Potential application of body fluids autofluorescence in the non-invasive diagnosis of endometrial cancer.

BACKGROUND: Endometrial carcinoma (EC) is the most common cancer of the female reproductive tract in developed countries. The prognosis and 5-year survival rates are closely tied to the stage diagnosis. Current routine diagnostic methods of EC are either lacking specificity or are uncomfortable, invasive and painful for the patient. As of now, the gold diagnostic standard is endometrial biopsy. Early and non-invasive diagnosis of EC requires the identification of new biomarkers of disease and a screening test applicable to routine laboratory diagnostics. The application of untargeted metabolomics combined with artificial intelligence and biostatistics tools has the potential to qualitatively and quantitatively represent the metabolome, but its introduction into routine diagnostics is currently unrealistic due to the financial, time and interpretation challenges. Fluorescence spectral analysis of body fluids utilizes autofluorescence of certain metabolites to define the composition of the metabolome under physiological conditions. PURPOSE: This review highlights the potential of fluorescence spectroscopy in the early detection of EC. Data obtained by three-dimensional fluorescence spectroscopy define the quantitative and qualitative composition of the complex fluorescent metabolome and are useful for identifying biochemical metabolic changes associated with endometrial carcinogenesis. Autofluorescence of biological fluids has the prospect of providing new molecular markers of EC. By integrating machine learning and artificial intelligence algorithms in the data analysis of the fluorescent metabolome, this technique has great potential to be implemented in routine laboratory diagnostics.

NK cells not only in endometrium but also in ovulatory cervical mucus in patients with decreased fertility.

OBJECTIVE: An increased number of NK cells is associated with autoimmune disorder and is known to play a role in infertility. The aim of our research was to monitor the density of NK cells CD56+ and CD16+ in ovulatory cervical mucus (OCM) and in endometrium in infertile women as well as in connection with the actual status of antibodies against phospholipids, sperm and HHV-6 antibodies. TYPE OF STUDY: Original aticle. SETTING: Genetika - Plzeň. METHODS: Seventy-two randomly selected women aged 20-39 (mean age: 32.3) years old resulted in fifty-seven patients with repeated unexplained miscarriages, and fifteen fertile healthy women. The hormonal status was studied including ovulation, the humoral autoimmune responses to eight phospholipids, trombophilia, karyotyping, hysteroscopy, and endometrium immunohistology. Patients were without any clinical and laboratory symptoms of vaginitis at the time of OCM sampling and endometrium study. In one patient antiphospholipid syndrome was present, and in one woman diabetes mellitus was identified. Uterine NK cells CD56+ , CD16+ and NK cells in OCM were identified by immunocytochemistry, antiphospholipid antiboides by ELISA. We used indirect MAR-test for study of local spermagglutinating antibodies in OCM. Indirect immunofluorescent method was used for detection of serum and OCM IgM, IgG antibodies against HHV-6 levels at the time of ovulation. RESULTS: We found both high density of NK cells CD56+ and CD16+ in OCM and in endometrium in only two infertile women with repeated abortions. NK cells in OCM were missing in other samples of patients. The prevalence of high density of NK cells CD56+ in the endometrium was seen in twenty three (40%), NK cells CD16+ in eleven (19%), NK cells 56+ and NK cells 16+ together in eight (14%). Levels of serum and OCM IgG against HHV-6 in all examined patients were not elevated, no cervical sperm antibodies were found. CONCLUSION: We compared density of NK cells CD56+ and CD16+ in OCM and secretory endometrium in all infertile patients. Our results show that cell mucosal activity in the cervical area at the time of ovulation in two infertile patients was evident. We excluded the abnormal number of NK cells owing to local and general viral infection (HHV-6). But our question still remains - are cervical NK cells fixed or still migrating from endometrium into OCM? New research is planned.

[Cytological-energetic principle of the amniotic fluid examination on female patients with diagnosis of preterm premature rupture of membranes - up to now experiences and perspectives]. / Cytologicko-energetický princip vysetrování plodové vody po predcasném predtermínovém odtoku - dosavadní zkusenosti a perspektivy.

In our study we have dealt with the amniotic fluid examination on female patients with diagnosis of preterm premature rupture of membranes (PPROM). For one of the most important task in the feto-maternal medicine we consider the discovered method of amniotic fluid examination which is capable of the earliest possible detection in an intraamniotic inflammatory response. Unnoticed could harm or in the most serious cases threat the life of fetus or its mother. This was also the main reason why we chose this topic for our study. We have not been interested only in already known interleu-kin-6 (IL-6), but we have approached the examination comprehensively. We used the cytological-energetic principle supplemented by the examination of aspartate-amino-transferase (AST), in which already are long-term experiences in investigating other extracellular fluids. We have monitored values of C-reactive protein (CRP) and leucocytes in maternal serum, IL-6, AST and energetic score ratio (KEB) in amniotic fluid. Further we have investigated which imunocompetent cells have been dominant in amniotic fluid at different types of inflamantory reaction. According to the results of the examination of the control groups, it was necessary to correct the KEB limit in comparison to KEB limits of other extracellular fluids. Although our study includes untill now only 44 patients - 21 controls and 23 investigated persons, we have been already able to find out some trends of changes in observed parametres. As a very impotant discovery we consider the fact, that KEB, AST and cytological examination of amniotic fluid seem to be the methods for prediction of chorioamnionitis as reliable as the examination of IL-6. These methods are besides less expensive and therefore more suitable for the use in the small hospitals and developing countries. We continue in our study to confirm the previously found trends on the larger numbers of patients.

The diagnosis of norovirus infections in hospitalized children and adolescents with acute gastroenteritis: a study from Pilsen, Czech Republic.

BACKGROUND: Norovirus-associated sporadic gastroenteritis is an important cause of illness in Western Europe. However, at present, little information on the role of norovirus in sporadic gastroenteritis in Central Europe is available. Our study aimed at providing an assessment of their significance in hospitalized children and adolescents with acute gastroenteritis using ELISA test at the time of their introduction. METHODS: A prospective hospital based study of the etiology of acute gastroenteritis was undertaken in a total of 618 patients (mean age 39.8 months, range 0-228), who were hospitalized at the Charles University Hospital in Pilsen. All subjects were monitored in six fragmented periods during the years 2003 and 2004, 2006 and 2007. Clinical and laboratory data were processed, norovirus antigens in stools were detected using the EIA kits IDEIA Norovirus, DakoCytomation. RESULTS: A norovirus infection was confirmed in 62 cases, i.e., 10.0% of all patients with acute gastroenteritis. Vomiting was the most common symptom, recorded in 95.2 % of all the patients with norovirus infection. No severe extragastrointestinal complications were detected. The average interval between initial symptoms and the beginning of hospitalization was considerably shorter in patients with norovirus infection (1.3 days) compared with patients with rotavirus infections (2.4 days). The frequency of Salmonella spp., rotavirus, Campylobacter spp. and enteric adenovirus was 15.4%, 11.2%, 3.9%, 3.6%, respectively. CONCLUSIONS: Our findings confirm the clinical importance of noroviruses as a causative agent of acute gastroenteritis in children and teenagers in the region of a Central European country. Identification of norovirus infection should be included in the routine screenings of sporadic cases of acute gastroenteritis.

Valacyclovir prophylaxis versus preemptive valganciclovir therapy to prevent cytomegalovirus disease after renal transplantation.

Both preemptive therapy and universal prophylaxis are used to prevent cytomegalovirus (CMV) disease after transplantation. Randomized trials comparing both strategies are sparse. Renal transplant recipients at risk for CMV (D+/R-, D+/R+, D-/R+) were randomized to 3-month prophylaxis with valacyclovir (2 g q.i.d., n = 34) or preemptive therapy with valganciclovir (900 mg b.i.d. for a minimum of 14 days, n = 36) for significant CMV DNAemia (>/=2000 copies/mL by quantitative PCR in whole blood) assessed weekly for 16 weeks and at 5, 6, 9 and 12 months. The 12-month incidence of CMV DNAemia was higher in the preemptive group (92% vs. 59%, p < 0.001) while the incidence of CMV disease was not different (6% vs. 9%, p = 0.567). The onset of CMV DNAemia was delayed in the valacyclovir group (37 +/- 22 vs. 187 +/- 110 days, p < 0.001). Significantly higher rate of biopsy-proven acute rejection during 12 months was observed in the preemptive group (36% vs. 15%, p = 0.034). The average CMV-associated costs per patient were $5525 and $2629 in preemptive therapy and valacyclovir, respectively (p < 0.001). However, assuming the cost of $60 per PCR test, there was no difference in overall costs. In conclusion, preemptive valganciclovir therapy and valacyclovir prophylaxis are equally effective in the prevention of CMV disease after renal transplantation.

[Diagnosis of rotavirus infections--comparison of various methods]. / Diagnostika rotavirových infekcí--porovnání ruzných metod.

In 179 samples of faeces from patients hospitalized with diarrhoeal diseases rotavirus infections were examined concurrently by three different sets--the latex agglutination test Diarlex Rota-Adeno (Orion Diagnostica) and two immunoenzyme tests (EIA)--EIA Rotavirus (Test-Line, CR) and IDEIA Rotavirus (Dako Ltd.) By the latex agglutination reaction the rotavirus infection was proved 31 times (17.3%), by the Test-Line test 87 times (48.6%), by the Dako test 96 times (53.6%). During the interval of 0-3 days after the onset of the disease the positivity assessed by different tests was 22.8%, 48.6% and 54.3%, on the 4th-7th day 12.8%, 51.1% and 57.4%, on the 8th and subsequent days 3.7%, 44.4% and 44.4%. Comparison of sets Orion Diagnostica and Dako revealed the sensitivity of the latex agglutination test only as 30.2% and specificity 97.6%. After the 9th day from the onset of the disease the infection was no longer confirmed by the latex agglutination test, while both EIA tests proved the rotavirus antigen up to the 16th day from the onset of the disease.

[Monitoring cytomegalovirus infection using the 2-step nested polymerase chain reaction in patients with allogenic bone marrow transplantation]. / Monitorace aktivity cytomegalovirové infekce u nemocných po alogenní transplantaci kostní drene metodou dvoustupnové "nested" polymerázové retezové reakce.

Cytomegalovirus infections and their sequelae are the most serious complications in patients after allogenic bone marrow transplantations. Therefore in recent years quite rightly attention is paid to new diagnostic methods which make earlier and more sensitive detection of incipient CMV infection possible. The method of the two-stage polymerase chain reaction (PCR) was used for early detection of CMV infection and its possible further monitoring in 38 recipients of allogenic bone marrow. A positive result was recorded in 15 (39%) patients whereby in 10 (13%) repeated positivity was involved. In three in this group of patients (8%) CMV disease developed, always preceded by PCR positivity. Sensitivity, specificity, positive and negative predictive CMV PCR values were 100, 66, 20 and 100%. Concomitant comparison with serological examination (antibody titre class IgM/seroconversion) revealed the small contribution of serology (corresponding values are 67, 54, 11 and 95%) in patients after bone marrow transplantations. The presented results provide evidence of the contribution of PCR in the diagnosis of CMV in those patients where due to their high sensitivity and excellent predictive value it is possible to avoid the toxicity of unnecessary antiviral treatment.