Agrin induced morphological and structural changes in growth cones of cultured hippocampal neurons.

The role of agrin in synaptogenesis has been extensively studied. On the other hand, little is known about the function of this extracellular matrix protein during developmental processes that precede the formation of synapses. Recently, agrin was shown to regulate the rate of neurite elongation and the behavior of growth cones in hippocampal and spinal neurons, respectively. However, the molecular mechanisms underlying these effects have not been completely elucidated. In the present study, we analyzed the morphological and molecular changes induced by agrin in growth cones of hippocampal neurons that developed in culture. Morphometric analysis showed a significant enlargement of growth cones of hippocampal neurons cultured in the presence of agrin. These agrin-induced growth cone changes were accompanied by the formation of loops of microtubules highly enriched in acetylated tubulin and an increase in the content of the microtubule-associated protein (MAP)1B. Together, these data provide further insights into the potential molecular mechanisms underlying the effects of agrin on neurite outgrowth in rat central neurons.

Progesterone-induced agrin expression in astrocytes modulates glia-neuron interactions leading to synapse formation.

Experimental evidence recently obtained suggests that synaptogenesis is a tripartite event in which not only pre- and post-synaptic neurons but also glial cells play a key role. However, the molecular mechanisms by which glia modulate the formation of synapses in the CNS remain poorly understood. In the present study, we analyzed the role of astrocytes in synapse formation in cultured hippocampal rat neurons. For these experiments, hippocampal neurons were cultured in the presence or absence of a monolayer of astrocytes. Our results indicated that hippocampal neurons cultured in the presence of astrocytes formed more synapses than the ones cultured in their absence only when kept in N2 serum-free medium. To get insights into the potential molecular mechanisms underlying this effect, we analyzed the expression of proteins known to induce synapse formation in hippocampal neurons. A significant increase in agrin expression was detected in astrocytes cultured in N2 serum-free medium when compared with the ones cultured in serum containing medium. Experiments performed using different components of the N2 mixture indicated that progesterone induced the expression of agrin in astrocytes. Taken collectively, these results provide evidence supporting a role for astrocytes in synapse formation in central neurons. Furthermore, they identified agrin as a potential mediator of this effect, and astrocytes as a bridge between the endocrine and nervous systems during synaptogenesis.

Concentration of nutrients from urine and reject water from anaerobically digested sludge.

Experiments with concentration of nutrients from source separated urine and reject water from digestion of sludge in sewage treatment plants (STP) have been performed in laboratory and pilot scale. The methods tested were membrane filtration with reverse osmosis (RO), evaporation, and precipitation of phosphorus and distillation of ammonia. In membrane filtration, pre-filtration with particle separation at 5-10 microm was enough to avoid clogging of the membranes. Separation of phosphorus (P), potassium (K) and sulphur (S) was almost 100%, while separation of nitrogen (N) was dependent on pH. The capacity of flux increased with temperature and pressure. In evaporation, all P, K and S were still in the concentrate, while pH had to be decreased to 4.5 to avoid significant loss of N. In precipitation and distillation, about 90% of P could be recovered from urine as magnesium ammonium phosphate (MAP) just by adding MgO. For the reject water pH was first increased by aeration to remove CO2. Ammonium can be distilled from the water phase after precipitation of MAP, without further increase of pH. At least 80-90% of N can be distilled in 5-10% of the total volume. The article also discusses the quality of different products, cost of separation, and energy and chemical demand.

Prevalence trends of obesity and overweight among 10-year-old children in western Sweden and relationship with parental body mass index.

AIM: To determine the regional prevalence, secular and family-related trends of obesity and overweight among 10-y-old children. METHODS: A cross-sectional study of 10-y-old children, born in 1990, was performed during September 2000 to June 2001 at school health centres in three communities in the western part of Sweden. Evaluation was performed in 6311 children, or 81% of the target population. Data from a cohort of children, born in 1974, who form the national growth charts, were available for comparison. RESULTS: The mean body mass index was 17.9 kg/m2 in 10-y-old children born in 1990 and 17.0 kg/m2 for 10-y-olds born in 1974 (p < 0.0001). Of the 10-y-old children in 2000-2001, born in 1990, 18% were overweight and 2.9 % obese, which corresponds to a twofold increase in presence of overweight and a fourfold increase in presence of obesity among 10-y-old children from 1984 to 2000. There was a significant correlation between parental and child body mass index. The prevalence of obesity and being overweight appeared to be higher in children whose parents did not participate in the study. CONCLUSION: During a 16-y period, from 1984 to 2000, a twofold increase in being overweight and a fourfold increase in obesity were seen among 10-y-old children in the western part of Sweden. Parental ponderosity or reluctance to participate in the study was related to a higher prevalence of being overweight or obese in the children. There is a need for the healthcare system to recognize the threats to the health of the population of this new "epidemic" and initiate preventive measures and treatment programmes.

Increasing incidence rates of squamous cell carcinoma of the skin in Sweden.

The incidence of squamous cell carcinoma of the skin is increasing world-wide, and in Sweden this tumour is one of the most rapidly increasing malignancies. The aim of this study was to investigate incidence trends of squamous cell carcinoma in Sweden. For the 39,805 tumours registered in the Swedish Cancer Registry 1961-1995, incidence rates were calculated according to gender, age, anatomical site and unit surface area. Multivariate analysis was performed with the age-period-cohort model. Age-standardized incidence rates increased substantially in both men (+425%) and women (+146%) during this period. The highest rates per unit surface area were seen for chronically sun-exposed head-neck sites. Age-specific incidence rates increased in ages > or =60 years during the study period. Multivariate analyses showed that age, period and cohort effects in men could best explain the incidence rates, while in women the age-period effects model was adequate. In conclusion, a rapidly increasing incidence trend for squamous cell carcinoma was found, probably explained by increased accumulated sun exposure and increasing incidence among the elderly.

The structure of a Michaelis serpin-protease complex.

Serine protease inhibitors (serpins) regulate the activities of circulating proteases. Serpins inhibit proteases by acylating the serine hydroxyl at their active sites. Before deacylation and complete proteolysis of the serpin can occur, massive conformational changes are triggered in the serpin while maintaining the covalent linkage between the protease and serpin. Here we report the structure of a serpin-trypsin Michaelis complex, which we visualized by using the S195A trypsin mutant to prevent covalent complex formation. This encounter complex reveals a more extensive interaction surface than that present in small inhibitor-protease complexes and is a template for modeling other serpin-protease pairs. Mutations of several serpin residues at the interface reduced the inhibitory activity of the serpin. The serine residue C-terminal to the scissile peptide bond is found in a closer than usual interaction with His 57 at the active site of trypsin.

Trends in lung cancer incidence in Sweden with special reference to period and birth cohorts.

OBJECTIVE: Sweden has one of the largest population-based cancer registers in the world that provides an opportunity to examine the trend of lung cancer incidence during a 35-year period. The primary aim of the present study was to estimate the effects of birth cohort, year of diagnosis (period), and age on the time trends of lung cancer incidence rates, and to analyze the gender-specific incidence of different histopathological types of lung cancer. RESULTS: Among men the age-standardized incidence rate increased steadily up to 1982, when a peak of 49 cases per 100,000 person-years was reached. Among women the incidence rate was lower and showed a monotonic increase throughout the observation period. The fastest rate of increase was noted among the youngest women. In women, but not in men, there was a steady increase in risk with each successive birth cohort. For both sexes there were large changes in the histopathological distributions of cases. The most notable was a major increase in adenocarcinomas. CONCLUSIONS: The overall age-adjusted incidence rate of lung cancer in Sweden has stabilized in men during the past two decades while rates are still increasing in women. In view of the continued high prevalence of smoking among young women, a future definite increase in the overall number of lung cancer cases in women can be expected.

A bridge between the RNA and protein worlds? Accelerating delivery of chemical reactivity to RNA and DNA by a specific short peptide (AAKK)(4).

BACKGROUND: RNA can catalyze diverse chemical reactions, leading to the hypothesis that an RNA world existed early in evolution. Today, however, catalysis by naturally occurring RNAs is rare and most chemical transformations within cells require proteins. This has led to interest in the design of small peptides capable of catalyzing chemical transformations. RESULTS: We demonstrate that a short lysine-rich peptide (AAKK)(4) can deliver a nucleophile to DNA or RNA and amplify the rate of chemical modification by up to 3400-fold. We also tested similar peptides that contain ornithine or arginine in place of lysine, peptides with altered stereochemistry or orientation, and peptides containing eight lysines but with different spacing. Surprisingly, these similar peptides function much less well, suggesting that specific combinations of amino acids, charge distribution, and stereochemistry are necessary for the rate enhancement by (AAKK)(4). CONCLUSIONS: By appending other reactive groups to (AAKK)(4) it should be possible to greatly expand the potential for small peptides to directly catalyze modification of DNA or RNA or to act as cofactors to promote ribozyme catalysis.

Mouse models for the Wolf-Hirschhorn deletion syndrome.

Wolf-Hirschhorn syndrome (WHS) is a deletion syndrome caused by segmental haploidy of chromosome 4p16.3. Its hallmark features include a 'Greek warrior helmet' facial appearance, mental retardation, various midline defects and seizures. The WHS critical region (WHSCR) lies between the Huntington's disease gene, HD, and FGFR3. In mice, the homologs of these genes map to chromosome 5 in a region of conserved synteny with human 4p16.3. To derive mouse models of WHS and map genes responsible for subphenotypes of the syndrome, five mouse lines bearing radiation-induced deletions spanning the WHSCR syntenic region were generated and characterized. Similar to WHS patients, these animals were growth-retarded, were susceptible to seizures and showed midline (palate closure, tail kinks), craniofacial and ocular anomalies (colobomas, corneal opacities). Other phenotypes included cerebellar hypoplasia and a shortened cerebral cortex. Expression of WHS-like traits was variable and influenced by strain background and deletion size. These mice represent the first animal models for WHS. This collection of nested chromosomal deletions will be useful for mapping and identifying loci responsible for the various subphenotypes of WHS, and provides a paradigm for the dissection of other deletion syndromes using the mouse.

Risk for gastric cancer after antibiotic prophylaxis in patients undergoing hip replacement.

Despite strong evidence of an association between Helicobacter pylori and gastric cancer, the benefit of eradicating H. pylori infection is unknown. Our aim was to test the hypothesis that exposure to high doses of antibiotics reduces risk for gastric cancer via possible eradication of H. pylori We conducted a nationwide case-control study nested in a cohort of 39,154 patients who underwent hip replacement surgery between 1965 and 1983. Such patients frequently receive prophylactic antibiotic treatment. During follow-up through 1989, we identified 189 incident cases of gastric cancer. For each case, three controls were selected from the cohort. Exposure data were abstracted from hospital records. Blood samples from a separate cohort undergoing hip replacement surgery were analyzed for anti-H. pylori IgG before and after surgery. Both long-term antibiotic treatment before surgery [odds ratio (OR), 0.3; 95% confidence interval (CI), 0.1-0.7] and prophylactic antibiotic treatment (OR, 0.7; 95% CI, 0.5-1.1) conferred a reduction in gastric cancer risk. The reduction appeared stronger after 5 years (OR, 0.6; 95% CI, 0.3-1.2) than during shorter follow-up after hip replacement (OR, 0.8; 95% CI, 0.4-1.7). There was an apparent decrease in risk with increasing body weight-adjusted doses of antibiotics (P = 0.13). However, the rate of H. pylori antibody disappearance was not strikingly higher in the cohort of patients undergoing hip replacement than in a control cohort. Our findings provide indirect support for the hypothesis that treatment with antibiotics at a relatively advanced age reduces the risk of gastric cancer.

Surgery for stomach cancer in a defined Swedish population: current practices and operative results. Swedish Gastric Cancer Study Group.

OBJECTIVE: To study routine surgical in-hospital care, and to relate postoperative morbidity and mortality to age, sex, tumour stage, operation done, and surgical workload of the hospital. DESIGN: Prospective population-based study. SETTING: All hospitals that diagnosed new cases of stomach cancer in five counties in central and northern Sweden, 1 February 1989-31 January 1995. PATIENTS: All 1024 patients diagnosed as having a new adenocarcinoma of the stomach. RESULTS: The stomach cancer was in such an advanced stage at diagnosis that only half of the patients could be offered a potentially curative operation. The tumour was resectable in 632 patients (62%). Distal gastric resection was done for 359 (57%) and total gastrectomy in 259 (41%) of all the resected cases. Postoperative complications occurred in 250 patients (31%). In multivariate analyses the relative risk (RR) for postoperative complications increased to 2.3 (95% confidence interval [CI] = 1.3 to 4.3) in patients over 79 years of age compared with those under 60. The corresponding RR for postoperative death was 5.1 (95% CI = 2.0 to 12.7) in patients over 79 years. Total gastrectomy combined with splenectomy and distal pancreatectomy carried the highest postoperative morbidity (RR = 3.3) and mortality (RR = 3.7) compared with distal gastrectomy. CONCLUSION: There was no difference in postoperative morbidity or mortality among different types of hospital categories. Surgical treatment of stomach cancer still carries a substantial morbidity and mortality in an unselected series of patients, particularly among elderly patients.

Type 2 diabetes and the metabolic syndrome in Japanese Americans.

Japanese Americans have experienced a higher prevalence of type 2 diabetes than in Japan. Research conducted in Seattle suggests that lifestyle factors associated with 'westernization' play a role in bringing out this susceptibility to diabetes. These lifestyle factors include consumption of a diet higher in saturated fat and reduced physical activity. A consequence of this is the development of central (visceral) adiposity, insulin resistance, and other features associated with this insulin resistance metabolic syndrome, such as dyslipidemia (high triglycerides, low HDL-cholesterol, and small and dense LDL particles), hypertension, and coronary heart disease. We have postulated that the superimposition of insulin resistance upon a genetic background of reduced beta-cell reserve results in hyperglycemia and diabetes among Japanese Americans. This article reviews evidence that support this view.

A multivariate analysis of factors affecting patency of femoropopliteal and femorodistal bypass grafting.

BACKGROUND: The most important factors that determine the outcome after femoropopliteal and femorodistal arterial reconstruction are still controversial. This report analysis the factors that determine the early and late patency of distal arterial reconstruction. PATIENTS AND METHODS: A retrospective analysis of patency after femorodistal arterial reconstruction with a new method for evaluation of angiographic runoff was performed for 336 arterial reconstructions. The different pre-, per- and postoperative risk factors were analysed in a Cox proportional hazards model. RESULT: The patency was significantly better for vein grafts in comparison to composite grafts and prosthetic grafts. It was 74% for vein, 46% for composite and 43% for prosthetic reconstructions, respectively, at 12 months after arterial reconstruction. The cumulative life table patency rate in extremities with good, intermediate and poor runoff was 62, 30 and 10%, respectively at 36 months. The patency rates for extremities operated on for claudication was significantly better than for extremities operated on for critical ischaemia. The multivariate analysis of different factors in a Cox analysis revealed that only the status of distal runoff, the graft material and the site of the distal anastomosis independently and significantly influenced the patency rates. CONCLUSIONS: A new model for evaluation of distal runoff proved to predict the patency rate of femoropopliteal and femorodistal arterial reconstructions reasonably well in this retrospective analysis.

Factors affecting reproducibility of dietary reports using food frequency questionnaires.

OBJECTIVE: To study the reproducibility of a food frequency questionnaire used to assess past dietary habits. DESIGN: Repeated retrospective assessment of dietary habits of a population sample. SETTING: Uppsala-Orebro Health Care Region of Sweden. SUBJECTS: One-hundred and four subjects participating as control subjects in a case-control study on thyroid cancer. INTERVENTIONS: Between spring 1993 and spring 1994 a case-control study was conducted in the study area, in order to investigate risk factors for thyroid cancer-diet among others. The study subjects reported their consumption of selected foods with reference 1-5 y before, as well as dietary changes that occurred from adolescence to adult age. One year later 197 control persons were invited to provide a second report of the same food items. Of the invited subjects 104 agreed to participate. RESULTS: The median Spearman rank correlation coefficient between the first and second assessment was 0.58 for food consumption and 0. 56 for nutrients intake. Correlation between the two assessments varied greatly between food items, ranging from -0.03 to 0.83. The correlation was positively related to the frequency and negatively related to the skewness of consumption. The recall of dietary changes from adolescence had a median correlation coefficient of 0. 38 (range 0.19-0.69). There were hints of a higher reproducibility of dietary reports among men and among highly educated subjects (more than 11 y of education), but these differences were statistically significant only for recall of adolescent diet. Reproducibility of nutrients intake was also significantly higher among men than among women, as well as among subjects older than 45 y. CONCLUSIONS: The reproducibility of dietary reports was satisfactory. Among factors affecting reproducibility, frequency and homogeneity of consumption in the source population are presumably the most important. Age, sex and education modify the inter-subject reproducibility of past diet. Based on reproducibility of a semi-quantitative index, the recall of adolescent diet probably has a low validity. European Journal of Clinical Nutrition (2000) 54, 658-664.

Cholecystectomy, peptic ulcer disease and the risk of adenocarcinoma of the oesophagus and gastric cardia.

BACKGROUND: Gastro-oesophageal reflux is a strong risk factor for oesophageal adenocarcinoma. Bile and pancreatic enzymes may be particularly carcinogenic. Cholecystectomy causes an increased gastric level of these constituents. A decreased risk of oesophageal adenocarcinoma has been observed in persons infected with cagA-positive Helicobacter pylori. There is a strong correlation between ulcer disease and Helicobacter pylori infection. The aim of this study was to determine whether previous cholecystectomy or peptic ulcer disease affects the risk of oesophageal carcinoma. METHODS: Data were collected as a nationwide population-based case-control study in Sweden between 1995 and 1997. Multivariate adjusted odds ratios (ORs) were calculated with logistic regression. RESULTS: There was no statistically significant association between cholecystectomy and the risk of oesophageal carcinoma. Among persons with previous peptic ulcer, the adjusted OR for oesophageal adenocarcinoma was below unity (OR = 0.6, 95 per cent confidence interval 0.3-1.1). The relative risk estimates for cardia adenocarcinoma and oesophageal squamous cell carcinoma were close to unity. CONCLUSION: Cholecystectomy, despite its effect on the composition of gastric juice, does not appear to increase the risk of adenocarcinoma of the oesophagus or gastric cardia. While the data do not contradict a protective effect of H. pylori, the results are also consistent with absence of such an effect.

Decreasing incidence of both major histologic subtypes of gastric adenocarcinoma--a population-based study in Sweden.

While the overall incidence of gastric cancer has fallen, presumably to a large extent in parallel with Helicobacter pylori infection, the occurrence of the diffuse histologic type is thought to have remained more stable, questioning the aetiologic role of H. pylori. We have analysed the incidence of the intestinal and diffuse types separately, while considering subsite (cardia/non-cardia). With an extensive prospective effort we identified all incident cases of gastric adenocarcinoma (n = 1337) in a well-defined Swedish population (1.3 million) 1989-1994. Tumours were uniformly classified histologically and topographically. Subgroup-specific incidence rates were computed and modelled using multivariate logistic regression. Site-specific trends were clearly discrepant. The overall incidence of adenocarcinoma distal to the gastric cardia declined by 9% (95% confidence interval 6-12%) per year, while cardia cancer remained stable. Thus, the feared rise in cardia cancer could not be confirmed despite clear site-specific trend discrepancies. The intestinal type predominated, especially in high-risk areas, while diffuse tumours prevailed among young patients and women. Both main histologic types of gastric adenocarcinoma declined markedly, at similar rapidity, and with no significant trend differences between the intestinal and diffuse types, even after multivariate adjustments. Our results are consistent with an aetiologic role of environmental factors including H. pylori also for diffuse-type gastric cancers.

Association between medications that relax the lower esophageal sphincter and risk for esophageal adenocarcinoma.

BACKGROUND: The incidence of esophageal adenocarcinoma is increasing rapidly. Gastroesophageal reflux is a strong risk factor for this disease. The increase in incidence of esophageal adenocarcinoma coincided with the introduction of medications that promote reflux by relaxing the lower esophageal sphincter (LES), such as nitroglycerin, anticholinergics, beta-adrenergic agonists, aminophyllines, and benzodiazepines. OBJECTIVE: To test the possible association between use of LES-relaxing medications and risk for adenocarcinoma of the esophagus and gastric cardia. DESIGN: A nationwide population-based case-control study with in-person interviews. SETTING: Sweden, 1995 through 1997. PATIENTS: 189 patients with newly diagnosed esophageal adenocarcinoma, 262 with adenocarcinoma of the gastric cardia, and 167 with esophageal squamous-cell carcinoma were compared with 820 population-based controls. MEASUREMENTS: Estimated incidence rate ratios, calculated by using multivariate logistic regression from case-control data with adjustment for potential confounding. RESULTS: Past use of LES-relaxing drugs was positively associated with risk for esophageal adenocarcinoma. Among daily, long-term users (>5 years) of LES-relaxing drugs, the estimated incidence rate ratio was 3.8 (95% CI, 2.2 to 6.4) compared with persons who had never used these drugs. Drugs of all classes contributed to the increased risk, but the association was particularly strong for anticholinergics. Short-term use of other types of LES-relaxing drugs did not seem to be strongly associated with risk. The association almost disappeared after adjustment for reflux symptoms, indicating that promotion of reflux is the link between use of LES-relaxing drugs and esophageal adenocarcinoma. If 15,490 men in any age group take LES-relaxing drugs daily for 5 years, 1 additional case of adenocarcinoma would be expected (number needed to treat for harm); in men older than 60 years of age, the number needed to treat for harm is 5,570. Assuming a causal relation, about 10% of the esophageal adenocarcinomas occurring in the population may be attributable to intake of LES-relaxing drugs. Cardia adenocarcinoma and esophageal squamous-cell carcinoma were not associated with use of LES-relaxing drugs. CONCLUSIONS: The widespread use of LES-relaxing drugs may have contributed to the increasing incidence of esophageal adenocarcinoma.

Interdigitated deletion complexes on mouse chromosome 5 induced by irradiation of embryonic stem cells.

Chromosome deletions have several applications in the genetic analysis of complex organisms. They can be used as reagents in region-directed mutagenesis, for mapping of simple or complex traits, or to identify biological consequences of segmental haploidy, the latter being relevant to human contiguous gene syndromes and imprinting. We have generated three deletion complexes in ES (Embryonic Stem) cells that collectively span approximately 40 cM of proximal mouse chromosome 5. The deletion complexes were produced by irradiation of F(1) hybrid ES cells containing herpes simplex virus thymidine kinase genes (tk) integrated at the Dpp6, Hdh (Huntington disease locus), or Gabrb1 loci, followed by selection for tk-deficient clones. Deletions centered at the adjacent Hdh and Dpp6 loci ranged up to approximately 20 cM or more in length and overlapped in an interdigitated fashion. However, the interval between Hdh and Gabrb1 appeared to contain a locus haploinsufficient for ES cell viability, thereby preventing deletions of either complex from overlapping. In some cases, the deletions resolved the order of markers that were previously genetically inseparable. A subset of the ES cell-bearing deletions was injected into blastocysts to generate germline chimeras and establish lines of mice segregating the deletion chromosomes. At least 11 of the 26 lines injected were capable of producing germline chimeras. In general, those that failed to undergo germline transmission bore deletions larger than the germline-competent clones, suggesting that certain regions of chromosome 5 contain haploinsufficient developmental genes, and/or that overall embryonic viability is cumulatively decreased as more genes are rendered hemizygous. Mice bearing deletions presumably spanning the semidominant hammertoe locus (Hm) had no phenotype, suggesting that the classic allele is a dominant, gain-of-function mutation. Overlapping deletion complexes generated in the fashion described in this report will be useful as multipurpose genetic tools and in systematic functional mapping of the mouse genome.