Assessing testicular germ cell DNA damage in the comet assay; introduction of a proof-of-concept.

The in vivo comet assay is widely used to measure genotoxicity; however, the current OECD test guideline (TG 489) does not recommend using the assay to assess testicular germ cells, due to the presence of testicular somatic cells. An adapted approach to specifically assess testicular germ cells within the comet assay is certainly warranted, considering regulatory needs for germ cell-specific genotoxicity data in relation to the increasing global production of and exposure to potentially hazardous chemicals. Here, we provide a proof-of-concept to selectively analyze round spermatids and primary spermatocytes, distinguishing them from other cells of the testicle. Utilizing the comet assay recordings of DNA content (total fluorescence intensity) and DNA damage (% tail intensity) of individual comets, we developed a framework to distinguish testicular cell populations based on differences in DNA content/ploidy and appearance. Haploid round spermatid comets are identified through (1) visual inspection of DNA content distributions, (2) setting DNA content thresholds, and (3) modeling DNA content distributions using a normal mixture distribution function. We also describe an approach to distinguish primary spermatocytes during comet scoring, based on their high DNA content and large physical size. Our concept allows both somatic and germ cells to be analyzed in the same animal, adding a versatile, sensitive, rapid, and resource-efficient assay to the limited genotoxicity assessment toolbox for germ cells. An adaptation of TG 489 facilitates accumulation of valuable information regarding distribution of substances to germ cells and their potential for inducing germ cell gene mutations and structural chromosomal aberrations.

Double-balloon catheter for induction of labor in 362 women with and without prior cesarean section.

OBJECTIVE: Balloon catheter is the preferred method for induction of labor in women with prior cesarean section. We sought to evaluate the rate of vaginal delivery, induction-delivery time and outcome predictors after induction with double-balloon catheter. STUDY DESIGN: We conducted a retrospective cohort study including women with prior cesarean section undergoing induction of labor with a double-balloon catheter during the period January 2007-June 2014 at a large, tertiary Danish university hospital. For comparison, we included women with no prior cesarean section undergoing induction with double-balloon catheter after failed medical induction. Inclusion criteria were singleton pregnancy, an unfavorable cervix, intact membranes, cephalic presentation and either previous cesarean section or failed medical induction of labor. Exclusion criteria included contraindications for vaginal delivery, severe fetal malformation and stillbirth. Study subjects were identified in a local computerized system and data extracted from the medical records. RESULTS: Women with prior cesarean section (n = 304) induced with double-balloon catheter had a vaginal delivery rate of 50.3% (95% CI 44.7-55.9) compared to 51.7% (95% CI 39.2-64.1) in women with no prior cesarean section but preceding failed medical induction of labor (n = 58) (p = 0.85). BMI≥30 was associated with increased frequency of cesarean section. Median time from induction to vaginal delivery was 27.1(20.4-31.1) hours and 28.4(25.5-36.1) hours, respectively (p = 0.05). The rate of complete uterine rupture was 1.0%. CONCLUSIONS: Similar success rates of approximately 50% for vaginal delivery were observed after induction of labor with a double-balloon catheter in women with and without prior caesarean section. A BMI ≥ 30 was associated with an increased frequency of caesarean section.

Fatty Liver Among Adolescent Offspring of Women With Type 1 Diabetes (the EPICOM Study).

OBJECTIVE: Intrauterine exposure to maternal type 1 diabetes is associated with a less favorable metabolic profile later in life. Nonalcoholic fatty liver disease is the hepatic manifestation of a cluster of metabolic abnormalities linked to insulin resistance. This study aimed to evaluate the effect of maternal pregestational type 1 diabetes on the presence of fatty liver in offspring and the association between maternal BMI, glycemic control during pregnancy, offspring metabolic risk factors, and offspring level of soluble CD163 (sCD163) (a marker of macrophage activation) and risk of fatty liver. RESEARCH DESIGN AND METHODS: This study was a prospective nationwide follow-up study of offspring (n = 278) of mothers with pregestational type 1 diabetes between 1993 and 1999 and matched control subjects (n = 303). Mean age at the time of follow-up was 16.7 years (range 13.0-20.4 years). We used the fatty liver index (FLI) and waist-to-height ratio (WHtR) to evaluate the presence of fatty liver among the offspring. An FLI ≥60 or WHtR >0.469 were used as cutoff points for fatty liver. RESULTS: More type 1 diabetes-exposed offspring had high FLI and WHtR indices compared with unexposed control subjects. We found significant associations between increasing maternal prepregnancy BMI, being born large for gestational age, offspring level of sCD163, as well as offspring metabolic risk factors (decreasing adiponectin and HDL cholesterol and increasing leptin, HOMA of insulin resistance, and HOMA of insulin secretion) and degree of fatty liver. CONCLUSIONS: Intrauterine exposure to maternal type 1 diabetes and higher maternal prepregnancy BMI may predispose to fatty liver in the offspring. Offspring metabolic risk factors, including sCD163 levels, are associated with indices of fatty liver.

[Genital actinomycosis and pelvic abscesses in a woman with a 13-year-old intrauterine device].

A 55-year-old woman who had had the same intrauterine device (IUD) for 13 years was referred to the gynaecology outpatient clinic due to constitutional symptoms, abdominal pain and vaginal discharge. Diagnostic imaging showed multiple pelvic abscesses, and severe chronic endometritis with Actinomyces was found in an endometrial biopsy. The patient underwent surgical drainage of the accessible abscesses and started long-term antibiotic treatment. This case report illustrates that actinomycosis is an important differential diagnosis in symptomatic women with IUD and suspected gynaecologic malignancy.

[Multidisciplinary teams in cancer care].

Multidisciplinary teams (MDTs) are considered a core component of cancer care worldwide. It is commonly believed that MDTs ensure higher-quality decision-making and improved outcomes for the patients. However, the evidence underpinning MDTs is weak and the degree to which they have been absorbed into clinical practice varies widely. Based on a literature review this article explores effects, barriers and potentials for improving MDTs in cancer care. A stronger focus is needed on how MDTs can be optimized to achieve the best possible results in cancer care.

Hacking the hospital environment: young adults designing youth-friendly hospital rooms together with young people with cancer experiences.

BACKGROUND: There is a need for youth-friendly hospital environments as the ward environment may affect both patient satisfaction and health outcomes. OBJECTIVE: To involve young people in designing youth-friendly ward environment. METHODS: We arranged a design competition lasting 42 h (Hackathon). Students in architecture, design, engineering, communication and anthropology participated (27 young adults) - forming eight groups. Adolescents and young adults (AYA) with current or former cancer experience participated as sparring partners. We provided workspace and food during the weekend. The groups presented their products to a jury and relevant stakeholders. RESULTS: The groups created eight unique design concepts. The young designers were extremely flexible listening to ideas and experiences from the young patients, which led to common features including individual and flexible design, privacy in two-bed wardrooms and social contact with other hospitalized AYA. The winning project included an integrated concept for both wardrooms and the AYA day room, including logos and names for the rooms and an 'energy wall' in the day room. CONCLUSION: A hackathon event was an effective mode of youth participation. The design concepts and ideas were in line with current evidence regarding pleasing hospital environment and youth-friendly inpatient facilities and may be applicable to other young patients.

Relationship between death and infections among patients hospitalized in internal medicine departments: a prevalence and validation study.

BACKGROUND: We assessed the burden of infection-related death at internal medicine departments in Denmark and the validity of 2 population-based registries for diagnoses of infection. METHODS: We reviewed medical records of all inpatients who died at a large university hospital during 2008 with an infection diagnosis in the Cause of Death Registry (CDR) or Hospital Discharge Registry (HDR). We computed the positive predictive value of infection diagnoses and completeness of each registry with 95% confidence intervals (CIs) and the prevalence of infection-related deaths by capture-recapture analysis. RESULTS: Among 458 deaths, 193 patients (42.1%) had an infection diagnosis of which 40.0% (183 out of 458) were verified. The positive predictive value of an infection diagnosis was 96.0% (95% CI, 92.0%-98.3%) in the CDR and 95.3% (CI, 90.9%-97.9%) in the HDR. Completeness of the CDR was 79.2% (CI, 72.9%-84.6%) and completeness of the HDR was 77.0% (CI, 70.6%-82.7%). By capture-recapture analysis we estimated that 43.2% of all deaths (198 out of 458) were related to infection. CONCLUSIONS: The large proportion of deaths in internal medicine departments that are related to infection emphasizes the need for optimized infection prevention and treatment strategies. CDR and HDR are valid sources for identifying infection and may supplement each other for increased completeness of infection-related death in epidemiologic research.

In utero exposure to nanosized carbon black (Printex90) does not induce tandem repeat mutations in female murine germ cells.

Inhalation of particles has been shown to induce mutations in the male germline in mice following both prenatal and adult exposures in several experiments. In contrast, the effects of particles on female germ cell mutagenesis are not well established. Germline mutations are induced during active cell division, which occurs during fetal development in females. We investigated the effects of prenatal exposure to carbon black nanoparticles (CB) on induction of mutations in the female mouse germline during fetal development, spanning the critical developmental stages of oogenesis. Pregnant C57BL/6J mice were exposed four times during gestation by intratracheal instillation of 67µg/animal of nanosized carbon black Printex90 or vehicle (gestation days 7, 10, 15 and 18). Female offspring were raised to maturity and mated with unexposed CBA males. Expanded simple tandem repeat (ESTR) germline mutation rates in the resulting F2 generation were determined from full pedigrees (mother, father, offspring) of F1 female mice (178 CB-exposed and 258 control F2 offspring). ESTR mutation rates in CB-exposed F2 female offspring were not statistically different from those of F2 female control offspring.

Daily sperm production: application in studies of prenatal exposure to nanoparticles in mice.

We investigated the influence of maternal airway exposure to nanoparticulate titanium dioxide (TiO2, UV-Titan) and carbon black (CB, Printex90), on male reproductive function in the two following generations. Time-mated C57BL/6J mice were exposed by inhalation to UV-Titan, or by intratracheal instillation with Printex90. Body and testicle weight, sperm content per g testicular parenchyma and daily sperm production (DSP) were assessed. The protocol for assessment of DSP was optimized for application in mice (C57BL/6J) and the influence of different parameters was studied. Maternal particulate exposure did not affect DSP statistically significantly in the F1 generation, although TiO2 tended to reduce sperm counts. Overall, time-to-first F2 litter increased with decreasing sperm production. There was no effect on sperm production in the F2 generation originating after TiO2 exposure. F2 offspring, whose fathers were prenatally exposed to Printex90, showed lowered sperm production. Furthermore, we report statistically significant differences in sperm production between mouse strains.

NanoTIO(2) (UV-Titan) does not induce ESTR mutations in the germline of prenatally exposed female mice.

BACKGROUND: Particulate air pollution has been linked to an increased risk of cardiovascular disease and cancer. Animal studies have shown that inhalation of air particulates induces mutations in the male germline. Expanded simple tandem repeat (ESTR) loci in mice are sensitive markers of mutagenic effects on male germ cells resulting from environmental exposures; however, female germ cells have received little attention. Oocytes may be vulnerable during stages of active cell division (e.g., during fetal development). Accordingly, an increase in germline ESTR mutations in female mice prenatally exposed to radiation has previously been reported. Here we investigate the effects of nanoparticles on the female germline. Since pulmonary exposure to nanosized titanium dioxide (nanoTiO(2)) produces a long-lasting inflammatory response in mice, it was chosen for the present study. FINDINGS: Pregnant C57BL/6 mice were exposed by whole-body inhalation to the nanoTiO(2) UV-Titan L181 (~42.4 mg UV-Titan/m(3)) or filtered clean air on gestation days (GD) 8-18. Female C57BL/6 F1 offspring were raised to maturity and mated with unexposed CBA males. The F2 descendents were collected and ESTR germline mutation rates in this generation were estimated from full pedigrees (mother, father, offspring) of F1 female mice (192 UV-Titan-exposed F2 offspring and 164 F2 controls). ESTR mutation rates of 0.029 (maternal allele) and 0.047 (paternal allele) in UV-Titan-exposed F2 offspring were not statistically different from those of F2 controls: 0.037 (maternal allele) and 0.061 (paternal allele). CONCLUSIONS: We found no evidence for increased ESTR mutation rates in F1 females exposed in utero to UV-Titan nanoparticles from GD8-18 relative to control females.

Pulmonary exposure to carbon black by inhalation or instillation in pregnant mice: effects on liver DNA strand breaks in dams and offspring.

Effects of maternal pulmonary exposure to carbon black (Printex 90) on gestation, lactation and DNA strand breaks were evaluated. Time-mated C57BL/6BomTac mice were exposed by inhalation to 42 mg/m(3) Printex 90 for 1 h/day on gestation days (GD) 8-18, or by four intratracheal instillations on GD 7, 10, 15 and 18, with total doses of 11, 54 and 268 µg/animal. Dams were monitored until weaning and some offspring until adolescence. Inflammation was assessed in maternal bronchoalveolar lavage (BAL) 3-5 days after exposure, and at weaning. Levels of DNA strand breaks were assessed in maternal BAL cells and liver, and in offspring liver. Persistent lung inflammation was observed in exposed mothers. Inhalation exposure induced more DNA strand breaks in the liver of mothers and their offspring, whereas intratracheal instillation did not. Neither inhalation nor instillation affected gestation and lactation. Maternal inhalation exposure to Printex 90-induced liver DNA damage in the mothers and the in utero exposed offspring.

Effects of prenatal exposure to surface-coated nanosized titanium dioxide (UV-Titan). A study in mice.

BACKGROUND: Engineered nanoparticles are smaller than 100 nm and designed to improve or achieve new physico-chemical properties. Consequently, also toxicological properties may change compared to the parent compound. We examined developmental and neurobehavioral effects following maternal exposure to a nanoparticulate UV-filter (UV-titan L181). METHODS: Time-mated mice (C57BL/6BomTac) were exposed by inhalation 1h/day to 42 mg/m(3) aerosolized powder (1.7.10(6) n/cm(3); peak-size: 97 nm) on gestation days 8-18. Endpoints included: maternal lung inflammation; gestational and litter parameters; offspring neurofunction and fertility. Physicochemical particle properties were determined to provide information on specific exposure and deposition. RESULTS: Particles consisted of mainly elongated rutile titanium dioxide (TiO2) with an average crystallite size of 21 nm, modified with Al, Si and Zr, and coated with polyalcohols. In exposed adult mice, 38 mg Ti/kg was detected in the lungs on day 5 and differential cell counts of bronchoalveolar lavage fluid revealed lung inflammation 5 and 26-27 days following exposure termination, relative to control mice. As young adults, prenatally exposed offspring tended to avoid the central zone of the open field and exposed female offspring displayed enhanced prepulse inhibition. Cognitive function was unaffected (Morris water maze test). CONCLUSION: Inhalation exposure to nano-sized UV Titan dusts induced long term lung inflammation in time-mated adult female mice. Gestationally exposed offspring displayed moderate neurobehavioral alterations. The results are discussed in the light of the observed particle size distribution in the exposure atmosphere and the potential pathways by which nanoparticles may impart changes in fetal development.

Lack of acute phase response in the livers of mice exposed to diesel exhaust particles or carbon black by inhalation.

BACKGROUND: Epidemiologic and animal studies have shown that particulate air pollution is associated with increased risk of lung and cardiovascular diseases. Although the exact mechanisms by which particles induce cardiovascular diseases are not known, studies suggest involvement of systemic acute phase responses, including C-reactive protein (CRP) and serum amyloid A (SAA) in humans. In this study we test the hypothesis that diesel exhaust particles (DEP) - or carbon black (CB)-induced lung inflammation initiates an acute phase response in the liver. RESULTS: Mice were exposed to filtered air, 20 mg/m3 DEP or CB by inhalation for 90 minutes/day for four consecutive days; we have previously shown that these mice exhibit pulmonary inflammation (Saber AT, Bornholdt J, Dybdahl M, Sharma AK, Loft S, Vogel U, Wallin H. Tumor necrosis factor is not required for particle-induced genotoxicity and pulmonary inflammation., Arch. Toxicol. 79 (2005) 177-182). As a positive control for the induction of an acute phase response, mice were exposed to 12.5 mg/kg of lipopolysaccharide (LPS) intraperitoneally. Quantitative real time RT-PCR was used to examine the hepatic mRNA expression of acute phase proteins, serum amyloid P (Sap) (the murine homologue of Crp) and Saa1 and Saa3. While significant increases in the hepatic expression of Sap, Saa1 and Saa3 were observed in response to LPS, their levels did not change in response to DEP or CB. In a comprehensive search for markers of an acute phase response, we analyzed liver tissue from these mice using high density DNA microarrays. Globally, 28 genes were found to be significantly differentially expressed in response to DEP or CB. The mRNA expression of three of the genes (serine (or cysteine) proteinase inhibitor, clade A, member 3C, apolipoprotein E and transmembrane emp24 domain containing 3) responded to both exposures. However, these changes were very subtle and were not confirmed by real time RT-PCR. CONCLUSION: Our findings collectively suggest that Sap, Saa1 and Saa3 are not induced in livers of mice exposed to DEP or CB. Despite pulmonary inflammation in these mice, global transcriptional profiling of liver did not reveal any hepatic response following exposure by inhalation.