the psychologists, nurses and staff, in particular Berglind Eiriksdottir, at the Research Recruitment Center and technicians and staff at Röntgen Domus. We also thank the staff at deCODE genetics core facilities and all our colleagues for their important contribution to this work. L.J. received support from the Swedish Society of Medicine, the Swedish Brain Foundation and Swedish Society for Medical Research. The research leading to these results has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreements’ no. 115008 (NEWMEDS) and no. 115300 (EUAIMS) of which resources are composed of EFPIA in-kind contribution and financial contribution from the European Union’s Seventh Framework Programme (EU-FP7/2007-2013), EU-FP7 funded grant no. 602450 (IMAGEMEND) and EU funded FP7-People-2011-IAPP grant agreement no. 286213 (PsychDPC).

Introduction: Ammonia production from the metabolism of urea by urease enzymes of oral bacteria moderates plaque acidification and may inhibit dental caries, as suggested by in vitro studies and indirect clinical observations. The objective of this study was to examine the relationship of urease activity with dental caries at the clinical level. Methods: Urease activity was measured in dental plaque and saliva samples from 25 caries-free subjects (CF) and in eight subjects with six or more open caries lesions (CA). Plaque and saliva collection was repeated for each subject 1 week later using identical procedures. Results: Urease-specific activity in the dental plaque of CF subjects was significantly higher compared to that in the subjects with caries. The association of low plaque urease levels with increased caries was further supported by odds ratio analysis using different plaque urease cut-off points. Using a receiver operating characteristic curve it was estimated that there was an approximately 85% probability of correctly classifying the subjects as CA or CF based on the relative ordering of their plaque urease activity levels. No statistically significant differences were observed in salivary urease activity. Conclusion: This study suggests that loss of alkali-generating potential of tooth biofilms via the urease pathway has a positive relationship to dental caries.

Pouchitis is the major long-term complication of restorative proctocolectomy for ulcerative colitis (UC). Allopurinol is a scavenger of oxygen-derived free radicals, which it is suggested play a role in the development of UC and pouchitis. The first aim was to test the hypothesis that the incidence of pouchitis can be reduced by prophylactic Allopurinol, and secondly to evaluate if Allopurinol influences the overall pouch function.273 patients with UC who were planned for proctocolectomy and ileal pouch-anal anastomosis at 12 centres in Sweden between October 1994 and June 1997 were offered the opportunity to participate. 184 patients (67%) were randomized to receive postoperative prophylactic Allopurinol 100 mg twice daily or placebo. All 273 patients had clinical and endoscopic follow-up at 1, 3, 6, 12, 18, 24 months after surgery.Of the 184 randomized patients, 94 were randomized to Allopurinol and 90 to placebo; 116 patients (63%) completed follow-up and the crude incidence of pouchitis among those patients fullfilling the protocol was 31% in the Allopurinol group and 28% in the placebo group (ns). The cumulative risk for a first attack of pouchitis was 30% and 26% after 24 months (ns). The overall pouch function improved over time and did not differ significantly between the two groups.Prophylactic Allopurinol did not reduce the risk of a first attack of pouchitis.

A detailed chemical model describing the formation of soot and NO is applied to simulate emission formation in a heavy duty diesel engine. Cylinder flow and spray development is simulated using an engine CFD code - Speedstar. Combustion is described using a simple eddy break-up model. Modeling of the emission-chemistry/turbulent-flow interaction is based on a flamelet approach. Contrary to a typical flamelet concept, transport equations are solved for mass fractions of soot and NO. The reason being that these major emission constituencies are assumed to change slowly in comparison to typical time scales for chemical processes or transport processes important for combustion. Chemical reactions leading to production and destruction of soot and NO are, however, assumed to be fast. Soot and NO source terms are therefore evaluated from a flamelet library using a presumed probability density function and integrating over mixture fraction space. Results from simulations are compared to engine measurements inform of exhaust emission data and cylinder pressure.

With the aid of specific monoclonal antibodies, an immunohistochemical technique has recently been developed for the detection of intratumoral thymidylate synthase (TS). This technique can be applied to paraffin-embedded material suitable for retrospective studies. In order to examine this technique further, the TS enzyme activity of lysates from frozen-stored colorectal cancer (CRC) specimens were compared with their immunohistochemical TS staining intensity (arbitrarily graded from 0 to 3). A statistically significant correlation between these two methods on a total of 25 tumour specimens (P< 0.001) was observed. The staining intensity in different areas of4S paraffin-embedded CRCs was examined. Sixty-seven per cent of the tumours were homogeneously stained (either grades 0–1 or 2–3), 33% showed a heterogeneity in TS staining. Increased TS expression correlated with more advanced Dukes' stage (P< 0.001). It is concluded that TS immunostaining intensity reflects TS enzyme activity in colorectal tumours and is well suited for paraffin-embedded material. The TS immunostaining pattern is heterogeneous in up to one-third of the tumours.

We designed and performed a multicenter clinical trial to determine the relationship between measurements of the level of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) to other measures used to detect periodontal disease and monitor outcome of treatment, including pocket depth and gingival inflammation. 32 periodontitis patients were enrolled at the University of Washington, Seattle, 30 at the University of Florida, Gainesville, and 34 at the University of Illinois, Chicago. 10 periodontally normal control subjects were enrolled at each location. 8 diseased and 4 healthy sites were designated for study in each patient and 8 healthy sites designated in each control subject. Measures of disease included pocket depth, severity of gingival inflammation, and GCF volume. AST levels were measured using the PerioGard test kit. Clinical measurements were made and GCF samples harvested and tested 2x before and 2x after therapy consisting of scaling and root planing under local anesthetic. Specific design and other issues are discussed, including selection of patients and control subjects, sample size, selection of experimental test sites, methods for assessment of diseased and therapeutic improvement, harvesting of GCF and selection of appropriate biostatistical methods for data analysis. Demographics of the patient populations at the 3 locations are reported. As expected, therapy induced only negligible changes in the measures of disease at healthy sites in control subjects, and relatively minor improvement in healthy sites in patients. In contrast, statistically significant improvement relative to pretreatment baseline status in all 3 measures of disease was observed for diseased sites at all 3 study locations with all p-values less than 0.0002. The magnitude of improvement was comparable to that reported previously by others. The % of PerioGard-positive sites decreased significantly between the screening baseline and both post-treatment visits for patients at all 3 locations, with p values of 0.0001 to

Purposes of this study were 1) to estimate in humans, using 14C-labeled propionate, the rate of hepatic gluconeogenesis relative to the rate of Krebs cycle flux; 2) to compare those rates with estimates previously made using [3-14C]lactate and [2-14C]acetate; 3) to determine if the amount of ATP required for that rate of gluconeogenesis could be generated in liver, calculated from that rate of Krebs cycle flux and splanchnic balance measurements, previously made, and 4) to test whether hepatic succinyl-CoA is channeled during its metabolism through the Krebs cycle. [2-14C]propionate, [3-14C]-propionate, and [2,3-14C]succinate were given along with phenyl acetate to normal subjects, fasted 60 h. Distributions of 14C were determined in the carbons of blood glucose and of glutamate from excreted phenylacetylglutamine. Corrections to the distributions for 14CO2 fixation were made from the specific activities of urinary urea and the specific activities in glucose, glutamate, and urea previously found on administering [14C]-bicarbonate. Uncertainties in the corrections and in the contributions of pyruvate and Cori cyclings limit the quantitations. The rate of gluconeogenesis appears to be two or more times the rate of Krebs cycle flux and pyruvate's decarboxylation to acetyl-CoA, metabolized in the cycle, less than one-twenty-fifth the rate of its decarboxylation. Such estimates were previously made using [3-14C]lactate. The findings support the use of phenyl acetate to sample hepatic alpha-ketoglutarate. Ratios of specific activities of glucose to glutamate and glucose to urinary urea and expired CO2 indicate succinate's extensive metabolism when presented in trace amounts to liver. Utilizations of the labeled compounds by liver relative to other tissues were in the order succinate = lactate > propionate > acetate. ATP required for gluconeogenesis and urea formation was approximately 40% of the amount of ATP generated in liver. There was no channeling of succinyl-CoA in the Krebs cycle in the hepatic mitochondria.

Attachment level has been used as the "gold standard" for assessment of the progression of periodontal disease. However, the measurement of attachment level by periodontal probing can be subject to a large number of error sources. Recently, we have designed experiments by using an electronic probe to identify the magnitude of error components due to the instrument, gingival tissue condition, position or probing angle, and time interval between replicate probings. Even with a very careful clinical setting, a few percent of uncontrollable large errors or outliers could not be avoided. A previously used 'option-3' probing scheme to reduce the unexpected large error is justified from the mathematical viewpoint.

To examine the fate of the carbons of acetate and to evaluate the usefulness of labeled acetate in assessing intrahepatic metabolic processes during gluconeogenesis, [2-14C]acetate, [2-14C]ethanol, and [1-14C]ethanol were infused into normal subjects fasted 60 h and given phenyl acetate. Distributions of 14C in the carbons of blood glucose and glutamate from urinary phenylacetylglutamine were determined. With [2-14C]acetate and [2-14C]ethanol, carbon 1 of glucose had about twice as much 14C as carbon 3. Carbon 2 of glutamate had about twice as much 14C as carbon 1 and one-half to one-third as much as carbon 4. There was only a small amount in carbon 5. These distributions are incompatible with the metabolism of [2-14C]acetate being primarily in liver. Therefore, [2-14C]acetate cannot be used to study Krebs cycle metabolism in liver and in relationship to gluconeogenesis, as has been done. The distributions can be explained by: (a) fixation of 14CO2 from [2-14C]acetate in the formation of the 14C-labeled glucose and glutamate in liver and (b) the formation of 14C-labeled glutamate in a second site, proposed to be muscle. [1,3-14C]Acetone formation from the [2-14C]acetate does not contribute to the distributions, as evidenced by the absence of 14C in carbons 2-4 of glutamate after [1-14C]ethanol administration.

A mouse monoclonal antibody (MAb17-1A) (IgG2A) against colorectal carcinoma cells was used to treat patients with metastatic disease. Major direct effector functions of MAb seem to be ADCC (antibody dependent cellular cytotoxicity), CDC (complement dependent cytolysis) and apoptosis ('programmed cell death'). Thus, a high tumor cell saturation of the MAb should be achieved. Increasing doses of MAb to the patients increased the total area under the concentration curve and thus the exposure of tumor cells to MAb. However, the response rate (with complete + partial + minor response + stable disease defined as response) was not augmented. In total, 10/52 (19%) patients responded and in fact lower doses (less than 2 g) might induce a higher response frequency (9/52) than higher doses (greater than 2 g) (1/52). During treatment, the numbers of cytotoxic cells (lymphocytes and monocytes) increases in the tumor lesion and complement components were deposited. As ADCC may be important, effector mechanism attempts were made to augment the cytolytic capability of the effector cells by simultaneously giving the patients GM-CSF. The combination of MAb17-1A + GM-CSF augmented the ADCC activity of blood mononuclear cells and a heavy infiltration of monocytes could be noted in the tumor. Out of 15 available patients 6 (40%) showed a response.

Two new methods to assess liver metabolism in humans will be discussed, non-invasive probes and 13C NMR (nuclear magnetic resonance) spectroscopy. Hepatic UDP-glucose and α-ketoglutarate can be sampled by non-invasive probes. This method has been used to quantitate the contribution of the indirect pathway to liver glycogen formation and to estimate relative flow rates and carbon exchange in the Krebs cycle. 13C NMR spectroscopy has been developed to measure liver glycogen concentrations in vivo. Using this method in combination with measurements of whole body glucose production during fasting, the contribution of gluconeogenesis has been calculated.

Several studies have suggested that the intratumoral level of thymidylate synthase (TS) in colorectal tumors correlates with survival. We have studied the correlation between TS expression in primary rectal cancer and locoregional recurrence, distant metastases, and survival. TS enzyme levels were evaluated immunohistochemically using the specific monoclonal antibody TS 106 in paraffin-embedded tumors from 243 patients who had undergone primary surgery for rectal cancer during the years 1980-1993. All patients were included in prospective randomized trials aimed at determining the clinical value of a short preoperative course of local radiation therapy (five doses of 5 Gy each). With a median follow-up of 94 months (range, 43-202 months), it was observed by multivariate analysis that Dukes' stage and TS expression were independent prognostic markers of locoregional recurrence (P0.001 and P = 0.038, respectively) distant metastasis (P0.001 and P = 0.011, respectively) disease-free survival (P0.001 and 0.014, respectively), and overall survival (P0.001 and 0.020, respectively). By multivariate analysis, preoperative irradiation therapy showed a borderline improvement in locoregional recurrence (P = 0.051). No other factors, such as age, sex, differentiation of the tumor, or p53 expression, were noted to be independent prognostic factors for clinical outcome in these patients. We concluded that the intratumoral expression of TS in primary rectal cancer is an independent prognostic factor for locoregional recurrence, distant metastases, disease-free survival, and overall survival. Patients with low intratumoral TS expression had a significantly better outcome than those with high TS expression.

Intratumoral thymidylate synthase (TS) expression and M(r) 53,000 phosphoprotein (p53) overexpression were studied immunohistochemically in sections from stored paraffin-embedded primary colorectal cancers in 70 patients who had undergone surgery during the years 1987-1990. These cancers were classified according to Dukes' stage A-D, using monoclonal antibodies TS 106 and DO-7. In patients with Dukes' stage A-C tumors, univariate analyses showed that there was a significant correlation (P = 0.048) between disease-free survival and TS expression and between TS expression and time to death with colorectal cancer (P = 0.038). In patients with Dukes' stage A-D tumors, overall survival was correlated to TS expression (P = 0.015), Dukes' stage (P0.001), and level of tumor differentiation (P = 0.044) but not to p53 overexpression. Patients with low intratumoral TS expression survived significantly longer than patients with high expression. Cox multivariate analysis showed that Dukes' stage (P0.001) and TS expression (P = 0.043) could independently serve as prognostic factors for time to death with colorectal cancer in patients with Dukes' stage A-D tumors.

Periodontitis is a result of an infection with specific pathogenic microorganisms. Thus, the local delivery of antimicrobials has been investigated as a possible method for controlling this infection and treating periodontal disease. A number of antimicrobial agents have been studied both as adjunctive therapies with scaling and root planing and as stand-alone chemotherapies. These agents have been administered in irrigation solutions and as single-dose formulations, but with little long-term efficacy in the treatment of periodontitis. Recent investigations have focused on the delivery of antimicrobials in sustained-release formulations designed to maintain effective concentrations of drug within the periodontal pocket. This article provides an overview of the development of the use of locally delivered antimicrobials in periodontal therapy and the current state-of-the-art of the technique.

Periodontal disease in the elderly has not been characterized. Initial reports suggest that the disease is common and severe. Deficiencies in the immune response have also been reported to occur in the elderly. Consequently it was hypothesized that aging-related changes in the immune response may contribute to the severity and occurrence of periodontal disease in the elderly. To test that hypothesis, the % and number of leukocytes and leukocyte subsets in the peripheral blood of elderly (65-75 years) subjects were tested and used as indicators of the immune potential of those individuals. Age and gender effects on several of the parameters tested were identified. With the exception of increased number of leukocytes in the elderly group with severe periodontal disease, no other alteration in the leukocyte parameters tested were identified. These results suggest that periodontal disease in the elderly was not associated with obvious changes in the leukocyte and leukocyte subsets in the peripheral blood due to aging.

Digital subtraction radiography (DSR) has been shown to be a sensitive and specific method for the detection of small bony changes in periodontitis. The purpose of this study was to perform a multicenter validation of the DSR in human subjects. Seventeen subjects were enrolled at 3 centers. Feather-edged hydroxyapatite chips (approximately 1, 7 and 10 mg) were used to simulate osseous lesions. Bilateral radiographs were taken with and without chips. Geometry was standardized using a cephalostat and the order of radiographs was determined using a randomization plan. Radiographs were subtracted, lesions isolated, and quantified at a single center without knowledge of the randomization code or location of the chips used in each subject. The overall sensitivity and specificity in detecting 1 mg changes was 87.8% and 100%, respectively. Sensitivity and specificity in detecting 7 mg and 10 mg chips was 100%. A strong linear relationship between actual lesion mass and calculated mass was observed (R2 = 0.94, slope = 0.98, p < 0.0001). No significant differences were observed by center. These data indicate that the DSR is a valid technique for the assessment of alveolar bone changes in multicenter trials.

We have previously reported a preferential usage of V delta 1/V gamma 8 on TCR-gamma-delta-bearing intraepithelial lymphocytes of the normal human intestine as well as in the inflamed synovial tissue of patients with rheumatoid arthritis. The aim of the present study was to analyze V gene segment usage by gamma delta T cells of the intestine and peripheral blood (PB) from patients with inflammatory bowel disease. Freshly isolated lymphocytes were analyzed by flow cytometry using a panel of V gene subset-specific mAbs. The relative proportion of PB TCR-gamma delta+ cells was increased in patients with Crohn's disease as compared with controls. Interestingly, an increased proportion of PB gamma delta T cells of patients with ulcerative colitis or CrD expressed V delta and V gamma genes typically used by intraepithelial lymphocytes. Thus, increased proportions of V delta 1+ and V gamma 8+ cells were found in the PB of both patient groups. The majority of TCR-gamma delta+ intraepithelial lymphocytes (IEL) in inflamed and noninflamed intestine expressed V delta 1/V gamma 8, and a substantial proportion of V gamma(2,3,4)+ cells were found. These observations suggest a disease-associated appearance of "gut-like" gamma delta T cells in the periphery. Moreover, two patients had a large proportion of gamma delta T cells in their PB of which the majority expressed two distinct V gamma proteins. Both of these patients had a short duration of disease (1 and 10 mo, respectively) and the relative proportion of gamma delta T cells decreased in concert with stabilization of disease. Interestingly, one of the patients had a clonal expansion of a V gamma dual-expressing gamma delta T cell in the PB.

Subcutaneous administration of low doses of recombinant interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) on an out-patient basis has been reported not to significantly compromise the response frequency compared to intravenous IL-2 in patients with renal cell carcinoma and melanoma. As part of an ongoing program to develop a biotherapeutic concept in patients with colorectal carcinoma (CRC) we studied the clinical effects of such a regimen in 15 patients with metastatic CRC. The daily dose of IL-2 varied between 4.8-14.4 x 10(6) U/m2 and of IFN-alpha between 3-6 x 10(6) U/m2. The cycle length was 6 weeks. The course was repeated every 8 weeks until disease progression. Maximum 4 cycles were administered. Maintenance therapy was given to responding patients once a week every month. No patient showed a major response (CR or PR). Six patients had a stable disease ranging from 3 months to 18 months with a median duration time of 5 months. The median survival of all patients was 13 months. The main adverse reactions were fever, chills, anorexia and shortness of breath. No treatment related deaths occurred. 6/14 patients developed abnormal concentration of serum levels of thyroid hormones. It is concluded that the present treatment schedule using IL-2 and IFN-alpha in advanced CRC seemed not to be of clinical benefit.

Three different 5-fluorouracil (5-FU)-interferon-α-2b (IFN)-containing regimens were designed for treatment of patients with advanced colorectal cancer. 87 patients with a Karnofsky index ≥70 were included in three sequential non-randomised phase II trials. Regimen A consisted of 5-FU (750 mg/m 2 /day) given as a continuous infusion on days 1–5 followed by weekly 1-h intravenous infusions until week 8. IFN (5 MU) was given subcutaneously on days 1, 3 and 5 followed by injections (9 MU) every second day until week 8. The cycle was then repeated. Regimen B consisted of 5-FU (750 mg/m 2 /day) given as a continuous infusion on days 1–5 followed by 5-min intravenous injections on days 12 and 19. IFN (3 MU) was given subcutaneously on days 1–5 followed by injections (5 MU) on days 11–13 and 18–20. The cycle was repeated every fourth week. Regimen C consisted of 5-FU (750 mg/m 2 /day) given as a continuous infusion on days 1–5. IFN (3 MU) was given subcutaneously on days 1–5. The cycle was repeated every third week. The objective response rates (complete response (CR) and partial response (PR)) after approximately 4 months of therapy or longer were as follows: regimen A ( n = 27) 22% (2 CR, 4 PR), regimen B ( n = 33) 42% (4 CR, 10 PR) and regimen C ( n = 27) 22% (1 CR, 5 PR). The corresponding response figures for previously untreated patients were regimen A 50%, regimen B 64% and regimen C 38%. Response durations varied from a few weeks up to 142 + weeks. Toxicities were generally mild and reversible, and the treatments were convenient for the patients and cost effective since the 5-day infusions could be given by a portable pump without hospitalisation. Our results are in agreement with those of others showing that 5- FU IFN combinations can be highly effective in advanced colorectal cancer, and that a number of factors such as doses, dose intensities, infusion rates and timing of the two drugs may be crucial for the anti-tumour activity of this drug combination.

The intestinal population of gamma delta T cell receptor (TCR)-bearing cells was characterized with regard to V delta and V gamma subtype expression. For this purpose, we utilized V gene-specific PCR of mRNA prepared from intestinal biopsies. Predominant expression of the V delta 1 subtype was demonstrated in the small intestine of patients with coeliac disease and in the inflamed colon of patients with inflammatory bowel diseases (IBD: ulcerative colitis and Crohn's disease) as well as in colon biopsies taken from macroscopically normal areas of colon. Although intestinal gamma delta T cells preferentially expressed V delta 1, other V delta transcripts could be detected, of which V delta 2 and V delta 5 were commonly expressed. Analysis of biopsies from mesenteric lymph nodes demonstrated a V delta repertoire similar to the mucosa. In peripheral blood on the other hand, high expression of both V delta 2 and V delta 1 was found. The predominant expression of V delta 1 transcripts in the intestinal mucosa of IBD patients correlated well with protein cell surface expression as analysed by flow cytometry using V delta 1- and V delta 2-specific antibodies. Selective expansion of gamma delta T cells could not be demonstrated within the inflamed mucosa as shown by mRNA analysis and flow cytometry. Instead, IBD patients demonstrated a decreased proportion of TCR gamma delta-carrying T cells in the inflamed mucosa compared with macroscopically normal area of colon. On the other hand, a significantly increased percentage of T cells bearing the gamma delta TCR was found in peripheral blood of patients with Crohn's disease compared with healthy individuals, indicating that local mucosal inflammation may influence the circulating gamma delta T cell population.

An impaired immune response is frequently observed in cancer patients and tumor-bearing mice. T-cells from mice with an experimental colon carcinoma were recently shown to express T-cell receptors that completely lacked the signal-transducing molecule CD3 zeta. Here, we have investigated the expression of the signal-transducing molecule zeta on lymphocytes from 14 patients with colorectal carcinomas using flow cytometric analysis of permeabilized cells with a monoclonal antibody (TIA-2; IgG1) specific for the cytoplasmic domain of the zeta chain as well as with immunoprecipitation and analysis on diagonal gel electrophoresis. We demonstrate that T-cells isolated from the tumors of the patients express significantly less CD3 zeta than T-cells in the peripheral blood of the same patients and that the peripheral blood of the patients express decreased levels of zeta chains, as compared to the levels found in lymphocytes from healthy controls. This decreased expression was also observed on zeta chains associated with the low affinity Fc receptor for IgG found in tumor-infiltrating NK cells (Fc gamma RIIIA alpha; CD16).

Unconjugated monoclonal antibodies (MAb) and granulocyte macrophage-colony stimulating factor (GM-CSF) may induce tumor regression in patients. Antibody-dependent cellular cytotoxicity (ADCC) is considered to be one of the effector functions of MAb. Human peripheral blood mononuclear cells (PBMC) preincubated with GM-CSF and used as effector cells in an 18h ADCC assay with SW948 (human colorectal carcinoma cell line) as target cells and MAb 17-1A induced significant increase in the lytic capacity of the effector cells. Based on these findings the therapeutic effect of the combination of mouse MAb 17-1A (IgG2a) against colorectal carcinoma (CRC) cells and GM-CSF was evaluated in 20 patients with metastatic CRC. The patients received GM-CSF (250 micrograms/m2/day s.c.) for 10 days and a single i.v. infusion of MAb 17-1A (400 mg) at day 3 of the cycle. The cycles were repeated with an interval of one month. Four cycles were given. ADCC as well as Fc-receptor bearing mononuclear cells increased significantly during therapy. Two patients achieved CR (10%). One patient had an MR (5%) and a further three patients were considered to have SD > 3 months (15%). The two CR patients are still in CR, 35+ and 30+ months respectively after initiation of therapy. Patients with an ADCC activity at start of therapy above the median value of the total patient material survived significantly longer than those patients with an ADCC reactivity below this value (p = 0.002).

To estimate the relative contribution of the indirect pathway to liver glycogen formation in humans, different experimental approaches have been used. Although the estimates vary, it appears that in overnight fasted humans approximately 50% of liver glycogen is derived from the direct pathway following a glucose load and that the direct pathway increases to approximately 70% after the first meal of the day. While the source of the gluconeogenic substrates is still unknown, the animal data suggest that the liver itself may be an important source.

To examine the fate of the carbons of acetate and to evaluate the usefulness of labeled acetate in assessing intrahepatic metabolic processes during gluconeogenesis, [2-14C]acetate, [2-14C]ethanol, and [1-14C]ethanol were infused into normal subjects fasted 60 h and given phenyl acetate. Distributions of 14C in the carbons of blood glucose and glutamate from urinary phenylacetylglutamine were determined. With [2-14C]acetate and [2-14C]ethanol, carbon 1 of glucose had about twice as much 14C as carbon 3. Carbon 2 of glutamate had about twice as much 14C as carbon 1 and one-half to one-third as much as carbon 4. There was only a small amount in carbon 5. These distributions are incompatible with the metabolism of [2-14C]acetate being primarily in liver. Therefore, [2-14C]acetate cannot be used to study Krebs cycle metabolism in liver and in relationship to gluconeogenesis, as has been done. The distributions can be explained by: (a) fixation of 14CO2 from [2-14C]acetate in the formation of the 14C-labeled glucose and glutamate in liver and (b) the formation of 14C-labeled glutamate in a second site, proposed to be muscle. [1,3-14C]Acetone formation from the [2-14C]acetate does not contribute to the distributions, as evidenced by the absence of 14C in carbons 2-4 of glutamate after [1-14C]ethanol administration.

To quantify intrahepatic Krebs cycle metabolism, phenyl acetate, excreted in urine as a glutamine conjugate, was given to healthy subjects infused with [3-14C]lactate. They were studied after 60 h of fasting and when given glucose after an overnight fast. Distributions of 14C in glutamate from urinary phenylacetylglutamine and blood glucose were determined. Corrections to the distributions because of the fixation of 14CO2 formed from the [3-14C]lactate were determined by administering [14C]bicarbonate. Comparisons of distributions in glucose and glutamate support the assumption that the glutamate distributions reflect those in hepatic alpha-ketoglutarate. From the distributions in glutamate, the extent of exchange of labeled with unlabeled carbons and relative flow rates in the cycle in liver were estimated. Dilution of 14C by 12C in the cycle was found in the fasted but not the fed state. In the fasted state, pyruvate carboxylation was estimated to be at least twice the rate of Krebs cycle flux and the rate of pyruvate's decarboxylation less than 1/25 the rate of its carboxylation. In the fed state, the rate of decarboxylation was estimated to be between one-sixth and one-half the rate of carboxylation. The rate of conversion of oxalacetate to fumarate in both states appeared to be greater than 6 times the rate of Krebs cycle flux.

The haemostatic effect of terlipressin (triglycyl-lysine vasopressin; Glypressin) on bleeding from oesophageal varices was evaluated in a placebo-controlled, double-blind, randomized clinical trial. Patients with clinically suspected liver cirrhosis were included in the study if they had been admitted to hospital with an extensive haemorrhage within the last 24h before diagnostic endoscopy. The patients randomized after stratification for severity of liver disease. Terlipressin or placebo was administered as intravenous bolus injections every 4th h during a period of 24 to 36 h or until the clinical course necessitated active intervention (failure or withdrawal). Sixty patients entered the study; 31 patients were allocated to receive terlipressin, and 29 patients to receive placebo. Bleeding from varices was arrested in 28 of the 31 receiving terlipressin, as compared with 17 of the 29 receiving placebo (p less than 0.01). Patients receiving active drug required significantly fewer blood transfusions (p less than 0.05). Most of the side effects were classified as mild and were registered in the terlipressin group.

Ischaemic colitis was not diagnosed until laparotomy in five cases. In two it was secondary to hypotension and multiple organ failure, in two it was atherosclerotic, and one had polycythemia vera. The diagnosis should always be considered in patients with abdominal distension--with or without diarrhoea and abdominal pain--so that treatment may be instituted before the onset of peritonitis.

In this paper we wish to demonstrate the multi-element capability of Laser Enhanced Ionization (LEI) in flames by reporting the detection of 15 different elements. All the elements were excited by the fundamental or the first harmonic frequency-doubled light from one single dye in the laser system (Coumarin 47). Detection limits below the ppb region (parts-per-billion, ng/ml) were achieved for 13 elements (Al, Bi, Ca, Cd, Co, Cr, Fe, Ga, Li, Ni, Pb, Sr, Tl). The detection limits reported here are for several elements orders of magnitude superiour to those for existing comparable flame based spectroscopic methods. For 9 elements, the detection limits are found to be lower than any previously reported values for one-step LEI in flames.

A graphite furnace is successfully used as an atomizer in laser-enhanced ionization spectroscopy (LEI). The physical origin of the signals is discussed and a comparison is made with LEI measurements in a flame. Extremely small amounts of analyte atoms can be detected using this method. The reproducibility is good, there are almost no memory effects, and the signal output is linear with the amount of analyte over at least 5 orders of magnitude. The experimental results indicate that the method could become an attractive alternative for trace-element analysis.

A theoretical model is developed which relates the experimentally observed signal to the charged species produced in laser enhanced ionization. The model is based on a general equation, which describes the electric field in a flame confined between electrodes at fixed potentials, and a relation for the charge induced on these electrodes. A complete analytical solution is found in a one-dimensional geometry. The signal corresponding to the laser induced electrons is described accurately by the model. The ionic part of the signal, although in most cases negligible, is influenced significantly by dynamic screening in the flame, and requires a more complete model. The theory is applied to an experimental situation, and relations for optimal signal collection are derived. Then the predictions from the theory in a cylindrical geometry are compared to results from a graphite furnace experiment. The temporal behaviour of the signal is further used for a determination of the electron mobility in a flame.

Earlier works on rabbits have demonstrated that destruction of both labyrinths leads to a defective OKN in acute stage. The aim of this study was to examine whether this defect is to be ascribed to changes in brain stem mechanisms of OKN or due to possible disturbances of alertness. In 10 intact rabbits and in 6 hemilabyrinthectomized rabbits (HL-rabbits) OKN and EEG were studied. Alertness was controlled with vibration. In intact rabbits OKN reached a maximum velocity slowly, during several minutes. Simultaneously, theta waves were observed in EEG indicating increase in brain activity. Exposure to vibration caused a sooner increase and higher a maximum velocity of OKN and a momentary increase in EEG activity, than without vibration stimuli. In HL-rabbits OKN remained reduced in both directions still two months after operation. Vibration only slightly improved OKN responses while EEG was normally activated. OKN was decreased to about the same extent as was vestibular nystagmus. Attention seems to influenc...

One-step Laser-Enhanced Ionization (LEI) spectrometry of 23 different elements in aqueous solutions has been performed in an acetylene/air flame. All elements were detected by light in the ultraviolet region, produced by frequency doubling of the output from the dye Coumarin 153. This was done in order to investigate the multielement capability of LEI in flames that has been made possible by the recent development of commercially available, widely tunable dyes. Among the elements detected, 9 (As, Au, In, Mn, Pb, Sb, Tl, W, Yb) show detection limits which are superior to those reported in the literature for one-step LEI. The lowest detection limit obtained in this investigation was 1 pg/mL for In. Four of the elements (As, Sb, Yb, W) are reported as being detected by LEI for the first time. The multielement capabilities of LEI as a method for trace element analysis are discussed.

Spectral interferences from matrix elements in solutions are investigated utilizing Laser Enhanced Ionization (LEI) in flames. Mechanisms which influence the analytical selectivity are discussed and it is shown that the selectivity can be significantly increased by a proper choise of experimental parameters. This is exemplified by determination of low concentrations of Mg in a matrix of Na. The spectral interference, occurring in one-step excitation LEI, is diminished by a factor of 70 when utilizing two-step excitation LEI. The spectral resolution is further improved to a total factor of 1300 by an optimized choice of laser light intensities without any significant loss in sensitivity. It is pointed out that an efficient correction for signal contributions from interfering substances can be made in two-step excitation LEI without scanning the laser wavelengths. Background signals emanating from NO molecules inherent to the flame gases are also detected and virtually eliminated. The very high sensitivity and selectivity achieved by two-step excitation LEI shows that the method is widely applicable for trace element analysis.

The aim of this study was to compare the periodontal probing depth in teeth exposed to periodontal treatment with teeth having received no treatment. In pockets deeper than 4 mm, a significant difference was found between the two groups. In the non-treated teeth, metal strips used for probing penetrated an average of 0.29 mm into the connective tissue apical to the junctional epithelium. The average probing depth for teeth exposed to scaling, root planing, curettage and chlorhexidine rinses for 1 month was 0.31 mm coronal of the connective tissue attachment within the junctional epithelium. The results indicate that care should be taken when evaluating changes in periodental probing measurements during longitudinal studies.

Various experimental conditions for laser-enhanced ionization (LEI) spectroscopy in a graphite furnace are investigated with the aim to find optimal conditions for trace element analysis. Some general aspects of the generation of LEI charges and the collection of the signal are discussed. It is established that many elements can be measured with very high sensitivity and good reproducibility. However, the determination of certain elements probably requires a modified experimental configuration. Such a modification is discussed in the paper.