Ultrafast laser diode thermal desorption method for analysis of representative pharmaceuticals in soil leachate samples.

We developed and evaluated a novel analytical method combining ambient ionization technique - laser diode thermal desorption with chemical ionization (LDTD-APCI) and tandem mass spectrometry detection. The LDTD/APCI-MS/MS method was developed for determination of representative pharmaceuticals from different classes (carbamazepine, sulfamethoxazole, irbesartan, fexofenadine) in leachate samples from soil sorption experimentation. We then optimized laser pattern, laser energy and spiked sample volume, which are crucial parameters for this LDTD/APCI-MS/MS method. We further identified utility of a chelating agent (Na2-EDTA) to obtain the highest achievable and reproducible signal of target analytes. Achieved method performance parameters (LODs, LOQs, trueness and precision) were comparable with those obtained from LC-MS/MS. However, application of this novel LDTD/APCI-MS/MS method reduced analysis time by two orders of magnitude (to 12 s), compared to more conventional LC-MS/MS approaches, without use of organic solvents. We expect this novel method will reduce costs and increase throughput for future analyses of pharmaceuticals in the environment while advancing a timely principle of green chemistry.

Predictors of local failure in early laryngeal cancer.

The aim of the study was to assess the impact of factors that could predict the probability of local failure in early laryngeal squamous cell carcinoma treated with curative radiotherapy. Sixty seven patients (12 women and 55 men) with laryngeal cancer stage I (47 patients) and stage II (20 patients) were treated from 1998 to 2003 with curative radiotherapy and retrospectively evaluated. Median follow-up was 36 months (3-80). Local relapse occurred in 10 patients (15%), regional lymphnodes relapse affected 2 patients. The median time between start of radiotherapy to recurrence was 13 months (3-48). Death due to cancer occurred in 4 patients (2 died from locoregional progression of the recurrence and 2 from distant metastases), whereas 7 patients died from non-cancer related causes. The 2-year overall survival rate was 90% and 5-year OS was 79%. The 2-year local control rate was 82% and 5-year local control was 79%. In the univariate analysis there was a statistically significant decrease in local control influenced by grading (p < 0.0001). High risk group of relapse encompassed patients with at least two negative factors: supraglottic tumor, women, radiotherapy prolongation by 3 or more days and high grade tumor and has 3 times worse local control than low risk group (p=0.0125). The highest risk of local recurrence was in the first three years after radiotherapy than later (p=0.0057). On multivariate analysis unfavourable prognostic factors for local control were gender (p=0.022), presence of 2 or more negative risk factors (p=0.018) and lengths of follow up (p=0.005). Radiation dose, stage, age, hemoglobin level and anterior commissure involvement were not significant factors for local control. Overall survival was affected both in the univariate and multivariate analysis by presence of local relapse (p < 0.005) and follow up duration (p < 0.02). Anemia had borderline significance for overall survival in univariate analysis (p=0.064), but in the multivariate analysis was significant unfavourable factor (p=0.008). Other studied factors (radiation dose, anterior commissure involvement and age) were not reaching level of statistical significant value for overall survival. Close follow up strategy is recommended for high risk group of patients with two or more risk factors especially in the first three years after radiation therapy.

[Significance of determination of intra-acrosomal proteins and sperm antibodies in human reproduction]. / Význam urcování intra-akrosomálních proteinu a protilátek proti spermiím v lidské reprodukci.

OBJECTIVE: Comparison of the positive intra-acrosomal proteins and spermagglutinating antibodies in human semen samples from various groups of patients. DESIGN: Prospective study. SETTING: Department of Gynecology and Obstetrics and Faculty Hospital, Charles University, Pilsen, Institute of Molecular Genetics, Czech Academy of Science, Prague. METHODS: Monoclonal antibodies Hs-8 and Hs-14 (prepared in the Institute of Molecular Genetics, Prague) were used for detection of intra-acrosomal sperm proteins. Microscopic immunofluorescent methods detected the incidence, the character and the percentage of the spermatozoa specified by above-mentioned monoclonal antibodies. Direct mixed anti-immunoglobulin reactions test (MAR-test) for IgG, IgM, IgA, IgE was used for detection of spermagglutinating antibody. We examined 315 infertile patients from Special Consultation for Immunology of Reproduction and from the IVF programme, and sperm healthy donors (January 2002-March 2003). RESULTS: Native donor's sperm cells had excellent positive intra-acrosomal proteins stained with monoclonal antibodies Hs8 and Hs14 and after thawing as well as. No spermagglutinating antibodies were found. In the group with normal sperm count and light microscopic morphology we found the presence of seminal spermagglutinating antibodies in 11% (IgG), in 14.5% (IgA), in 3.6% (IgM), in 5.2% (IgE). Significant positivity of intra-acrosomal protein stained with Hs8 monoclonal antibody was reached in 68.4%, and with Hs14 monoclonal antibody in even 81.3% of men. On the other hand, in oligoasthenospermatic patients we found significant increasing of spermagglutinating antibodies (for IgG 40.5%, for IgA 28.6%, for IgM 9.5%, for IgE 11.9%). Dominant good staining of intra-acrosomal proteins were seen only in 15.5% of men (for Hs8) and in 20.2% (for Hs14). CONCLUSION: The quantitative detection of intra-acrosomal sperm proteins and spermagglutinating antibodies are used as important properties of human semen and serve for evaluation of acrosomal state, and male fertility together.

[Antizonal antibodies in ovulatory cervical mucus and in serum of patients with fertility disorders]. / Antizonální protilátky v ovulacním hlenu a v séru pacientek s poruchami reprodukce.

OBJECTIVE: Detection of antizonal antibodies (aZPs) in cervical ovulatory mucus and in serum in patients with fertility failure. DESIGN: Retrospective study. SETTING: Department of Gynaecology and Obstetrics, Medical Faculty of Charles University, and Faculty Hospital, Plzen. METHODS: Detection of aZPs (passive hemagglutination test) in cervical ovulatory mucus (OM) and in serum from 105 patients with reproductive failure and in 25 fertile healthy women (control group). RESULTS: Positive levels of aZP in OM were found in 79 (75.2%) patients, in 56 of them aZP were observed in OM and in serum together, only ovulatory aZPs in 23 (21.9%), and only serum aZPs in 18 (17.1%) were found. aZPs predominate in 47 patients (of 48 women) with more than two in vitro fertilization processes, in thirty of them aZPs in serum and in OM together were observed. On the other hand, repeated spontaneous miscarriage and obesity have no connection with detection of aZPs in our patients. CONCLUSION. The increased levels of serum and OM aZPs are very often seen in patients after repeated unsuccessful IVF. This very important result in reproductive immunology means to go on the micromanipulation technics combined with immunosuppression in the next planned IVF procedure.

Gestational progesterone suppresses embryotoxic action of the complement system to chick embryo.

OBJECTIVE: In this paper the relation between progesterone levels and embryotoxic effect of serum complement was studied. DESIGN: The aim of this study was to validate hypothesis that progesterone is strong inhibitor of complement embryotoxic action. SETTING: We used chick embryo like an experimental model for evidence of our hypothesis. We treated chick embryos by sera acquired from healthy pregnant woman with physiologically elevated levels of progesterone and normal complement activity. We investigated embryotoxicity of these sera. RESULT: We noticed a significant decrease of sera embryotoxicity inversely related to serum levels of progesterone. The main finding: The embryotoxicity of sera is reversally dependent on progesterone level. CONCLUSIONS: These findings bring a new knowledge to the role of progesterone and complement system mainly in initial stages of pregnancy and in some cases of spontaneous abortions.