[Immunoassay and discriminant analysis to improve the diagnosis of ovarian cancer in the elderly.]

Ovarian cancer is a common pathology in the structure of tumors among elderly women. The aim of the work is to identify the leading indicators of interleukin profile to improve the diagnosis of ovarian cancer in the elderly. Enzyme immunoassay studied serum levels of interleukins in 78 patients with ovarian cancer stage IIB-IIIB according to the classification of FIGO at the age of 65-70 years and 42 women of similar age without ovarian cancer at the time of the study and in history. The differentiation of the compared groups is carried out by means of discriminant method. The leading diagnostic interleukins for ovarian cancer in the elderly are IL-1α, IL-1ß, IL-8 and TNF-α. The constructed discriminant models allow to differentiate patients with ovarian cancer and control women with an accuracy of 93.2-93.8%. The use of enzyme immunoassay and discriminant analysis improve the diagnosis of ovarian cancer in the elderly.

[The application of flow cytometry technique and diagnostically significant indices of systemic cellular immunity for diagnostic of acute endometritis.]

The acute endometritis is a the most widespread pathology among women of reproductive age.The immune system plays a leading role in its development. However, there is no data concerning the most informative parameters of cell immunity. The purpose of study is to enhance diagnostic of acute endometritis using immunologic technique with objective evaluation of diagnostic value of indices of systemic cell immunity. The indices of cell immunity are analyzed on sampling of 78 patients with postnatal acute endometritis and 54 healthy women. The technique of flow cytometry was applied with subsequent calculation of index of informativeness and disintegration objectifying selection of diagnostically significant parameters. It is demonstrated that full-fledged diagnostic of acute endometritis requires detection of absolute and relative content of T-helpers and natural killers having high value of informativeness and disintegration. Hence an opportunity to reduce number of analyzed parameters of systemic cell immunity and financial costs of diagnostic of acute endometritis.

[The differentiated diagnostic of chronic salpingoophoritis and ovary cancer on the basis of immunologic and discriminant techniquea.]

The ovary cancer and chronic salpingoophoritis are among leading pathologies of female's sexual sphere and a main cause of morbidity and mortality of female population. The purpose of study was to select leading differential diagnostic indices of systemic cellular immunity and to carry out a differentiation of patients with ovary cancer and chronic salpingoophoritis using discriminant technique. The differentiated diagnostic of ovary cancer and chronic salpingoophoritis was applied to 92 and 87 patients correspondingly to indices of systemic cellular immunity and analyzed by flow cytometry technique. It is demonstrated that the main differential diagnostic indices of systemic cellular immunity in case of ovary cancer and chronic salpingoophoritis are decreasing of relative content of natural killers, absolute number of B-lymphocytes and T-helpers. The developed discriminant models permit to carry out in 91.8%-92.1% cases a faultless differentiated diagnostic of ovary cancer and chronic salpingoophoritis. The approach like that increases quality of differentiated diagnostic of ovary cancer and chronic salpingoophoritis.

Production and purification of lentiviral vectors generated in 293T suspension cells with baculoviral vectors.

Lentivirus can be engineered to be a highly potent vector for gene therapy applications. However, generation of clinical grade vectors in enough quantities for therapeutic use is still troublesome and limits the preclinical and clinical experiments. As a first step to solve this unmet need we recently introduced a baculovirus-based production system for lentiviral vector (LV) production using adherent cells. Herein, we have adapted and optimized the production of these vectors to a suspension cell culture system using recombinant baculoviruses delivering all elements required for a safe latest generation LV preparation. High-titer LV stocks were achieved in 293T cells grown in suspension. Produced viruses were accurately characterized and the functionality was also tested in vivo. Produced viruses were compared with viruses produced by calcium phosphate transfection method in adherent cells and polyethylenimine transfection method in suspension cells. Furthermore, a scalable and cost-effective capture purification step was developed based on a diethylaminoethyl monolithic column capable of removing most of the baculoviruses from the LV pool with 65% recovery.

Assays for acetaldehyde-derived adducts in blood proteins based on antibodies against acetaldehyde/lipoprotein condensates.

BACKGROUND: Acetaldehyde-derived protein condensates (adducts) have been suggested as promising biological markers of alcohol abuse because they represent actual metabolites of ethanol. However, the detection of such condensates in vivo has been hampered by a lack of sensitive and specific methods. METHODS: To develop new approaches for the detection of acetaldehyde adducts, we have raised antibodies against condensates with acetaldehyde and lipoproteins, which have previously been shown to be readily modified by acetaldehyde in vitro. The characteristics of these antibodies were compared with those raised against bovine serum albumin/acetaldehyde adduct and against other types of lipoprotein modifications, as induced by malondialdehyde, oxidation, and acetylation. RESULTS: The antibodies raised against low-density lipoprotein (LDL)/acetaldehyde, very low density lipoprotein (VLDL)/acetaldehyde, and bovine serum albumin/acetaldehyde all reacted with protein adducts generated at physiologically relevant concentrations of acetaldehyde in vitro, whereas the antibodies raised against malondialdehyde/LDL, oxidized LDL, or acetylated LDL were not found to cross-react with the acetaldehyde-derived adducts. In assays for acetaldehyde adducts from erythrocyte and serum proteins of patients with excessive ethanol consumption (n = 32) and healthy control individuals (n = 22), the antibody prepared against the acetaldehyde/VLDL condensate was found to provide the most effective detection of acetaldehyde adducts in vivo. CONCLUSIONS: Current data indicate that acetaldehyde generates immunogenic adducts with lipoproteins in vivo. Antibodies raised against the VLDL/acetaldehyde may provide a basis for new diagnostic assays to examine excessive alcohol consumption.

Fine root biomass and production in Scots pine stands in relation to stand age.

We determined fine root biomass and production of 15-, 35- and 100-year-old Scots pine (Pinus sylvestris L.) stands during three growing seasons. Fine roots were sampled by the soil core method. Mean (+/- SE) annual fine root biomass of Scots pine in the 15-, 35- and 100-year-old stands was 220 +/- 25, 357 +/- 21 and 259 +/- 26 g m(-2), respectively. Fine root biomass of the understory vegetation was 159 +/- 54 g m(-2), 244 +/- 30 and 408 +/- 81 g m(-2), and fine root necromass was 500 +/- 112, 1,047 +/- 452 and 1,895 +/- 607 g m(-2) in the sapling, pole stage and mature stands, respectively. Both understory and Scots pine fine root production increased with stand age. Mean annual Scots pine fine root production was 165 +/- 131, 775 +/- 339 and 860 +/- 348 g m(-2) year(-1) in the sapling, pole stage and mature stand, respectively. The respective mean annual production of all fine roots (Scots pine and understory) was 181 +/- 129, 1,039 +/- 497 and 1,360 +/- 869 g m(-2) year(-1). The Scots pine and understory fine root biomass, necromass and production varied in relation to stand age, although the variation was not statistically significant.

Serum IgG and IgE antibodies against mold-derived antigens in patients with symptoms of hypersensitivity.

BACKGROUND: Exposure to mold in water-damaged buildings has been suggested to be responsible for various health problems such as hypersensitivity and upper respiratory tract diseases. However, only little information is available on possible diagnostic tools for examining mold-associated health problems. METHODS: We used recently developed immunofluorometric IgG and IgE assays (UniCAP) to examine serum IgG and IgE antibodies against mold-derived allergens from 70 mold-exposed individuals with (n = 55) or without (n = 15) symptoms of sensitization. Controls were healthy individuals (n = 31) without any history of such exposure. RESULTS: The IgG titers exceeded the upper normal limits of control individuals (mean +/- 2 S.D.) in 35% of symptomatic men and in 25% of women. The IgG titers were usually higher in women than in men (P < 0.05) showing no significant association with the severity of symptoms. During follow-up of eight mold-exposed subjects for 9-12 months the IgG titers remained relatively constant. Elevated anti-mold IgEs were found in six (11%) of the exposed subjects who were all symptomatic. CONCLUSIONS: Measurements of anti-mold IgGs may help to confirm exposure in patients with hypersensitivity symptoms and evidence of mold growth in living or working environment. Some exposed symptomatic patients present IgE-mediated responses. Combined measurements of IgGs and IgEs may prove to be of value in the comprehensive assessment and treatment of such patients.

Autoimmune responses against oxidant stress and acetaldehyde-derived epitopes in human alcohol consumers.

BACKGROUND: Studies in experimental animals have indicated that chronic ethanol ingestion triggers the formation of antibodies directed against proteins modified with reactive metabolites of ethanol and products of lipid peroxidation. However, the nature and prevalence of such antibodies have not been compared previously in alcoholic patients. METHODS: Autoantibodies against adducts with acetaldehyde- (AA), malondialdehyde- (MDA), and oxidized epitopes (Ox) were examined from sera of 54 alcohol consumers with (n = 28) or without (n = 26) liver disease, and from 20 nondrinking controls. RESULTS: Anti-AA-adduct IgA and IgG antibodies were elevated in 64% and 31% of patients with biopsy-proven alcoholic liver disease (ALD, n = 28), respectively. The IgA titers were significantly higher than those from nondrinking controls (p < 0.001), or heavy drinkers without significant liver disease (p < 0.001). Anti-MDA adduct titers (IgG) were elevated in 70% of the ALD patients. These titers were significantly higher (p < 0.001) than those from nondrinking controls, or heavy drinkers without liver disease. Antibodies (IgG) against Ox epitopes occurred in 43% of ALD patients, and the titers also were significantly higher (p < 0.05) than those from nondrinking controls. The anti-AA and anti-MDA adduct titers in ALD patients correlated significantly with the combined clinical and laboratory index (CCLI) of liver disease severity (r(s) = 0.449, p < 0.05; r(s) = 0.566, p < 0.01, respectively), the highest prevalences of anti-AA-adducts (73%) and anti-MDA-adducts (76%) occurring in ALD patients with cirrhosis. CONCLUSIONS: The present results indicated that autoantibodies against several distinct types of protein modifications are generated in ALD patients showing an association with the severity of liver disease.

Alterations in airway wall properties in infants with a history of wheezing disorders.

Airway diameter and airway wall mechanics (compliance) are important determinants of flow limitation and wheezing. We have previously used the high-speed interrupter technique (HIT) to measure input impedance (Zin) in infants at frequencies up to 900 Hz, including antiresonance phenomena, which are known to be related to wave propagation velocity, and have shown that the frequency at which the first antiresonance occurs (f(ar,1)) is a function of airway wall compliance. We aimed to determine whether f(ar,1) (and thus airway wall compliance) was different in infants with a history of wheezing disorders. We compared 23 asymptomatic infants (aged 36 to 81 wk) with a history of wheezing with an age-matched group of 19 healthy control infants. We found that f(ar,1) was significantly lower in infants with wheezing disorders than in the control group (p < 0. 005), implying differences in airway wall compliance, even when they were clinically asymptomatic. Developmental differences in airway wall mechanics may be important in the pathogenesis of wheezing disorders or, alternatively, alterations in airway wall mechanics might be a consequence of postinflammatory remodeling.

No consistent pattern of mutations in p53 and ras genes in liver tumors of rat treated with the drinking water mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX).

The frequency of point mutations in p53 (exons 4-7) and in Ki-ras, Ha-ras, and N-ras (exons 1 and 2) and the expression of p53 protein were evaluated in the liver tumors of Wistar rats of a 104-week carcinogenicity study on 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), a chlorine disinfection by-product in drinking water. Mutations were analyzed in 16 hepatocellular adenomas, 7 hepatocellular carcinomas, 23 cholangiomas, and 2 cholangiocarcinomas of the MX-treated animals and one hepatocellular carcinoma and cholangiocarcinoma in control animals using PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) or PCR-TGGE (temperature gradient gel electrophoresis) and direct sequencing. The expression of the p53 protein (wild-type and mutated protein) was detected by immunohistochemistry (CM5 antibody). The expression of p53 and that of the proliferating cell nuclear antigen (PCNA, 19 A2) were also evaluated in livers of female animals exposed to MX for 1 week, 3 weeks, or 18 weeks. Altogether, four mutations were found in p53 in three tumors, in two hepatocellular adenomas, and one cholangiocarcinoma, all in females receiving the highest MX dose (6. 6 mg/kg/day) of the study. Three of the mutations were G:C --> A:T transitions and one was an A:T --> T:A transversion. The mutations were scattered at different codons and positions of the codon. One hepatocellular adenoma contained two p53 mutations. All cholangiomas and cholangiocarcinomas, but no hepatocellular adenomas and carcinomas, overexpressed the p53 protein. MX treatment did not induce p53 expression at any age in the liver or alter the expression of the PCNA in the liver of younger animals. The p53 protein was overexpressed in hyperplastic bile ducts in aged rats but not in bile ducts of younger rats (up to 24 weeks). No mutations were observed in either Ki-ras, Ha-ras, or N-ras of the liver tumors. These data suggest that point mutations in p53, Ki-ras, Ha-ras, and N-ras are not involved in the MX-induced liver carcinogenesis in rats.

K-ras and p53 mutations and overexpressions as prognostic factors in female rectal carcinoma.

BACKGROUND: The present study was undertaken to determine the prognostic significance of K-ras, p53 and bcl-2 in female rectal carcinoma. MATERIALS AND METHODS: The mutations in K-ras and p53 genes were analysed using SSCP and direct sequencing. The expression of K-ras, bcl-2 and p53 proteins was determined immunohistochemically. RESULTS: Mutations of K-ras and p53 genes were detected in 12% and 38% of the tumours, respectively. The prevalence of K-ras overexpression was 67%. K-ras mutations were not associated with survival. However, more favourable survival was observed for patients with K-ras overexpression than with normal expression (adjusted hazard ratio from Cox model 0.4, 95% CI 0.2-0.8). Mutation or overexpression of p53 were not associated with survival. CONCLUSIONS: It may be possible, that the mutations and protein overexpression of K-ras and p53 in female rectal carcinoma have different clinical impact on patient survival as suggested in previous studies concerning colorectal carcinoma of both sexes.

Physicians' gender and clinical opinions of reproductive health matters.

The purpose of this study was to investigate the effect of physicians' gender when sex-specific conditions are being treated in a country where almost half of the physicians are women. Five Finnish surveys originally conducted for other purposes were reanalyzed. Two surveys examined physicians' and medical students' views about hormone therapy during and after menopause, one survey examined medical students' attitudes about medicines, one survey examined physicians' obstetric practices, and one examined contraceptive counselling. All the surveys showed more similarities than differences in the clinical decision-making of women and men physicians. In opinions about menopause the physician's medical specialty was more important than gender. Physicians' obstetrical practices were also similar in five cases out of six, but in one situation where medical factors were not a deciding issue, women physicians acted differently. These results suggest that socialization into the medical profession makes physicians' practices more alike and diminishes gender differences.

PIP: To determine the effect of gender on the type of reproductive health treatment offered to women in a setting where half of the physicians are female, this study reanalyzed data collected from five surveys in Finland. The surveys were originally conducted to investigate the views of physicians (74% of 500 responding) and medical students (74% of 125 responding) about menopausal hormone replacement therapy, the attitudes of medical students about prescribing hormones, obstetric practices (83% of 90 responding), and contraceptive counseling (74% of 480 responding). It was found that the number of menopausal patients was highest for female gynecologists, followed by male gynecologists, female physicians, and male physicians. Female and male gynecologists offered similar recommendations for hormone replacement therapy, but female physicians were more likely to prescribe hormones for prevention only. Results for medical students were not influenced by gender. The only significant difference in obstetric practice was that female obstetricians were three times more likely than males to induce labor for convenience. While equal numbers of female and male physicians prescribed oral contraceptives and IUDs, females were more likely to prescribe condoms, implants, and diaphragms. It is concluded that professional identification among these physicians was stronger than gender influences.

Pros and cons of intrauterine contraception--do perceptions of users and physicians differ?

OBJECTIVE: To investigate women's and physicians' opinions of IUD use. DESIGN: Two cross-sectional postal surveys; on women in 1987-1988, on physicians in 1988. SETTING: Survey on women: southernmost Uusimaa province (including Helsinki). Survey on physicians: whole of Finland. PARTICIPANTS: Of a stratified random sample of 1000 women aged 18-44 years 84% returned the questionnaire (N = 844). Of a sample of 480 physicians (including GPs and gynaecologists) 418 were eligible, 74% of whom responded (N = 311). MAIN OUTCOME MEASURES: Women's and physicians' opinions of and experiences with IUD use as reported in postal questionnaires. RESULTS: Most women considered that IUDs were a good method of contraception. Users' and physicians' reports on benefits and disadvantages of IUDs metched each other; both most often mentioned ease of use, efficacy, and lack of systemic effects. Major disadvantages reported by women were bleeding, pain, infection, and pregnancy. Physicians further pointed to the risk of ectopic pregnancy. Though reports of side-effects were commonplace, there was reasonable satisfaction with IUD use. CONCLUSION: This study reveals that most women, and physicians, have a realistic picture of the common problems related to intrauterine contraception. It also emphasizes that, when making contraceptive choices, one is bound to compare possible disadvantages with benefits offered.

Problems in distribution of scientific knowledge: intrauterine contraceptive devices and drug catalogs.

Intrauterine contraceptive devices (IUDs) are a popular method of contraception worldwide. However, some serious problems have been associated with them. Finland has developed and now manufactures and exports IUDs. Therefore, drug control and the quality of drug information existing in Finland are significant for other countries, as well. This study analyzes the information in the Finnish commercial drug catalog on copper-releasing IUDs and compares it with the scientific literature, the instructions from the licensing authority, and material in its U.S. counterpart, during the last two decades. The results indicate that the distribution of scientific knowledge to the drug catalogs has often been slow. In the early 1980s Finnish manufacturers did not give any practical information on their products, and then and later the Finnish catalog was less comprehensive than the U.S. catalog. The variations in the control system in different nations were reflected in the contents of the Finnish catalog. For practitioners, drug catalogs are important sources of medical information. The results of this study demonstrate (1) that more attention should be paid to the contents of these catalogs, and (2) the continuous need for up-to-date, unbiased drug information.

PIP: The University of Helsinki's Department of Public Health has analyzed information in the Finnish Commercial drug catalog (Pharmaca Fennica [PF] on copper releasing IUDs and compared it with the scientific literature, guidelines from the licensing authority, and information in its US counterpart, the Physician's Desk Reference (PDR). The analyzed information was mostly from the 1980s. The quality and the quantity of the information varied depending on the nation publishing the catalog, and, in the Finnish catalog, the manufacturer's mother country also had an effect. Even though there was a revision and publication lag for manufacturers' drug catalogs, by 1981, the information in the US PDR was very extensive. Yet, the Finnish PF did not have information on copper releasing IUDs by 1981, even though Finnish IUDs comprised more than 80% of IUDs available on the market. In the 1980s, entries in the Finnish drug catalogs had not listed the consensus about the contraindications and side effects for IUD use found in the literature. External pressure forced the manufacturers to eventually include contraindications and side effects. Moreover, drug catalogs did not include the information recommended by the licensing authority. Studies of physician habits indicated that physicians pay considerable attention to drug advertisements and salespersons; thus, they might put healthy women at risk. Also, since communication through medical journals is faulty, pharmacopeias contain information of no practical value to physicians tend to pursue continuing medical education in areas in which they have an interest, the issue of factual information in drug catalogs is of even greater concern. Therefore, physicians and public health professionals must ceaselessly meet the need for practically relevant, unbiased, and up-to-date drug information.

Is users' knowledge about contraceptives adequate? A case study of Finnish IUD users.

Intrauterine contraceptive devices (IUDs) are popular: in Finland, in 1987, about 25% of women of childbearing age used them. We studied Finnish women's knowledge about contraindications for and side-effects of IUD use, with emphasis on current users. In December 1987-January 1988, a postal questionnaire was sent to a random sample of 1000 women of childbearing age, in Uusimaa province, which includes the capital, Helsinki. After three reminders, the response rate was 84%. We compared the answers of current and past users and nonusers of IUDs, and analyzed the associations between IUD use, education, age, and knowledge about side-effects. In contrast to the recommendations for use and information to be given to the contraceptive users, a substantial proportion of current IUD users assumed there are few limitations for IUD use. Some side-effects possibly leading to severe consequences were not considered to be connected with IUD use, not even by users. Although risk of infection was a quite well known fact, infertility was seldom associated with IUD use, and 22% of current IUD users did not associate ectopic pregnancy with IUDs. The results thus suggest that contraceptive counseling is not fully adequate and should receive more attention.

DNA profiling in a genetically isolated population using three hypervariable DNA markers.

We determined the allele frequencies for three hypervariable DNA loci D2S44, D1S7 and D7S21 using the probes YNH24, MS1 and MS31 in the genetically isolated Finnish population. The allelic length ranges were 1.7- < 6.0 kb for YNH24, 1.7- < 18.0 kb for MS1 and 3.2- < 12.0 kb for MS31. High heterozygosity rates (0.94-0.96) were detected for all three probes. In 48 mother-child pairs no mutations were found using the probes YNH24 and MS31, whereas a mutation rate of 0.064 was observed for probe MS1. In addition, an unexpected four-band pattern was detected in 1 out of 170 individuals using the probe MS1, suggesting complex DNA polymorphism based on both a variable number of tandem repeats and restriction site polymorphism. Our findings also show that all three probes are valuable in individual identification in this genetically isolated population.

Different contraceptive practices: use of contraceptives in Finland and other Nordic countries in the 1970s and 1980s.

Contraceptive practices, especially oral contraceptive and intrauterine device use, were studied in four Nordic countries by recalculating published and unpublished data from previous surveys and statistics and by collecting new data from Finland. The sales of oral contraceptives were presented in defined daily doses, and the percentages of women using oral contraceptives were estimated from them. The percentages of intrauterine device users were calculated taking into account the number of intrauterine devices sold each year and the continuation of use from previous years. The results of the surveys were reanalysed. We found clear differences in contraceptive practices. Oral contraceptive use was most prevalent in Sweden and Denmark, and, especially at the end of the 1970s, Finnish intrauterine device use was very high. In the 1980s the differences dimished somewhat. These disparities in culturally and economically similar countries indicate that further research is needed to evaluate the factors influencing contraceptive practices.

PIP: Researchers used sales data and data from fertility surveys to compare contraceptive practices among the 4 Scandinavian countries. Sales of oral contraceptives (OCs) remained relatively stable within each country, but women used OCs more often in Sweden and Denmark than in Finland and Norway. By the end of the 1980s, however, OC use was about the same in all 4 countries. IUD sales increased considerably more in the late 1970s in Finland than Denmark and Norway. The researchers could not find reliable IUD sales data for Sweden after 1981. IUD sales decreased in the late 1980s in all 3 countries, however. These figures did not take into account the fact that the life time of an IUD increased at this time. In addition, sales figures may have overestimated IUD users by 5-10%. In 1971, the condom ranked as the most prevalent contraceptive used among Finnish women (27%) followed by OCs (17%) and withdrawal (15%). In 1981, most Finnish women used the IUD (25%), especially copper releasing IUDs, followed by the condom (20%) and OCs (18%). In 1977, women tended to use the IUD (23%) in Norway. In 1981, the most popular family planning methods in Sweden were barrier methods (21%), OCs (19%), and the IUD (17%). In 1983, 24% of women in Denmark used OCs. An overall view showed that women in Finland and Sweden preferred to use OCs and IUDs than did women in Norway and Denmark. Maternity clinics in Finland tended to favor IUDs when offering contraceptive advise. In conclusion, researchers should conduct studies to determine those factors which influence contraceptive practices.