[Basic principles of diagnosis and treatment of secondary peritonitis - recommendations of experts with the support of SIS]. / Základní principy diagnostiky a lécby sekundární peritonitidy - doporucení odborníku s podporou SIS.

Secondary peritonitis is the most common cause of inflammatory acute abdomen treated at general surgery departments. Only early and correct diagnosis may improve the prognosis of these patients. The authors compiled an interdisciplinary review of the basic principles of diagnosis and treatment of secondary peritonitis, which reflects current findings supported by evidence-based medicine. The work is based on published international literature but also shares opinions and experiences of the selected specialists. The presented work in its extent is not meant to substitute an in-depth study of the issue, but to allow a basic and quick review of the topic.

[High sensitivity cardiac troponin assays 2009: clinical potential, current practice and benefits, the future]. / High-sensitivity metody stanovení srdecních troponinu 2009: klinický potenciál, soucasná praxe a prínos, perspektiva.

At present, determination of cardiac troponins (cTn) is the biomarker method of choice for diagnostics and risk stratification in patients with a myocardial injury. Past clinical practice had provided sound evidence that low cTn concentrations, measured with unacceptable imprecision by the currently used methods, hold important clinical, diagnostic and stratification potential. The new generation cTn assays, so called high-sensitivity assays, enable determination of very low cTn concentrations with satisfactory analytical precision and open the way to early identification of small but often prognostically important myocardial damage. Introduction of high-sensitivity cTn assays in practice is, however, associated with some difficulties: their superior diagnostic sensitivity to identify small injuries to myocardium is often linked to lower specificity, higher incidence of elevated cTn concentrations is frequently associated with less obvious clinical symptomatology (overdiagnosis), resulting in greater demand for further patient assessment (overcrowding), repeated analyses and trend monitoring of cTn fluctuation. These initial difficulties cannot lessen the by now indisputable, established benefit of high-sensitivity cTn assays that we briefly describe in the present paper.

Distribution of KIR genes in the Czech population.

Killer cells immunoglobulin-like receptors (KIRs) are a family of inhibitory and activating receptors expressed mainly by natural killer (NK) cells and few subsets of T lymphocytes. KIRs regulate NK cells' activity through interactions with specific HLA class I molecules and other yet unknown ligands presented on target cells. At present, 17 KIR genes and pseudogenes have been identified. As the number of KIR genes in different haplotypes varies, a wide range of genotypes in different ethnic populations may be observed. In our study, 125 healthy non-related Czech individuals were KIR typed both by sequence-specific primers and by sequence-specific oligonucleotide KIR genotyping methods. Thirty-eight different genotypes were observed in the Czech population and all 16 KIR genes known to date were found. Framework genes KIR 3DL3, KIR 2DL4, KIR 3DL2 and the pseudogene KIR 3DP1 were present in all individuals. The most frequent non-framework KIR genes detected in the Czech population were: KIR 2DL1 (95%), KIR 3DL1 (94%), KIR 2DS4 (92%) and the pseudogene 2DP1 (94%). Human leucocyte antigen (HLA)-C typing demonstrated prevalence of the C1/C2 heterozygosity (43%) and C1 homozygosity (41%) over the C2 heterozygosity. One hundred and twenty individuals from our panel carried at least one inhibitory KIR for the corresponding HLA-C group found in the genotype. Gene frequencies and found genotypes demonstrated similarity of the Czech population's KIR repertoire with the KIR repertoires of other Caucasian populations studied before.

A new surface laboratory for measurements of low-level radioactivity.

A laboratory for measuring low-level radioactivity has been built at the AlbaNova University Center in Stockholm. Low-level concrete in combination with a 5 cm iron lining provide an average photon fluency rate from terrestrial sources at least a factor of 40 below that in the common laboratories of the Center. The radon activity inside the laboratory has been measured to (3+/-2) Bq m(-3). Material considerations are discussed. Construction and performance of the laboratory, as well as the present instrumentation, is described.

Construction and biological properties of yellow fever 17D/dengue type 1 recombinant virus.

Dengue virus, a mosquito-borne flavivirus, is one of the most formidable public health threats in tropical and subtropical regions. As yet, there is no licensed vaccine to protect against the disease. A chimeric yellow fever (YF) 17D/dengue (DEN) type 1 virus was constructed by replacing the pre-membrane and envelope genes of YF 17D virus with those from DEN 1 VeMir95 virus, a Venezuelan isolate. The chimeric YF 17D/DEN 1 VeMir95 virus was regenerated from full-length infectious clones stably propagated in Escherichia coli by transfection of Vero cells with in vitro transcribed RNA. The chimeric virus proliferated efficiently in Vero cells ( approximately 6.6 log(10) plaque-forming units/ml). The chimeric virus was not neurovirulent to 3-week-old Swiss Webster mice inoculated by the intracerebral route, in contrast to the YF 17DD vaccine strain that was lethal for 90% of the mice. The YF 17D/DEN 1 virus at Passage 6 was more attenuated for rhesus monkeys than the YF 17DD commercial vaccine after intracerebral inoculation according to the standard neurovirulence test. This virus is a potential candidate to be included in a tetravalent DEN vaccine formulation. The availability of the cloned cDNA allows further structure/function studies on the viral envelope.

[Cardiovascular risk and estrogens]. / Kardiovaskulární riziko a estrogeny.

OBJECTIVE: To describe relations among postmenopause, hormonal therapy, lipid metabolism and risk of cardiovascular diseases. DESIGN: Search and analysis of relevant data from medical literature. METHODS: Analysis of the relation between serum lipid profile and postmenopausal changes, evaluation of positive and negative effects of estrogens on vascular wall and lipid metabolism, analysis of methods for the assessment of cardiovascular risk and evaluation of recent guidelines. RESULTS: Postmenopause is connected with significant changes in lipid metabolism, serum lipid profile and with increased risk of cardiovascular diseases. Deficit of estrogens influences lipid metabolism negatively. However, estrogen substitution has both positive and negative effects on vascular wall. Negative effects are: increased occurence of postprandial hyperlipidemia with increased triglycerides, generation of aterogenous small dense LDL particles, increased risk of inflammatory changes in vascular wall and procoagulation situation. CONCLUSION: Hormonal therapy can display some positive effects of vascular wall. However, recent data evaluate hormonal substitution with regard to atherosclerosis and cardiovascular problems as less benefitial or even risky.

[Serum amyloid A as an effective marker for the assessment of surgical trauma and risk of post-operative complications]. / Sérový amyloid A jako efektivní marker stupne operacní záteze a rizika komplikací.

OBJECTIVE: To analyze relations among acute phase reactants in a group of 40 women operated for uterine myom by laparoscopy and open surgery. DESIGN: Prospective study. METHODS: Plasma concentrations of C-reactive protein (CRP), serum amyloid A (SAA) and interleukin 6 (IL-6) were measured together with leukocytes in blood before operation, 24 and 72 hours post operation, respectively. RESULTS: Leukocytes and IL-6 displayed minimal response and decreased quickly after operation to preoperative levels. Concentrations of CRP and SAA remained increased after operation. There were no relationships between leukocytes and acute phase reactants. Normal leukocytes 72 hours post operation were found in 1/3 of women with increased at least one acute phase reactants and in 1/4 of women with increased at least two markers. Typ of surgery, surgical stress and length of surgery were related to the concentration of CRP, IL-6 and SAA. CONCLUSION: Changes in SAA 24 hours after operation are similar to CRP and IL-6. Surgical stress, length of operation and possible risk 72 hours after operation are best predicted by CRP and SAA (at that time IL-6 and leukocytes are practically normal). Maximal increase was found for SAA concentrations. Thus SAA seems to be suitable marker of early postoperative complications.

Attenuation and immunogenicity of recombinant yellow fever 17D-dengue type 2 virus for rhesus monkeys.

A chimeric yellow fever (YF)-dengue serotype 2 (dengue 2) virus was constructed by replacing the premembrane and envelope genes of the YF 17D virus with those from dengue 2 virus strains of Southeast Asian genotype. The virus grew to high titers in Vero cells and, after passage 2, was used for immunogenicity and attenuation studies in rhesus monkeys. Subcutaneous immunization of naive rhesus monkeys with the 17D-D2 chimeric virus induced a neutralizing antibody response associated with the protection of 6 of 7 monkeys against viremia by wild-type dengue 2 virus. Neutralizing antibody titers to dengue 2 were significantly lower in YF-immune animals than in YF-naive monkeys and protection against challenge with wild-type dengue 2 virus was observed in only 2 of 11 YF-immune monkeys. An anamnestic response to dengue 2, indicated by a sharp increase of neutralizing antibody titers, was observed in the majority of the monkeys after challenge with wild-type virus. Virus attenuation was demonstrated using the standard monkey neurovirulence test. The 17D-D2 chimera caused significantly fewer histological lesions than the YF 17DD virus. The attenuated phenotype could also be inferred from the limited viremias compared to the YF 17DD vaccine. Overall, these results provide further support for the use of chimeric viruses for the development of a new live tetravalent dengue vaccine.

The importance of age and statin therapy in the interpretation of Lp-PLA(2) in ACS patients, and relation to CRP.

C-reactive protein (CRP) is a marker of arterial inflammation while lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is related to plaque instability. The aim of this study was to evaluate the correlation between the risk of unstable plaque presenting as acute coronary syndrome (ACS) and Lp-PLA(2), and to assess the influence of statins on interpretation of Lp-PLA(2). A total of 362 consecutive patients presenting to the emergency department (ED) with acute chest pain suggestive of ACS were evaluated by cardiologists as STEMI, NSTEMI, or unstable angina, and non-ACS. Serum biomarkers measured on admission: troponin I, C-reactive protein (Abbott), and Lp-PLA(2) (DiaDexus). Four groups were defined according to the final diagnosis and history of statin medication: ACS/statin-; ACS/statin+; non-ACS/statin-; non-ACS/statin+. Lp-PLA(2) was highest in ACS/statin- group; statins decreased Lp-PLA(2) both in ACS and non-ACS of about 20 %. Lp-PLA(2) was higher in ACS patients in comparison with non-ACS patients group without respect to statin therapy (p<0.001). Lp-PLA(2) predicted worse outcome (in terms of acute coronary syndrome) effectively in patients up to 62 years; limited prediction was found in older patients. C-reactive protein (CRP) failed to discriminate four groups of patients. Statin therapy and age should be taken into consideration while interpreting Lp-PLA(2) concentrations and lower cut-off values should be used for statin-treated persons.

Construction, characterization and immunogenicity of recombinant yellow fever 17D-dengue type 2 viruses.

Chimeric yellow fever (YF)-dengue type 2 (Den 2) viruses were constructed by replacing the premembrane (prM) and envelope (E) genes of YF 17D virus with those from Den 2 virus strains of south-east Asian genotype. Whereas viable chimeric viruses were successfully recovered when the YF 17D C gene and the Den 2 prM gene were fused at the signalase cleavage site, no virus could be rescued from the constructions fused at the viral protease cleavage site. Unlike YF virus that replicated in all the cell lines tested and similar to the Den 2 virus, the recombinant viruses did not replicate in vaccine-production certified CEF and MRC5 cells. Besides, chimeric 17D/Den 2 viruses and their parental viruses reached similar growth titers in Vero and C6/36 cell cultures. Analysis of mouse neurovirulence, performed by intracerebral inoculation, demonstrated that the 17D/Den 2 chimera is more attenuated in this system than the YF 17DD virus. Immunization of mice with this chimera induced a neutralizing antibody response associated with a partial protection against an otherwise lethal dose of mouse neurovirulent Den 2 NGC virus. Overall, these results provide further support for the use of chimeric viruses as an attractive methodology for the development of new live flavivirus vaccines.

Introduction of a new laboratory test: an econometric approach with the use of neural network analysis.

We designed a simulation model for the assessment of the financial risks involved when a new diagnostic test is introduced in the laboratory. The model is based on a neural network consisting of ten neurons and assumes that input entities can have assigned appropriate uncertainty. Simulations are done on a 1-day interval basis. Risk analysis completes the model and the financial effects are evaluated for a selected time period. The basic output of the simulation consists of total expenses and income during the simulation time, net present value of the project at the end of simulation, total number of control samples during simulation, total number of patients evaluated and total number of used kits.

A strategy to promote the rational use of laboratory tests. International Federation of Clinical Chemistry.

These guidelines suggest how clinical chemists may promote rational laboratory use by critically evaluating the tests they offer. This may be done by documenting the clinical uses and limitations of tests in the same way that properly written up method descriptions outline analytical procedures in a particular laboratory. This information can then be promoted on result report forms, in discussion or through investigational protocols or clinical guidelines. For these to be of value they should be the result of clinical and laboratory input reflecting the local practice and facilities. They should be clearly written, easily accessible to the users and updated whenever there is a change of methodology or clinical practice. Much can be achieved by applying common sense to common knowledge and laboratories of all sizes can prepare their own material. However, national scientific societies and other professional organisations can assist local endeavour by avoiding and disseminating educational material on test evaluation, selection and use.

Midterm follow-up study of laparoscopic dissection of uterine vessels for surgical treatment of symptomatic fibroids.

BACKGROUND: This study aimed to assess laparoscopic dissection of uterine vessels (LDUV) for symptomatic fibroids in women. METHODS: A total of 69 women entered the study between March 2000 and June 2003. In this case series, 68 consecutive women underwent LDUV using ultrasonically activated sheers or electrosurgery for the treatment of fibroids over 3 years (median follow-up period, 14.5 months). Ultrasound or magnetic resonance imaging was carried out 3, 6, 12, 24, and 36 months after treatment. The tissue markers, gonadotropin, and estrogen levels were studied postoperatively. RESULTS: Almost all the patients (98.5%) had a successful LDUV with a low rate (7.3%) of postoperative complications. The time of surgery ranged from 15 to 50 min (mean, 30.8 min). The blood loss was minimal (mean, 14.7 ml), and the hospital stay was 2.4 days. Symptom improvement (menorrhagia or dysmenorrhoea) was 93.2%, and the average reduction in the dominant myoma was 57.8% during a follow-up period longer than 12 months. All the patients with anemia had normal red cell counts after 3 months. CONCLUSIONS: Uterine volume and the dominant fibroid were significantly reduced and symptoms were improved by LDUV. The laparoscopic procedure is associated with insignificant tissue damage and normal gonadotropin and estrogen levels.

The yellow fever 17D vaccine virus: molecular basis of viral attenuation and its use as an expression vector.

The yellow fever (YF) virus is the prototype flavivirus. The use of molecular techniques has unraveled the basic mechanisms of viral genome structure and expression. Recent trends in flavivirus research include the use of infectious clone technology with which it is possible to recover virus from cloned cDNA. Using this technique, mutations can be introduced at any point of the viral genome and their resulting effect on virus phenotype can be assessed. This approach has opened new possibilities to study several biological viral features with special emphasis on the issue of virulence/attenuation of the YF virus. The feasibility of using YF virus 17D vaccine strain, for which infectious cDNA is available, as a vector for the expression of heterologous antigens is reviewed.

Comparison of two procedures for laparovaginal hysterectomy: a randomized trial.

OBJECTIVE: To compare peroperative parameters of two variants of a laparovaginal hysterectomy in surgical management of gynecological conditions. METHODS: A prospective randomized study of 70 laparovaginal hysterectomies performed by the same two surgeons for disease of female pelvic organs. The following criteria were studied: indication for surgery, previous surgery, duration of the procedure, recovery, hospital stay, blood loss, tissue damage markers, hysterectomy proportions and complication incidence. Statistical analysis was performed using the non-parametric chi(2)-test and non-parametric Fischer's exact probability test when appropriate, with a level of significance P=0.05. RESULTS: Totals of 38 (54.3%) laparoscopy-assisted vaginal and 31 (45.7%) vaginally assisted laparoscopic hysterectomies were performed for fibroma as the main indication. Conversion to laparotomy was applied in only one patient. The VALH group (P=0.01) showed both fewer procedures and shorter hospital stay with insignificant blood loss. CONCLUSION: The two variants of a laparovaginal hysterectomy appear to be safe and appropriate, effective procedures for women with gynecological conditions. Furthermore, vaginally assisted laparoscopic hysterectomy has been shown to be superior to laparoscopy-assisted vaginal hysterectomy in terms of shorter operating time and greater palliative effect upon the complex of uterosacral ligaments. Laparoscopic surgery can alter the relationship between vaginal and abdominal hysterectomy.

Laparoscopic hysterectomy in obese women: a clinical prospective study.

OBJECTIVE: To compare perioperative and postoperative outcomes of laparoscopic hysterectomy (LH) in surgical management of gynecological conditions in two groups of different weight. METHODS: A prospective comparative clinical study of 271 LH performed for disease of female pelvic organs in a group of 54 obese patients (over 30 body mass index (BMI)) and in a group of 217 non-obese patients (less than 30 BMI). The following criteria were assessed: patient characteristics, indications for surgery, previous surgery, presence of adhesions, duration of procedure, blood loss, weight of specimen, hospital stay and complications. Statistical analysis was performed using the unpaired t-test and non-parametric Chi-square test when appropriate, with a significance level of P=0.05. RESULTS: Three non-obese patients were converted to laparotomy due to operative complications. Laparoscopy in the remaining 268 patients (98.89%) was completed successfully. There was no significant difference in estimated blood loss, presence and degree of adhesions, weight of specimen, length of hospital stay and postoperative complications between women with high BMI and those with low BMI. The rate of major operative complications (5.55% versus 3.22%) was higher in the obese group. The duration of the operation was longer in obese women. However, the significance of the difference was borderline (P=0.06).

Laparoscopic staging of endometrial cancer using laparosonic instruments: comparison with electrosurgery.

OBJECTIVE: To compare perioperative parameters in two groups of women with different laparoscopic operative techniques in surgical staging of endometrial cancer (EC). STUDY DESIGN: Thirty randomly allocated and laparoscopically treated women with EC. Fifteen patients were operated by electrosurgery, 15 patients by laparosonic operative technique. Differences between the two groups were determined by the Wilcoxon rank-sum test. Probability (P) of less than 0.05 was considered significant. SETTING: Department of Gynecology and Obstetrics, Endoscopic Training Center, Baby Friendly Hospital, Kladno, Czech Republic. RESULTS: Laparoscopy was successfully completed in 29 patients. Laparoscopy-assisted surgical staging of EC was performed based on the tumor grade and the depth of myoinvasion. In both groups, in total 18 and 5 women underwent pelvic lymphadenectomy (PLN) and infra-aortic lymph node sampling (IALS), respectively. Three patients had metastases in pelvic lymph nodes. In the electrosurgical hemostasis and laparosonic group the mean total time required to finish the whole operative procedure were 132.1 and 138.3 min, respectively, with no statistically significant difference (P=0.96). There were no significant differences between the groups in any intraoperative or postoperative follow-up variables, except for the number of excised lymph nodes where the difference between electrosurgery and laparosonic group (12.7 versus 18) was statistically significant (P=0.05). In one patient with intraoperative venous bleeding the laparosonic hemostasis was ineffective (successful procedure rate 93.3%). One patient from the electrosurgery group was converted to laparotomy due to injury to the epigastric vessels. This complication had no connection with the surgical techniques studied. CONCLUSION: It is concluded that both operative technique variants in laparoscopy-assisted surgical staging appear to be feasible and effective for patients with EC.

Laparoscopic lymph node dissection using ultrasonically activated shears: comparison with electrosurgery.

PURPOSE: To assess and compare perioperative parameters in two groups of patients treated by different laparoscopic techniques of lymph node dissection (LND) for gynecologic cancer. PATIENTS AND METHODS: Between April 1996 and March 2001, 59 consecutive women with microinvasive cervical cancer (N = 5) or clinical stage I endometrial cancer (N = 54) underwent laparoscopic LND during a primary staging procedure using an electrosurgery (ELC) or ultrasonic (US) operative technique. The two groups were compared for perioperative outcomes. Differences between the two groups were determined by the Wilcoxon's rank-sum test. RESULTS: Laparoscopic LND and other staging procedures were completed successfully in 58 women (98.3%). There were no statistically significant differences between the groups with regard to perioperative outcomes (operation time, time for LND, blood loss, hospital stay, complications), but there was a significant difference (P = 0.0008) in the number of lymph nodes harvested: a mean of 13.7 in the ELC group and 17.5 in the US group. The pathologists found that the reading of histology slides was easier after US dissections because of the greater depth of thermal injury in the lymphatic tissue in ELC group. CONCLUSION: The US operative technique ensures efficient coagulation, cutting, dissection, and grasping for laparoscopic LND in patients with cervical and endometrial cancer.

Comparison of two procedures for sentinel lymph node detection in patients with endometrial cancer: a pilot study.

OBJECTIVE: The purpose of this study was to assess the feasibility and contribution of two intraoperative procedures of lymphatic mapping and sentinel node detection using a blue dye in surgically-staged patients with early stage endometrial cancer. METHODS AND MATERIALS: In 25 cases of endometrial cancer, patent blue-V was injected into the subserosal myometrium (13 cases, SM group) or cervico-subserosal myometrium (12 cases, CSM group) during a surgical staging procedure. Laparoscopically-assisted vaginal hysterectomy and pelvic lymphadenectomy were completed successfully in 23 women out of 24 laparoscopically-staged patients (95.8%). One patient with FIGO stage IIa was indicated for a radical abdominal surgery. RESULTS: A deposition of the blue dye was found in at least one pelvic lymph node (LN) in eight out of 13 cases (61.5%) in the SM group compared with ten out of 12 cases (83.3%) in the CSM group (p = 0.378). The mean number of dye-colored LN (DCLN) was 1.15 (SM group) and 2.5 (CSM group), respectively (p = 0.05). The rate of DCLN/LN was 15/188 (SM group) versus 30/190. respectively (p = 0.03). An uptake of the blue bye was observed in a total of 45 out of 388 LN. CONCLUSION: An intraoperative combination of cervico-subserosal myometrium application of the blue dye allows successful detection (83.3%) of sentinel LN in patients with endometrial cancer. Comparing SM and CSM groups the statistical significant difference was found in the DCLN/LN rate and mean number of sentinel lymph nodes (p = 0.03, p = 0.05, respectively). Clinical validity of this surgical procedure must be assessed prospectively.