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Complex formation and liquid-liquid extraction in a system containing iron(III), 4-nitrocatechol (4NC), 2,3,5-triphenyl-2H-tetrazolium chloride (TTC), water, and chloroform were studied. The effect of some experimental parameters (pH, shaking time, concentration of reagents) was investigated, and the optimum conditions for Fe(III) extraction as an ion-association complex, (TT+)3[Fe3+(4NC)3]3-, were found. The following key constants were calculated: constant of distribution (Log KD=1.49±0.02), constant of association (Log β=11.3±0.1), and constant of extraction (Log Kex=12.8±0.1). Some additional characteristics concerning the application of 4NC and TTC for extractive-spectrophotometric determination of Fe(III) were estimated as well: absorption maximum (λ=435 nm), apparent molar absorptivity (ε=2.7×104 l mol-1 cm-1), Sandell's sensitivity (SS=2.0 ng cm-2), recovery factor (R %=94.3±0.2), limit of detection (LOD=0.11 μg cm-3), and limit of quantification (LOQ=0.37 mg cm-3). Beer's law was obeyed for Fe(III) concentrations up to 2.0 μg ml-1 with a correlation coefficient of 0.9990. [ABSTRACT FROM AUTHOR]

Objectives: To investigate whether treatment effects of pharmaceutical compounds compared with placebo controls are systematically different to the effects of the same compounds compared with active treatment controls in rheumatoid arthritis (RA) clinical trials., Methods: We systematically identified randomised controlled trials (RCTs) in RA, and matched active treatment arms with comparable regimens, populations, background therapy, and outcome reporting, by the nature of their control group (active comparator or placebo). Medline, EMBASE and CENTRAL were used to identify RCTs investigating disease modifying anti-rheumatic drug therapies until December 2021. Using mixed-model logistic regression we estimated OddsRatios (OR) for achieving an American College of Rheumatology (ACR) 20/50/70% response at weeks 12 and 24. Risk of bias was assessed using the Cochrane Tool., Results: We screened 8328 studies and included 40 for analysis after detailed review of 590 manuscripts; unique compounds had significantly higher responses in active comparator trials compared with their effects observed in placebo controlled trials, with ORs of 1.67 (95% CI 1.46 to 1.91; p<0.001) for ACR20, 1.50 (95% CI 1.29 to 1.75; p<0.001) for ACR50 and 1.65 (95% CI 1.30 to 2.10; p<0.001) for ACR70 (week 12); corresponding ORs for ACR 20, 50, and 70 (week 24) were 1.93 (95% CI 1.50 to 2.48; p<0.001), 1.75 (95% CI 1.32 to 2.33; p<0.001) and 1.68 (95% CI 1.21 to 2.34; p<0.001), respectively. Sensitivity analyses showed consistent results., Conclusion: Placebo controlled trials lead to smaller effect sizes of active compounds in RCTs compared with the same compound in head-to-head trials. This difference may be explained by potential nocebo effects in placebo-controlled settings and needs to be considered when interpreting head-to-head and placebo-controlled trials, by patients, investigators, sponsors and regulatory agencies., Competing Interests: Competing interests: AK: Speakers bureau, Consultancy: AbbVie, Amgen, Bristol-Myers Squibb, Eli Lilly, Gilead, Janssen, Merck Sharp and Dohme, Novartis, UCB and Pfizer. NMS: Nothing to declare. TS: Nothing to declare. EC: Nothing to declare. JSS: received grants from Abbvie, Astra-Zeneca, Janssen, Lilly, Novartis, Roche and honoraria from Abbvie, Amgen, Astra-Zeneca, Astro, BMS, Celgene, Celltrion, Chugai, Gilead, ILTOO, Janssen, Lilly, MSD, Novartis-Sandoz, Pfizer, Roche, Samsung, Sanofi, UCB. DA received grants from Abbvie, Amgen, Lilly, Novartis, Roche, SoBi, Sanofi; and honoraria (speakers bureau/consultancy) from Abbvie, Amgen, Lilly, Janssen, Merck, Novartis, Pfizer, Roche, Sandoz., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Objectives: The purpose of this study was to identify lifestyle patterns that are characteristic of overweight and obese European adults in the context of educational level., Methods: This was a cross-sectional study of dietary data from 1235 men and 10 343 women. Dietary intake, educational level, and physical activity were assessed using questionnaires. A principal component analysis was used to derive lifestyle patterns, and associations with being overweight or obese (OW/OB) and waist circumference (WC) were explored by applying a multivariate logistic regression., Results: Overall, 35% of women and 68% of men were OW/OB, of whom 30% and 40%, respectively, had ≤12 y of education. The principal component analysis derived 2 distinct dietary patterns (healthy vs. unhealthy). The daily intake of fruits (fresh, canned, and juice) and vegetables was found to be associated with lower odds of being OW/OB and WC in women only. In contrast, the daily intake of diet soft drinks was associated with higher odds of being OW/OB in women and men, but the daily intake of sweets was associated with higher odds of WC in women only. In both sexes, having &gt;12 y of education was inversely associated with being OW/OB. No associations were observed for regular soft-drink intake., Conclusions: A healthy dietary pattern constituting of a daily intake of fruits and vegetables was inversely associated with being OW/OB in women. Conversely, a diet soft-drink intake was positively associated with being OW/OB in both sexes, probably because of a reduction in overall energy intake. More studies are recommended to clarify the effectiveness of diet soft-drink consumption in controlling caloric intake and as a healthier alternative to regular soft drinks and sweets., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The Feel4Diabetes project conducted a 2-year intervention aiming to encourage healthy behaviours in families in six European countries. Within this paper, the intervention effect was evaluated after the first intervention year on 24-hour physical behaviour composition in Belgian families at risk for type 2 diabetes. Parents' and children's levels of physical activity (PA) and sedentary behaviour (SB) were objectively measured and sleep duration was self-reported. Valid data were obtained in 82 children (mean age: 8.2 ± 0.84; 50.0% boys) and 72 parents (mean age: 40.1 ± 5.5; 27.8% fathers). Data were analysed using a compositional data approach, and mixed models were used to take clustering into account. Results showed that for children, the mean baseline composition of the 24-hour day in sleep, SB, light and moderate-to-vigorous PA for the intervention group was (11 hours 18 minutes; 6 hours 33 minutes; 5 hours 02 minutes, 1 hour 08 minutes) and for the control group (11 hours 18 minutes; 6 hours 35 minutes; 5 hours 04 minutes; 1 hour 06 minutes), respectively. For parents, the mean baseline composition was for the intervention group (8 hours 12 minutes; 9 hours 36 minutes; 5 hours 43 minutes; 27 minutes) and for the control group (8 hours 00 minute; 9 hours 00 minute; 6 hours 27 minutes; 33 minutes). No significant intervention effects were found on 24-hour physical behaviour composition of either parents or children (p = 0.19 and p = 0.21, respectively). A relatively small study population and a poor attendance rate among parents could maybe explain the lack of effectiveness. More effective strategies are needed to retain vulnerable families in interventions. Furthermore, future approaches to improve the 24-hour physical behaviour composition should maybe focus more specifically on PA, SB and sleep, and involve teachers, children and parents in the design of the intervention. Trial registration: NCT02393872 in ClinicalTrials.gov., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The Feel4Diabetes study recruited 12,193 children (age: 8.20 ±1.01 years) and their parents from six European countries as part of the broader attempt to prevent type 2 diabetes. The current work collected data pre-intervention to identify the prevalence of childhood obesity by country and describe its association with socio-demographic characteristics and parental obesity status. One in four children were overweight or obese, and one in four families had at least one obese parent. Multivariate logistic regression examined the associations between childhood obesity, family socio-demographics, and parental obesity status. Children had a higher chance of being overweight or obese if they were living in “low income” countries (OR: 2.11, 95% CI: 1.62, 2.74) and countries “under economic crisis” (OR: 2.48, 95% CI: 1.89, 3.24) compared to “high-income” countries; if their fathers completed fewer than nine years of education (OR: 2.16, 95% CI: 1.54, 3.05) compared to children whose fathers had a higher level (>14 years) of education; and if one (OR: 2.46, 95% CI: 0.32, 0.62) or both of their parents (OR: 6.83, 95% CI: 5.15, 9.05) were obese. Future childhood obesity prevention-programs should target the whole family while taking into consideration the socioeconomic and weight status of parents. Future research should examine these associations in more countries and in socio-demographically diverse populations in order to facilitate the generalisability of the present study’s findings.

Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only live attenuated vaccine available against tuberculosis. The first BCG vaccination was done exactly 100 years ago, in 1921. The BCG vaccine strains used worldwide represent a family of daughter sub-strains with distinct genotypic characteristics. BCG SL222 Sofia is a seed lot sub-strain descending from the Russian BCG-I (seed lot 374a) strain and has been used for vaccine production in Bulgaria since 1972. Here, we report the assembled circular genome sequence of Mycobacterium bovis BCG SL222 Sofia and phylogeny analysis with the most closely related BCG sub-strains. The full circular genome of BCG SL222 Sofia had a length of 4,370,706 bp with an average GC content of 65.60%. After 49 years of in vitro evolution in a freeze-dried condition, we identified four SNP mutations as compared to the reference BCG-I (Russia-368) sequence. BCG vaccination is of central importance for the TB elimination programs in many countries. Since 1991, almost 40 million vaccine doses of the BCG SL222 Sofia have been distributed annually through the United Nations Children's Fund (UNICEF) and the Pan American Health Organization (PAHO) to approximately 120 countries. The availability of the complete reference genome sequence for M. bovis BCG SL222 Sofia, a WHO reference reagent for the Russian BCG-I sub-strain, will facilitate the identity assurance of the genomic stability, will contribute to more consistent manufacturing, and has an important value in standardization and differentiation of sub-strains used in vaccine production. We propose to rename the sub-strain BCG SL222 Sofia to BCG-Sofia for practical and common use.

Mycobacterium bovis BCG SL222 Sofia is a substrain descending from the Russian BCG-I vaccine strain. Here, we report the complete genome sequence of BCG SL222 Sofia, which will facilitate identity assurance and will contribute to more consistent manufacturing, standardization, and differentiation of substrains used in vaccine production., (Copyright © 2021 Panaiotov et al.)

Phase 3 trials are the mainstay of drug development across medicine but have often not met expectations set by preceding phase 2 studies. A systematic meta-analysis evaluated all randomized controlled, double-blind trials investigating targeted disease-modifying anti-rheumatic drugs in rheumatoid and psoriatic arthritis. Primary outcomes of American College of Rheumatology (ACR) 20 responses were compared by mixed-model logistic regression, including exploration of potential determinants of efficacy overestimation. In rheumatoid arthritis, phase 2 trial outcomes systematically overestimated subsequent phase 3 results (odds ratio comparing ACR20 in phase 2 versus phase 3: 1.39, 95% confidence interval: 1.25-1.57, P < 0.001). Data for psoriatic arthritis trials were similar, but not statistically significant (odds ratio comparing ACR20 in phase 2 versus phase 3: 1.35, 95% confidence interval: 0.94-1.94, P = 0.09). Differences in inclusion criteria largely explained the observed differences in efficacy findings. Our findings have implications for all stakeholders in new therapeutic development and testing, as well as potential ethical implications.

The purpose of the present study was to analyze the concentrations of lead, cadmium and mercury in mussels (Mytilus galloprovincialis) and veined rapa whelks (Rapana venosa) caught in the Varna Bay of Black Sea and to evaluate the risk for human health from the presence of the three heavy metals. The highest average concentrations in mussels were those of cadmium (0.280 mg/kg), followed by lead (0.251 mg/kg) and mercury (0.017 mg/kg). Veined rapa whelks also showed highest levels of cadmium (1.113 mg/kg), followed by lead (0.045 mg/kg) and mercury (0.034 mg/kg). EDI values for adults consuming mussels and veined rapa whelks were below the published RfDo and PTWI values. All THQ and HI values were below 1. The consumption of M. galloprovincialis and R. venosa caught in the Varna Bay, Black Sea, did not pose any risk for the health of adult people as lead, cadmium and mercury were concerned., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Antibody repertoires of healthy humans and animals contain a fraction of antibodies able to acquire additional polyspecificity following exposure to several biologically relevant redox molecules (free heme, reactive oxygen species, ferrous ions, HOCl, etc.). The physiological role of these "hidden" polyspecific antibodies is poorly understood. Similar to inherently polyspecific antibodies, those with induced polyspecificicty may also have immunoregulatory properties. We have previously shown that a pooled human IgG preparation, modified by the exposure to ferrous ions, acquires the ability to significantly improve survival of animals with polymicrobial sepsis or aseptic systemic inflammation induced by bacterial lipopolysaccharide or zymosan administration. In the present study, we have analyzed the effects of administration of heme-exposed pooled human IgG in the same models of sepsis and aseptic systemic inflammation. The administration of a single dose of heme-exposed pooled IgG has resulted in a significant increase in the survival of mice with endotoxinemia, but not in those with polymicrobial sepsis and zymosan-induced severe generalized inflammation. Finally, we have provided evidence that the anti-inflammatory effect of heme-exposed IgG can be explained by scavenging of pro-inflammatory mediators.

Crashes at level crossings are a major issue worldwide. In Australia, as well as in other countries, the number of crashes with vehicles has declined in the past years, while the number of crashes involving pedestrians seems to have remained unchanged. A systematic review of research related to pedestrian behaviour highlighted a number of important scientific gaps in current knowledge. The complexity of such intersections imposes particular constraints to the understanding of pedestrians' crossing behaviour. A new systems-based framework, called Pedestrian Unsafe Level Crossing framework (PULC) was developed. The PULC organises contributing factors to crossing behaviour on different system levels as per the hierarchical classification of Jens Rasmussen's Framework for Risk Management. In addition, the framework adapts James Reason's classification to distinguish between different types of unsafe behaviour. The framework was developed as a tool for collection of generalizable data that could be used to predict current or future system failures or to identify aspects of the system that require further safety improvement. To give it an initial support, the PULC was applied to the analysis of qualitative data from focus groups discussions. A total number of 12 pedestrians who regularly crossed the same level crossing were asked about their daily experience and their observations of others' behaviour which allowed the extraction and classification of factors associated with errors and violations. Two case studies using Rasmussen's AcciMap technique are presented as an example of potential application of the framework. A discussion on the identified multiple risk contributing factors and their interactions is provided, in light of the benefits of applying a systems approach to the understanding of the origins of individual's behaviour. Potential actions towards safety improvement are discussed., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Despite the intensive research in the past decade on the microbial bioaccumulation of heavy metals, the significance of redox state for oxidative stress induction is not completely clarified. In the present study, we examined the effect of redox-active (copper and chromium) and redox-inactive (cadmium) metals on the changes in levels of oxidative stress biomarkers and antioxidant enzyme defence in Trichosporon cutaneum R57 cells. This filamentous yeast strain showed significant tolerance and bioaccumulation capability of heavy metals. Our findings indicated that the treatment by both redox-active and redox-inactive heavy metal induced oxidative stress events. Enhanced concentrations of Cu 2+ , Cr 6+ and Cd 2+ caused acceleration in the production of reactive oxygen species (ROS), increase in the level of oxidatively damaged proteins and accumulation of reserve carbohydrates (glycogen and trehalose). Cell response against heavy metal exposure also includes elevation in the activities of antioxidant enzymes, superoxide dismutase and catalase, which are key enzymes for directly scavenging of ROS. Despite the mentioned changes in the stress biomarkers, T. cutaneum did not show a significant growth diminution. Probably, activated antioxidant defence contributes to the yeast survival under conditions of heavy metal stress.

Tyrphostin AG490 is a Janus kinase (JAK) 2 inhibitor that is clinically used as an anticancer agent and is also effective in various models of inflammatory and autoimmune diseases. In this study, we examined the effects of tyrphostin AG490 on the development of collagenase-induced osteoarthritis (CIOA). Our results showed that tyrphostin-ameliorated cartilage and bone destructions. This effect was associated with decreased expression of signal transducers and activators of transcription 3 (STAT3), phosphorylated JAK2, Dickkopf homolog 1, and receptor activator of nuclear factor κB ligand (RANKL) in the joints of arthritic mice. Tyrphostin AG490 suppressed STAT3 phosphorylation and the expression of tumor necrosis factor-related apoptosis-inducing ligand and RANKL by synovial fluid cells. The drug inhibited RANKL-induced osteoclast differentiation in vitro. Molecules, such as tyrphostin AG490 that limit bone erosion and influence osteoclast generation, might have therapeutic utility in joint degenerative disorders.

Coumarin and its derivatives have potent immunomodulatory activities. Here we describe the parameters of the protective effect of 7-hydroxycoumarin (7-OHC) in experimental Salmonella enterica Serovar Typhimurium infection in mice. The protective effect depended on the duration of treatment reaching its maximum after 10 days of pretreatment and lasted for at least 15 days after its end. Electron microscopy studies revealed that 7-OHC induced ultrastructural changes in macrophages consistent with their activation as well as faster destruction of ingested salmonellae. Superoxide and hydrogen peroxide secretion by macrophages was decreased in both healthy and Salmonella-infected 7-OHC treated animals, which is in line with the current view that some coumarins possess antioxidant and radical scavenging activity. Thus, 7-OHC pretreatment also appears beneficial to the host by limiting the harmful tissue damaging and immunosuppressive effects of the oxidative stress during a Salmonella infection but still activates the microbicidal capacity of exposed phagocytes.

The combined protective effect of a polyphenol-rich extract, isolated from Geranium sanguineum L. (PC), and a novel naturally glycosylated Cu/Zn-containing superoxide dismutase, produced from the fungal strain Humicula lutea 103 (HL-SOD), in the experimental influenza A virus infection (EIVI) in mice, induced with the virus A/Aichi/2/68 (H3N2), was investigated. The combined application of HL-SOD and PC in doses, which by themselves do not defend significantly mice in EIVI, resulted in a synergistically increased protection, determined on the basis of protective indices and amelioration of lung injury. Lung weights and consolidation as well as infectious lung virus titers were all decreased significantly parallel to the reduction of the mortality rates; lung indices were raised. The excessive production of reactive oxygen species (ROS) by alveolar macrophages (aMphi) as well as the elevated levels of the lung antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), induced by EIVI, were brought to normal. For comparative reasons the combined protective effect of PC and vitamin C was investigated. The obtained results support the combined use of antioxidants for the treatment of influenza virus infection and in general indicate the beneficial protective role of combinations of viral inhibitors of natural origin with diverse modes of action.

New La(III) and Dy(III) complexes of deprotonated 4-hydroxy-3[1-(4-nitrophenyl)-3-oxobutyl]-2H-1-benzopyran-2-one (Acenocoumarol) were synthesized and characterized using FT-IR, FT-Raman, (1)H NMR spectra, and elemental analyses. The ligand and its lanthanide(III) complexes were tested for their cytotoxic/cytostatic activity against two tumor cell lines and peritoneal mouse macrophages. The La(III) and Dy(III) complexes exhibit good activity against melanoma B16 and fibrosarcoma L929 and they are stronger inhibitors of tumor cell proliferation compared to the ligand without influencing normal cell viability and NO release by mouse peritoneal macrophages., (Copyright 2009 Elsevier GmbH. All rights reserved.)

A new hemocyanin was isolated from the hemolymph of garden snails Helix vulgaris, composed of two isoforms, HvH1 and HvH2 separated on an ion exchange column DEAE-Sepharose 6CL. Structural and immunological properties of Helix vulgaris hemocyanin were studied in comparison with molluscan Hcs Rapana venosa and Megathura crenulata. The possibility of using HvH and RvH as carriers of small molecules (haptens) in immunizing protocols was studied in comparison with KLH, which is a widely used, highly immunogenic carrier protein. By using HvH as a carrier of the well-known hapten TNBS (2,4,6-trinitrobenzene sulfonic acid), an increasing with time production of hapten-specific TFN-gamma was detected in splenocyte cultures of mice, which lasted longer than in case of KLH and RvH carriers. Also, use of HvH or RvH as a carrier of the hapten ProT alpha[101-109] (i.e., the synthetic C-terminal fragment of the poorly immunogenic protein prothymosin alpha) showed that antisera of higher titres than that of the control conjugate (ProT alpha[101-109]-KLH) were obtained immediately after the second bleeding. HvH and RvH may prove to be useful for the development of new antiviral, antibacterial and antitumor vaccines, since they seem to launch strong and specific immune response against the conjugated antigens.

Coumarin and its derivatives are naturally occurring substances with multiple biological activities. Here we demonstrate that prophylactic peroral administration of coumarin or 7-hydroxycoumarin (7-OHC) enhances resistance to subsequent lethal Salmonella enterica Serovar Typhimurium infection in mice. 7-OHC decreased bacterial load in liver and spleen, and enhanced phagocytosis and bacterial killing by macrophages when applied in vitro and in vivo. 7-OHC treatment induced significant NO release in peritoneal macrophage cultures. The observed protective effect correlated with the induction of Th1-associated cytokines, such as IL-12, IFN-gamma, and TNF-alpha. These data demonstrate a clear immunomodulatory potential of coumarins which might have important therapeutic implications to enhance resistance to infection.

Influenza infection was induced in white mice by intranasal inoculation of the virus A/Aichi/2/68 (H3N2). The lung protease and the protease-inhibitory activities were followed for 9 days after infection. The intranasal application of a polyphenol-rich extract (PC) isolated from Geranium sanguineum L. induced a continuous rise in the anti-protease activity but did not cause substantial changes in the lung protease activity of healthy mice. Influenza virus infection triggered a slight reduction in protease activity in the lungs at 5 and 48 h post infection (p.i.) and a marked increase at 24 h and 6 day p.i.. Protease inhibition in the lungs was reduced at 24 and 48 h p.i. and an increase was observed at 5 h and 6 and 9 days p.i.. PC treatment brought both activities to normal levels. The restoration of the examined parameters was consistent with a prolongation of mean survival time and reduction of mortality rate, infectious virus titre and lung consolidation. PC reinstated superoxide production by alveolar macrophages and increased their number in virus-infected mice. The favourable effect on the protease and the protease-inhibitory activities in the lungs of influenza-virus-infected mice apparently contributes to the overall protective effect of PC in the murine experimental influenza A/Aichi infection. The antiviral effect of the individual constituents was evaluated.

A small cryptic Lactobacillus helveticus plasmid, pLBL4, was able to reveal restriction fragment length polymorphism in different bacterial species including Lactobacillus species, Bacillus species, and Escherichia coli when used as a DNA probe. The observed polymorphism was a result of the combined hybridization of several microsatellite sequences. The 6-bp sequence (TTGTTT) was repeated 12 times, seven of which were concentrated within the region between 1791 and 1997 bp of the plasmid sequence. The polymorphic patterns generated with pLBL4 differed from those obtained with M13 DNA in the larger number of bands observed. The results presented here open the possibility of using pLBL4 as a new broad-spectrum polymorphic DNA probe for fingerprint analysis.

A result of extensive screening of over 300 strains from the Collection of ELBY Bulgaricum, PLC, thirty six strains were selected as producers of bacteriocins, active closely related lactic acid bacterial species and some food spoilage bacteria. The selected strains belong to L. helveticus, L. bulgaricus and S. thermophilus, which are rare bacteriocin producers. Nineteen nonidentified producers were characterized by molecular taxonomic approaches--M13 fingerprinting, repetitive PCR, ribotyping and hybridization with species-specific probes, which allowed to affiliate them to the species L. delbrueckii. Several strains were found to harbour plasmids of different size. The estimated activity against food borne pathogens makes the isolated substances perspective as safe food preservatives and the producing strains could be used as components of starters with improved quality.

Three different cryptic plasmids from Lactobacillus helveticus have been identified and their DNA sequences determined. Analysis and comparisons of their primary structures revealed stretches of DNA with considerable homology. Thus, large portions of the plasmid non-coding sequences were conserved at 80-90% identity between the different plasmids identified so far in L. helveticus. Nevertheless, different plasmids found in a same host strain utilise different genes of replication, probably acquired during evolution from different replicons from Gram-positive bacterial origins. A remnant structure of such a possible genetic integration of a foreign replication gene into one of the plasmids of L. helveticus was identified.

The optimal conditions for the application of M13 DNA fingerprinting to the genus Lactobacillus were determined. Comparative fingerprint analysis of representative strains of Lactobacillus delbrueckii subsp. delbrueckii, Lact. delbrueckii subsp. lactis, Lact. delbrueckii subsp. bulgaricus, Lact. helveticus and Lact. casei permitted the differentiation of species, subspecies and individual strains and the quantitative determination of their genetic relatedness. The results confirm the high specificity of M13 DNA fingerprinting and indicate that it might be used in the classification of Lactobacillus spp.

The authors describe a method for determination of estrogens in urine of nonpregnant women and men. The method includes enzymic hydrolysis of urine, gel filtration of Sefadex G25, extraction with organic solvents and modifies Kober reaction with spectrofluorometric determination. The technique of the method is not complex, the usage of reagents is small and is suitable for everyday work.