Prenatal ultrasonic detection of bilateral hydrocele.
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Adult↗
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The acquisition of multidimensional NMR spectra within a single scan.
A scheme enabling the complete sampling of multidimensional NMR domains within a single continuous acquisition is introduced and exemplified. Provided that an analyte's signal is sufficiently strong, the acquisition time of multidimensional NMR experiments can thus be shortened by orders of magnitude. This could enable the characterization of transient events such as proteins folding, 2D NMR experiments on samples being chromatographed, bring the duration of higher dimensional experiments (e.g., 4D NMR) into the lifetime of most proteins under physiological conditions, and facilitate the incorporation of spectroscopic 2D sequences into in vivo imaging investigations. The protocol is compatible with existing multidimensional pulse sequences and can be implemented by using conventional hardware; its performance is exemplified here with a variety of homonuclear 2D NMR acquisitions.
Animals↗
CYP1A1 and GSTM1 genetic polymorphisms in lung cancer populations exposed to arsenic in drinking water.
Region II of Chile is the most important copper mining area in the world and it shows the highest lung cancer mortality rate in the country (35/100,000). The population in Antofagasta, the main city of Region II, was exposed from 1958 to 1970 to 860 microg m(-3) arsenic (As) in drinking water and has currently been declining to 40 microg m(-3). Glutathione serves as a reducing agent and glutathione S-transferase (GST) may have an important role in As methylation capacity and body retention. In the current study, the null genotype of GSTM1 and the MspI polymorphism of CYP450 1A1 were investigated in lung cancer patients and in healthy volunteers of Region II. In males, the 2A genotype of MspI represented a highly significant estimated relative lung cancer risk (OR=2.60). Relative lung cancer risk for the combined 2A/null GSTM1 genotypes was 2.51, which increased with the smoking habit (OR=2.98). In Region II, the cancer mortality rate for As-associated cancers at least partly might be related to differences in As biotransformation. Genetic biomarkers such as 2A and GSTM1 polymorphisms in addition to DR70 as screening biomarkers might provide relevant information to identify individuals with a high risk for lung cancer as prevention and protection actions to protect public health.
Arsenic↗
Hepatic enzyme induction and mutagenicity of airborne particulate matter from Santiago, Chile in the nourished and malnourished rat.
1. Respirable, airborne particles in the ambient air in downtown Santiago, Chile, have been characterized for the seasonal variation in total polycyclic aromatic hydrocarbon content, 13 of which have been identified including the mutagens (benzo(a)pyrene, dibenzo(a,h)anthracene, benzo(a)anthracene, benzo(b)fluoranthene and indeno(1,2,3, c,d)pyrene amongst others. 2. Organic extracts derived from these particles were administered to both the nourished and malnourished rat and resulted in modulation of the hepatic mixed function oxidase system including induction of NADPH-cytochrome P450 reductase, cytochrome P4501A as determined by Western blot analysis and the associated ethoxyresorufin O-deethylase and aryl hydrocarbon hydroxylase activities. 3. The cytochrome P4504A1-dependent 12-hydroxylation of lauric acid was induced in the malnourished state, but this activity was significantly inhibited by treatment of the animals with particle extracts in both nutritional states. 4. The particle extracts contained both direct and indirect-acting mutagens in the Ames test, and depending on the relative complement of both, resulted in either increased or decreased mutagenicity in the presence of S9 activation systems derived from both nourished and malnourished animals. 5. These results are discussed in the context of the interindividual risk assessment of airborne, particulate matter to man.
Air Pollutants↗
Polycyclic aromatic hydrocarbon levels and mutagenicity of inhalable particulate matter in Santiago, Chile.
The air in Santiago, Chile, is among the most highly polluted in the world. Due to the high levels of pollutants and the high incidence of respiratory diseases, especially in the most susceptible groups, Santiago has been declared a saturated zone for PM(10), O(3), and CO. The aim of this work was to investigate polycyclic aromatic hydrocarbon levels and mutagenic activity of Santiago s fine and coarse fractions of inhalable particles. The levels of 16 polycyclic aromatic hydrocarbons (PAHs) were determined by high-performance liquid chromatography in organic extracts from respirable particles (OERP). Respirable particulate matter (fine and coarse) contains high levels of PAHs including six mutagenic ones classified by the IARC as carcinogenic, which represented at least 45% of the total PAH concentration. A seasonal effect was observed, with higher values in months with lower temperatures. Although a significant decline of PAH levels in OERP was observed in the last years, the levels of carcinogenic PAHs are still higher than those reported in cities of the United States, Australia, and Europe. OERP were highly mutagenic and contained direct and indirect mutagens, which produced both frameshift and base substitution mutations in Salmonella typhimurium. In addition, organic extracts from total suspended particles were also highly mutagenic at the tk locus in h1A1v2 human lymphoblasts in culture. In spite of the important decrease in PAHs in the period 1991-1996, direct mutagenic response has not changed significantly, suggesting that the levels of direct mutagenic pollutants (e.g., nitroarenes) have not decreased considerably during the last years. These results suggest a risk for Santiago s inhabitants since pollutants adsorbed in inhalable particles are highly mutagenic and can damage DNA.
Administration, Inhalation↗
Trends of polycyclic aromatic hydrocarbon levels and mutagenicity in Santiago's inhalable airborne particles in the period 1992-1996.
Trends of polycyclic aromatic hydrocarbons (PAHs) for 1992-1996 (cold season) and their mutagenic activity were investigated in organic extracts from the Santiago, Chile, inhalable particles (PM(10)). The highest PAH concentrations were observed in 1992 and declined dramatically in the following years. During this period, total PAHs decreased 85%, carcinogenic PAHs 82%, and benzo[a]pyrene, the most potent carcinogen, 85%. In spite of this significant decrease, PAH levels in respirable particles were higher than those reported in recent studies in Australia, Europe, and the United States. PAH profiles were analyzed by principal component (PC) analysis and Pearson correlation analysis. PC1 represents 71% of the variance, suggesting that most PAHs might originate predominantly from one main generic source. Higher correlations were obtained for the major carcinogenic PAHs. Most of the samples assayed were highly mutagenic to Salmonella typhimurium both in the presence and in the absence of metabolic activation system (S9), especially in the coarse fraction, but direct mutagenicity did not decline significantly. Incubation of calf thymus DNA with organic extracts from particulate matter and xanthine oxidase allowed the detection of five nitro-PAH-DNA adducts. Thus, nitroarenes might play an important role in the mutagenic activity of inhalable particles in Santiago, representing a high risk for human health.
Administration, Inhalation↗
Quantification of viral inactivation by photochemical treatment with amotosalen and UV A light, using a novel polymerase chain reaction inhibition method with preamplification.
BACKGROUND: In evaluating a photochemical treatment process for inactivating parvovirus B19, there lacked simple culture methods to measure infectivity. The recently developed enzyme-linked immunospot (ELISpot) infectivity assay uses late-stage erythropoietic progenitor cells and is labor intensive and time consuming. We evaluated a novel, efficient polymerase chain reaction (PCR) inhibition assay and examined correlations with reductions in infectivity. METHODS: Contaminated plasma was treated with 150 micromol/L amotosalen and 3 J/cm(2) ultraviolet A light and then tested for DNA modification using conventional PCR inhibition and a novel preamplification approach. The novel assay subjected the samples to preamplification cycles using long-template PCR, followed by quantitative PCR (QPCR) inhibition detection. Both approaches were tested for correlations with reductions in viral infectivity by comparing ELISpot assay results of identical samples. RESULTS: The B19 preamplification inhibition assay showed detection ranges of 2-2.5 log and demonstrated quantitative correlation with up to a 5.8-log reduction in viral infectivity in ELISpot results. Conventional PCR detected a >5 log reduction in amplification, correlated with a 4.4-log reduction in viral infectivity. A range of 4-log inhibition of hepatitis B virus DNA amplification was also achieved. CONCLUSIONS: The results demonstrated that a novel preamplification QPCR assay is a useful tool for predicting reductions in infectivity after photochemical treatment. This assay was extended to show utility in circumstances where practical in vitro assays are unavailable for the determination of the efficacy of pathogen inactivation.
DNA, Viral↗
Identification of novel inhibitors of Mycobacterium smegmatis growth through genome-wide overexpression of Cluster P3 mycobacteriophage Xavia genes.
Bacteriophages encode numerous genes with no known function, many of which can affect essential cellular processes when expressed in the bacterial host. For mycobacteriophages, genome-wide overexpression in Mycobacterium smegmatis can be used to identify proteins that impair growth. To evaluate the cytotoxic potential of the Cluster P3 phage Xavia, we constructed a plasmid library containing 71 predicted Xavia genes under the anhydrotetracycline inducible promoter pTet and screened this library in a plate-based cytotoxicity assay to measure impacts on M. smegmatis growth. Two genes prevented transformants recovery, consistent with toxicity under basal promoter leakiness, and inducible expression of 18 additional genes impaired growth. These inhibitory proteins include structural components; factors involved in DNA metabolism, lysogeny, and lysis; and several proteins with no known function. These results extend functional screening into a lineage of actinobacteriophages that has not previously been characterized, and identify new proteins that warrant further mechanistic analysis.
Mycobacterium smegmatis↗
Intraperitoneal levonorgestrel-releasing intrauterine device following uterine perforation: the role of progestins in adhesion formation.
BACKGROUND: Intrauterine contraception is a widely used, highly effective means of birth control. Uterine perforation is a serious, albeit rare, complication of intrauterine device (IUD) use. Although uterine perforation by levonorgestrel-releasing (20 micro g/day) intrauterine system (LNG-IUS) has already been reported, the peritoneal adhesion potential of this IUD is unknown. METHODS: The medical files of all patients diagnosed with an intra-peritoneal IUD between the years 1990-2002 at Hadassah Medical Center were reviewed. Histopathological study of peritoneal adhesion tissue adjacent to levonorgestrel medicated IUD was conducted in one case. RESULTS: Eight cases of dislocated IUDs were found. Four cases used LNG-IUS and four other cases used copper-IUD. Laparoscopy for IUD removal disclosed mild local peritoneal adhesions between omentum and pelvic organs in all cases. No difference was noted in the appearance of the peritoneum in the presence of either a copper-IUD or LNG-IUS. Histological examination of peritoneal tissue encasing the levonorgestrel-intrauterine system revealed loose connective tissue with aggregates of submesothelial cells with a pseudo-decidual change. Immunohistochemical staining for progesterone receptor was negative. CONCLUSIONS: The peritoneal adhesions potential of LNG-IUS is low, similar to that of the copper-bearing IUD.
Adult↗
Management of a perforated levonorgestrel-medicated intrauterine device--a pharmacokinetic study: case report.
Intrauterine contraception is a widely used, highly effective method of birth control. Uterine perforation is a serious albeit rare complication with the use of an intrauterine device (IUD). Although uterine perforation by the levonorgestrel-releasing intrauterine system (LNG-IUS) has already been described, no plasma LNG concentrations in this setting were reported. Neither has the management of LNG-IUS been commented on to date. Two months after insertion of an LNG-IUS into a 33-year-old woman, it was noted to be in the peritoneal cavity. Laparoscopy for IUD removal was conducted 5 months after insertion. LNG and sex hormone-binding globulin plasma concentrations were measured prior to and following the laparoscopic removal of the IUD. Intra-peritoneal dislocated LNG-IUS resulted in plasma LNG levels 10 times higher (4.7 nmol/l) than the plasma level of LNG observed with LNG-IUS placed in utero. This high plasma LNG level suppresses ovulation. Therefore a misplaced LNG-IUS should be removed when pregnancy is desired.
Adult↗
A survey of splice variants of the human hypoxanthine phosphoribosyl transferase and DNA polymerase beta genes: products of alternative or aberrant splicing?
Errors during the pre-mRNA splicing of metazoan genes can degrade the transmission of genetic information, and have been associated with a variety of human diseases. In order to characterize the mutagenic and pathogenic potential of mis-splicing, we have surveyed and quantified the aberrant splice variants in the human hypoxanthine phosphoribosyl transferase (HPRT) and DNA polymerase beta (POLB) in the presence and the absence of the Nonsense Mediated Decay (NMD) pathway, which removes transcripts with premature termination codons. POLB exhibits a high frequency of splice variants (40-60%), whereas the frequency of HPRT splice variants is considerably lower (approximately 1%). Treatment of cells with emetine to inactivate NMD alters both the spectrum and frequency of splice variants of POLB and HPRT. It is not certain at this point, whether POLB and HPRT splice variants are the result of regulated alternative splicing processes or the result of aberrant splicing, but it appears likely that at least some of the variants are the result of splicing errors. Several mechanisms that may contribute to aberrant splicing are discussed.
Alternative Splicing↗
Human herpesvirus 6 infection mimicking juvenile myelomonocytic leukemia in an infant.
In vitro cell culture studies of bone marrow and peripheral blood progenitor cells from patients with juvenile myclomonocytic leukemia (JMML) consistently show spontaneous proliferation and selective hypersensitivity to granulocyte-macrophage colony-stimulating factor (GM-CSF). This GM-CSF hypersensitivity dose-response assay has become a component of the international diagnostic criteria for JMML. The authors report a 2-week-old boy with perinatal human herpesvirus 6 (HHV-6) infection in whom in vitro bone marrow culture studies suggested the diagnosis of JMML by showing increased spontaneous proliferation, inhibition of this growth by anti-GM-CSF antibodies, and hypersensitivity to GM-CSF. Polymerase chain reaction viral studies from whole blood DNA and the shell vial viral culture assay were both positive for HHV-6. The patient's condition improved with expectant treatment, with an eventual return to normal blood counts and resolution of hepatosplenomegaly. This case of perinatal HHV-6 infection shows that viruses can initially mimic the in vitro culture results found in patients with JMML. It also illustrates that patients suspected of having JMML should be observed if there are no signs of progressive disease and concurrent features suggestive of viral infection.
Acute Kidney Injury↗
Reliability and agreement of urodynamics interpretations in a female pelvic medicine center.
OBJECTIVE: To estimate the reliability and interobserver consistency of urodynamic interpretations of female bladder and urethral function. METHODS: Three urogynecologists and three female urologists at a tertiary care medical center reviewed masked, abstracted clinical and urodynamic information from 100 charts, selected for adequate completeness from a consecutive series of 135 women referred for urodynamic testing. For each of the 100 cases, the reviewers assigned International Continence Society filling and voiding phase diagnoses, and overall clinical diagnoses. Raw agreement proportions and weighted kappa chance-corrected agreement statistics (kappa) were used jointly to describe both reliability and interobserver agreement. Reliability was estimated from duplicate reviews, masked and separated by at least 4 months, of each case by each physician. Interobserver agreement was estimated from comparisons of all pairs of responses from different physicians. RESULTS: For clinical diagnosis of stress incontinence (present, absent, indeterminate), the within- and across-physician weighted kappa's were, respectively, 0.78 and 0.68. Corresponding results were 0.40 and 0.13 for detrusor overactivity without incontinence, 0.58 and 0.38 for detrusor overactivity with incontinence, and 0.51 and 0.26 for voiding dysfunction. Standard errors of each kappa were between 0.023 and 0.043. CONCLUSION: In our group, lower urinary tract diagnoses of stress urinary incontinence from both clinical and urodynamic data demonstrated substantial reliability and interobserver agreement. However, by conventional interpretation of kappa-statistics, reliability of diagnoses of detrusor overactivity or voiding dysfunction was only moderate, and interobserver agreement on these diagnoses was no better than fair. Urodynamic interpretations may not be satisfactorily reproducible for these diagnoses.
Diagnostic Techniques, Urological↗
Occupational and environmental levels of mutagenic PAHs and respirable particulate matter associated with diesel exhaust in Santiago, Chile.
This work studied the mutagenic potential and polycyclic aromatic hydrocarbon (PAH) levels onto PM10 collected in diesel revision plants, in an urban area as well as in a rural area in Santiago, Chile. The PM10 average levels in diesel emission plants during working hours (there is no occupational PM10 Chilean standard) were significantly higher than the atmospheric Chilean PM10 standard and highly mutagenic and with high PAHs levels. Additionally, we evaluated the contribution of CYP1A1 and GSTM1 polymorphisms on 1-OH-pyrene urinary levels. The diesel-exposed workers carrying the CYP1A1*2A allele showed significantly higher 1-OH-P levels than the subjects from the rural area with the same genotype. The higher levels of 1-OH-P were found in individuals carrying the combined CYP1A1*2A and GSTM1 null genotype. This kind of information might be relevant to establish prevention, protection, and mitigation actions to protect public health.
Adult↗
Efficacy of a vaginal sling procedure in a rat model of stress urinary incontinence.
PURPOSE: We validated the efficacy of the vaginal sling procedure for the restoration of leak point pressure (LPP) in the rat model of stress urinary incontinence (SUI). MATERIALS AND METHODS: SUI was created in 10 female Sprague-Dawley rats by bilateral pudendal nerve transection (PNT) under urethane anesthesia. Vaginal dissection was performed, followed by placement of a 2 x 0.3 cm strip of Prolene (Ethicon, Somerville, New Jersey) mesh at the mid urethral level. LPP was measured before and after PNT through a suprapubic tube using a Credé maneuver. It was also measured after vaginal dissection (sham sling) and after true sling placement. All procedures and measurements were done at the same experimental setting. In each animal LPP was measured 4 or 5 times and the mean was taken. Pairwise differences in LPP between the true and sham slings were calculated using the Wilcoxon signed rank test with p <0.05 considered significantly different. RESULTS: Mean LPP at baseline in all rats was 65.1 +/- 6.0 cm water. LPP decreased to 42.7 +/- 3.7 cm water after PNT. The sham sling further decreased LPP to 26.5 +/- 2.2 cm water, whereas the true sling restored LPP to 56 +/- 4.8 cm water. LPP after true sling placement was not different from baseline (p = 0.16), whereas LPP after sham sling placement was significantly different from baseline (p = 0.002). CONCLUSIONS: This rat vaginal sling model represents a valid surgical method for LPP restoration in the rat model of SUI. This animal model could be used for future studies related to SUI and its treatment relevant to sling surgery.
Animals↗
Long-term efficacy of a vaginal sling procedure in a rat model of stress urinary incontinence.
PURPOSE: We examined the long-term efficacy of a newly created vaginal sling procedure for the restoration of leak point pressure (LPP) in a rat model of stress urinary incontinence (SUI). MATERIAL AND METHODS: A total of 20 female Sprague-Dawley rats were randomly assigned to 1 of 4 groups, namely normal control, SUI plus vaginal sling, SUI plus sham sling and SUI only. SUI was created in the latter 3 groups by bilateral pudendal nerve transection (PNT). In the sling procedure a 2 x 0.3 cm strip of polypropylene mesh was placed at the mid urethral level. Animals in the SUI plus sham sling group underwent vaginal dissection only. After 5 weeks LPP was measured 4 or 5 times in each rat and the mean was determined. The Kruskal-Wallis and Wilcoxon rank sum tests were used to evaluate whether levels of measurements differed across and between groups. RESULTS: Mean LPP +/- SD in control rats was 48.8 +/- 10.2 cm H2O. PNT decreased LPP to 23.5 +/- 7.4 cm H2O. Sling placement improved LPP at 5 weeks to 35.5 +/- 2.3 cm H2O, whereas LPP in the sham sling group was 29.1 +/- 4.9 cm H2O. LPP recovery in the sling group was significantly above levels for PNT (p = 0.037). LPP in the sling group did not differ statistically from that in the control group (p = 0.11). CONCLUSIONS: The newly created vaginal sling model restores the LPP in the rat model of SUI in the long term (5 weeks). This model could be used to address research questions related to the sling procedure.
Animals↗
Role of sling integrity in the restoration of leak point pressure in the rat vaginal sling model.
PURPOSE: We tested the hypothesis that cutting the sling at its suburethral section does not cancel its anti-incontinence effect. We also examined the long-term effects of the sling on bladder function in a recently validated rat model of vaginal sling. MATERIALS AND METHODS: Stress urinary incontinence was created in 60 female Sprague-Dawley rats by the previously established method of bilateral pudendal nerve transection. Under anesthesia 20 animals received a vaginal sling, 20 received a vaginal sling in which the suburethral portion of the sling was cut immediately after placement and 20 received a sham vaginal sling. Six weeks after the procedures leak point pressure was determined and a cystometrogram was done using anesthesia in each animal via a previously implanted suprapubic catheter. Kruskal-Wallis and pairwise separate rank multiple comparison tests were performed with a significance level of 0.05. RESULTS: The cut and intact slings increased leak point pressure similarly and these values were significantly higher than that of the sham sling (24.9 and 27.9 cm H2O, respectively, vs 20.7, p <0.0001). Peak micturition pressure was not significantly different among the 3 groups, indicating absent bladder outlet obstruction in the sling groups. Bladder compliance was significantly decreased 6 weeks after placement of a cut or intact sling compared with the sham sling (p = 0.007 and 0.05, respectively). CONCLUSIONS: An intact suburethral portion is not a requirement for sling effectiveness in the rat model of stress urinary incontinence. However, the sling procedure decreases bladder compliance. This may explain the observed voiding dysfunction associated with sling procedures.
Animals↗
Density differences in the spermatozoa of normozoospermic, asthenozoospermic and oligo-asthenozoospermic males.
The spermatozoa densities of the ejaculates obtained daily for five days from infertile males were compared to the spermatozoa densities of the ejaculates obtained from normozoospermic donors. The density distribution of the spermatozoa was established on Percoll columns of discontinuous densities. The density dispersion patterns of five consecutive days from each donor within a donor group were used to establish the main, daily density dispersion patterns. The five, daily, mean dispersion patterns were used to create 3-dimensional graphs of density dispersion variations. The 3-dimensional graphs from the infertile male groups show negligible daily variations as compared to that of the healthy males. The absence of wide daily variations is suggested to be considered as a sign of faulty epididymal functioning.
Centrifugation, Density Gradient↗