Simultaneous infection with gammaherpes and influenza viruses enhances the host immune defense.

We have studied the impact of simultaneous infection of mice with murine gammaherpesvirus (MHV) and influenza A virus (IAV) on the immune response and pathogenesis of both infections. After a persistent MHV-68 herpesviral infection had been established, the same mice were super-infected with IAV. Individual parameters of MHV infection (viral DNA detection in organs and blood) and numbers of leukocytes in lungs and spleens were determined. With regard to the assumed reactivation of MHV-68 (mainly in lungs, spleen, thymus and peritoneal exudate cells) we focused our attention on the detection of transcripts, typical either for lytic infection (ORF50) and/or for latency (ORF73). Herpesviral DNA was detected in above mentioned organs in several intervals during the acute phase of IAV co-infection, but the expression of monitored transcripts was lower, i.e. it has decreased. Though the reason for such limited expression during acute influenza superinfection remains unclear, it is unambiguous that lower MHV-68 expression was detected in lungs and peritoneal exudate cells (PECs) from 3rd to 10th day after co-infection with IAV. Furthermore, our study showed that the ongoing gammaherpesvirus latency in co-infected mice affected the number of cytotoxic T-lymphocytes and neutrophils during the acute IAV infection and lowered their deviations from that of non-infected mice. Therefore, we suppose that co-infection with herpes and influenza viruses could be mutually beneficial for the host by promoting its defense against both viruses.

Possible role of different animal species in maintenance and spread of murine gammaherpesvirus 68 in the nature.

Murine gammaherpesvirus 68 (MHV-68), isolated from a bank vole (Clethrionomys glareolus) in Slovakia in 1976 is a natural pathogen of wild murid rodents. This review is focused to biological properties of this pathogen, the mode of its maintenance in murid rodents as reservoir animals, mechanisms of its spread to other animals in the same biotope as well as to livestock and household animals. Potential role of ticks as vectors and the possibility of infection of humans with this virus are considered as well. All the above evidence of the virus infection of various hosts is based on serological or molecular analytical data. The presented knowledge indicates important epizootologic consequences, namely harboring and permanent maintenance of the virus in murid rodents as reservoir animals with a real possibility of spread to other animals in the same biotope. These relationships imply a cross-species virus transmission with potential serious consequences for the infected animals or humans.

Biological and pathogenetic characterization of different isolates of murine gammaherpesvirus 68 (MHV-68) in the context of study of human oncogenic gammaherpesviruses.

Study of murine gammaherpesvirus 68 (MHV-68), which was discovered in 1980 in Slovakia, has led to many important findings regarding gammaherpesviral properties in general. Nowadays, it is considered to be a universal model used for detailed studies to determine pathogenetic, immunological and molecular aspects of oncogenesis in analogy to Epstein-Barr virus (EBV) and Kaposi΄s sarcoma-associated virus (KSHV). The objective of this work is to characterize biological and pathogenetic properties of the virus with an emphasis on our prior results concerning ecology, epidemiology, viral persistence in peritoneal macrophages, detection of malign and benign lymphoproliferations accompanied by the presence of atypical lymphocytes in blood during IM-like and leukemia-like syndromes. We are trying to elucidate the role of virus-specific genes in virulence, pathogenicity and murine gammaherpesvirus oncogenesis by comparison of molecular-biological, pathogenetic and oncogenic potential of MHV-68 isolates and deletion mutant MHV-76 and therefore help to understand the analogical processes that occur in EBV infected patients.

Vertical transmission of murine gammaherpesvirus 68 in mice.

The mouse infected with murine gammaherpesvirus 68 (MHV-68) is accepted animal model for investigation of pathogenesis, oncogenesis, immunology and molecular biology of gammaherpesviruses in their natural host. However, little is known about the host range, epidemiology and pathogenesis of this natural pathogen of free-living murid rodents. Therefore we addressed the question of transplacental transmission of MHV-68 from pregnant Balb/c mice chronically infected with the virus to their fetuses and shedding of the virus by breast milk from chronically infected mothers to their offspring. The mothers were positive to infectious virus and viral antigen in various organs but mainly in the spleen and peritoneal *macrophages. Virus-neutralizing serum antibodies, and leukocyte count with a proportion of atypical lymphocytes were increasing to elevated values with the time of infection. The chronic infection resulted in premature termination of pregnancy or reduced number and size of newborns due to their retarded development. Out of 10 infected pregnant mice just one died and another developed a tumor. All these results confirm the vertical transmission of MHV-68 in mice, teratogenicity of the virus and the virus shedding by breast milk.

MDR1 (C3435T) polymorphism: relation to the risk of breast cancer and therapeutic outcome.

P-glycoprotein (PGP), the product of the MDR1 gene, is a transmembrane active efflux pump for a variety of carcinogens and cytostatics. It has been suggested that MDR1 polymorphisms contribute to the variability in cancer risk and therapeutic outcome. We examined the relevance of C3435T polymorphism in relation to breast cancer susceptibility, clinical and pathological characteristics of breast carcinoma, the therapeutic response and hematologic toxicities after anthracycline-based chemotherapy. A significant association between allele frequencies and histological type, stage and histological grade was observed (P=0.024, 0.014, 0.006, respectively, chi(2)-test or Fisher's exact test). We also found significantly higher (P=0.019, chi(2)-test) T allele frequency in breast cancer patients (n=221) than in controls (n=113). A significantly enhanced therapeutic outcome after neoadjuvant therapy (n=38; P=0.021, Fisher's exact test) and longer time to progression after anthracycline-based chemotherapy (n=102; P=0.049, log-rank test) were observed in CC homozygotes. However, no significant association between hematologic toxicities and C3435T polymorphism was detectable.

Expression of MDR proteins in breast cancer and its correlation with some clinical and pathological parameters.

The aim of this work was to determine the expression of the multidrug resistance (MDR) proteins, namely MDR1 (P-glycoprotein), MRP1 (multidrug resistance-related protein) and LRP (lung resistance-related protein), in 87 samples of breast carcinoma. Detection of these proteins was provided by using indirect enzymatic immunohistochemistry. Our findings were compared with the other clinical and pathological parameters: expression of Her2/neu, estrogen receptor status (ER), progesteron receptor status (PR), histological grade and regional lymph node status. For statistical analysis, non-parametric two sided Mann-Whitney-U test was used. Majority of breast carcinoma specimens show positivity for these proteins. The MDR1 and MRP1 signal was found in the cytoplasm of cancer cells. The expression of LRP was detected in the cytoplasm close to the nuclear membrane. The samples were positive for MDR1 protein in 57%, for MRP1 in 84% and for LRP in 79%. Comparing our results with other clinical and pathological parameters, negative correlation between ER, PR and MDR1 expressions and histological grading status was found. No associations were observed between the MRP1 and LRP proteins and histological grading, as well as between the expression of three MDR proteins and the other clinically relevant parameters. In conclusion, high frequency of expression of MDR proteins in breast carcinoma cells suggests, that these proteins might be an important factor of drug resistance in breast carcinoma. Nevertheless, the negative correlation between the histological grade of malignancy of tumor and the expression of ER, PR and MDR1 indicates possible influence of progressive tumor cell de-differentiation. However, this finding has to be confirmed in additional evaluations.

[Endometrial thickness in postmenopausal patients with breast carcinoma--preliminary results]. / Hrúbka endometria u postmenopauzálnych pacientok s karcinómom prsníka--predbezné výsledky.

OBJECTIVE: One of the most important factors contributing to developing of the breast cancer is a female organism exposure to estrogens. Thickness and structure of endometrium in postmenopausal women is a sign of the hormonal status. The goal of the study was to investigate whether there is a difference in the endometrial thickness and quality in postmenopausal women with breast cancer. DESIGN: Prospective cohort study. SETTING: Ultrasound Unit, 2nd Department of Obstetrics and Gynecology, Medical School and Medical Teaching Hospital of L. Pasteur, Kosice, Slovakia. MATERIAL AND METHODS: Endometrial thickness was measured by transvaginal ultrasonography in 21 patients with histologically verified breast cancer and was compared with a group of 26 well women. We assessed the thickness of endometrium, the size of uterus and the endometrium/myometrium index. RESULTS: Despite the fact there existed a difference between these two groups (the average thickness of 3.6 mm and 2.9 mm), the result was not statistically applicable to show a correlation with a benign or malignant breast tissue. CONCLUSION: The results may support the theory of breast cancerogenesis in correlation with estrogen influence or increased incidence of estrogen receptors, respectively. As definitive statistical result can not be stated due to a limited group of patients, more investigation might be necessary in the future. The study dealing with a similar problem was not found in available publications.

Vinorelbine, epirubicin, and methotrexate (VEM) as primary treatment in locally advanced breast cancer.

PURPOSE: This phase II trial of VEM (vinorelbine + epirubicine + methotrexate) in the treatment of locally advanced breast cancer was conducted to obtain downstaging to allow surgery and breast conservation. PATIENTS AND METHODS: This multicenter study recruited 58 patients with locally advanced breast cancer (two patients ineligible); 56 were evaluable for response and tolerance. RESULTS: Downstaging was obtained in 77% of the patients with a pathological complete response (pCR) rate of 9%. At 33 months of follow-up, median survival has not been reached. Neutropenia grade 3-4 was reported in 31% of cycles with 3% of cycles with infection grade 3. Alopecia grade 3 was noticed for 71% of patients. CONCLUSION: VEM represents an effective regimen for patients with locally advanced breast cancer, allowing an important pCR. Moreover, this regimen appears to be particularly well tolerated.

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).

Does hysterectomy without salpingo-oophorectomy influence the reoperation rate for adnexal pathology? A retrospective study.

OBJECTIVE: The aim of this study was to assess if the abdominal, vaginal and laparoscopic approach to hysterectomy can affect the incidence of ovarian or adnexal pathology after hysterectomy without salpingo-oophorectomy. METHODS: In this study 17 cases out of 617 hysterectomies were found to have development of adnexal pathology; reoperation rate was 2.75%. RESULTS: The reoperation rate was significantly different (p < 0.006) in the observed approaches to hysterectomy (TAH 5.67%, VH 0.69%, LH 3.18%). The greatest difference was found between the abdominal and vaginal groups. CONCLUSION: Our study results have sufficiently shown that the relationship of a number of factors (age, primary histologic findings, smaller peritoneal trauma) had an important impact on a significant difference in reoperation rate between vaginal, laparoscopic and namely abdominal hysterectomies in female patients with preserved adnexa.

[Hormone replacement therapy in women with surgical treatment of endometriosis and adenomyosis: prospective and follow-up study. Part I]. / Hormonální substitucní lécba u zen operovaných pro endometriózu a adenomyózu: prospektivní follow-up studie--I. Cást.

OBJECTIVE: To determine whether the initiation of estrogen replacement therapy (ERT) in the postoperative period increases the incidence of symptom recurrence following laparoscopic hysterectomy (LH) and/without bilateral salpingo-oophorectomy (BSO) for the treatment of endometriosis and adenomyosis. DESIGN: Prospective partially randomized patient preference follow up trial (PRPPT). SETTING: Department of Gynaecology, Endoscopic Training Centre, Kladno Hospital. METHODS: Chart review of 286 consecutive patients who underwent LH and/or without BSO for endometriosis or adenomyosis between April 1994 and June 1999 and who subsequently received ERT. The methods of second phase of trial was prepared. The symptoms of recurrence in patients who started ERT after surgery and in those who did not start ERT (control group) will be compared and adjusted. RESULTS: Seventy-three percent had preoperative pelvic pain. One hundred thirty two women (46.2%) had only endometriosis. Of the cases of endometriosis, 35.4% was stage I and 64.4% was stage II.-IV. One hundred fifty four women had adenomyosis, 60 with endometriosis. CONCLUSION: In the first phase of follow study the collection of clinical data was performed. Preliminary results from the PRPPT demonstrate that laparoscopic approach to surgery for endometriosis increased significantly the number of this disease.

An epidemiologic-immunological inquiry into the incidence of altered immunity in children of school age.

The authors examined 536 children 10 years of age to evaluate their immune status and the development of allergies and recurrent respiratory inflammations. Altered resistance was found in as many as 47% of children, pollen allergy and atopic eczema in at least 10.5%, habitual rhinitis in 15.2%, asthma in 3.5% and recurrent inflammatory infections in 13.1%. A higher rate of alterations was found where environmental conditions were deteriorated. Elevated IgE, changes in IgG, IgA and IgM as well as increased levels of acute phase proteins suggest a relationship between the degree of immune impairment and clinical manifestations. Our results imply there are large numbers of children with altered immunity and point to the fact that in common practice only 14% of them are attended to by the clinical immunologist and thus receive causal treatment. It is therefore concluded that, in line with the expansion of noxious factors which accompany our civilization, more attention should be given to the development of immunity in the young population.

Immunoglobulins and some serum proteins in children with altered resistance coming from areas with variously polluted atmosphere.

In four areas with different types of atmospheric pollution 534 children of school age were examined for serum immunoglobulins (IgG, IgA, IgM and IgE), saliva IgA (sIgA), lysozymes (LYS) and acute phase reactants (alfa-l-antitrypsin - A1AT, alfa-2-macroglobulin -A2M, ceruloplasmin -CPL, transferrin - TRF). The children were divided into two groups: those with altered resistance (AR) comprising allergoses of all types and recurrent or persistent infections and those without the above health problems, i.e. healthy (H). There were more children with AR in areas with higher atmospheric pollution than in control areas. The frequency of AR was higher among boys than among girls. In the control area BN, a number of parameters in the AR group differed significantly from those in the H group. In areas with substantial atmospheric pollution these differences were generally less pronounced. Significant differences were found between the control and polluted areas in many indicators. In the area KO characterized by an intermediate degree of industrial pollution the means of the tested parameters were habitually elevated, whereas in the heavily polluted areas they were decreased. The most sensitive tests for evaluating differences between the areas were the levels of A1AT, LYS, and IgE, while the variations of sIgA, sLYS and CPL were less pronounced.

Immunological profiles in workers occupationally exposed to inorganic mercury.

Forty-one male mercury-exposed workers were examined for the serum concentration levels of immunoglobulins (IgG, IgA, IgM), alpha-1-antitrypsin (A1AT), alpha-2-macroglobulin (A2M), ceruloplasmin (CPL) and orosomucoid (ORO). In the period preceding this investigation the mercury concentrations in workplace air ranged from 0.106 to 0.783 mg.m-3, the range of urinary mercury concentrations was from 0.029 to 0.545 mg.1(-1). All but two (IgG and A1AT) of the immune parameters tested were at the levels that were evidently higher than those found in a control group of 55 workers matched by age who lived in a relatively clean area. The percentage of individuals with no value out of the range of normal physiological limits was in the controls almost 80%, in the exposed 36.6% only. These findings confirm the literature data which show that an inhalation exposure to inorganic mercury may evidently have a stimulating effect on some serum proteins in humans.

The immune profile of contact allergy patients.

Nonspecific parameters of antibody-related and cellular immune responses were studied in a group of 30 patients treated for manifest contact allergy to metals (chromium, cobalt, nickel) or non-metals (epoxy resin, rubber) confirmed in each of them by specific patch test positivity. In addition, every patient was tested for the pattern of skin reactivity to the Immunoskintest (USOL, Prague) antigens administered intradermally. The group of 20 metal allergy patients had decreased suppressor index values, enhanced formation of immediate active rosettes and an elevation of serum IgM and IgA immunoglobulins. The remaining 10 patients reacted by a reduced ability of neutrophils to exert phagocytic action on HEMA particles and a decrease in levels of alpha-2-macroglobulins in serum. This may help select a series of suitable immune parameters to be used as prediction tests in the secondary prevention of cases of occupational contact dermatitis.

Lack of correlation between serum and salivary concentration levels of immunoglobulin A and lysozyme (muramidase).

The study was conducted in a group of 232 individuals comprising 114 males and 118 females 20 to 67 years old. As revealed by the linear regression analysis method, mean serum concentration levels of immunoglobulin A (IgA) and lysozyme (LYS) were unrelated to values found in the spontaneously secreted saliva specimens. Levels of serum IgA were clearly age-dependent, but no such correlation was recorded in the case of secretory IgA. These findings suggest that tests for salivary IgA can hardly be taken as an equivalent to IgA determinations in the human serum.

Relationship of blood protein levels to outdoor air pollutant concentrations in a semicohort of school-age children living in urban areas differing by quality of air.

Variations in some humoral immune responses to polluted air were studied in two semicohorts of children, initial age 10 years, from two urban communities differing from each other by the degree of ambient air pollution. The material for analysis (blood, saliva) was collected every autumn and spring in 3 successive years, giving a total of 6 sets of specimens for each examinee. All blood specimens were examined for the serum level of immunoglobulins (IgG, IgA, IgM), lysozyme (LYS), total serum protein (TP) and the level of the acute reactants alpha 2 macroglobulin (A2M), alpha 1 antitrypsin (A1AT), transferrin (TRF) and ceruloplasmin (CPL). The saliva specimens were examined for the level of lysozyme (sLYS) and secretory IgA (sIgA). The mean protein concentrations for each of the 6 sampling series were correlated with the mean of 24-h emission concentrations measured in the last 3 months preceding the autumn or spring sampling series. In the community area characterized by a low-degree non-industrial pollution of air the correlations of immunoglobulins to SO2 and floating particles (FP) in air were as a rule inversed while the response from TP, LYS and acute reactants was direct. In the community contaminated by industrial pollutants, correlations between proteins and SO2 were markedly weaker, but there was a significant positive correlation between H2S and levels of IgA and A2M in blood and sIgA and sLYS in the saliva. A high degree of positive correlation was also observed between H2S and levels of IgM and LYS. Inverse correlations were only between levels of LYS and FP, SO2 and H2S. Significant correlations were also between contaminant concentrations and FP. The associations found between the contaminant concentrations in air and levels of blood and saliva proteins supports the hypothesis that quality of air may have considerable impacts on defense mechanisms. Seasonal variations in the quality of air may increase the rates of childhood morbidity for acute upper respiratory tract infections.

Immunological profiles in workers of a power plant burning coal rich in arsenic content.

A group of 47 male adults working in a thermal power plant burning coal containing 900 to 1,500 g of arsenic per ton dry weight was examined on the blood serum immunoglobulins IgG, IgA and IgM content and levels of acute reactants alpha-1-antitrypsin (A1AT), alpha-2-macroglobulin (A2M), transferrin (TRF), orosomucoid (ORO) ceruloplasmin (CPL), and lysozyme (LYS). Investigations in the control group comprising 27 workers from another power plant in the same district where the coal content of arsenic was more than 10 times lower were analogous. The inter-group differences in means were evaluated by t-test, differences in the association of values by F-test, and the correlations with age and the length of exposure were assessed using the regression analysis method. The differences in mean IgG, IgA, IgM, LYS and A2M levels between the exposed and control groups of workers were insignificant or of borderline significance only. In contrast, differences in TRF, ORO and particularly CPL levels were statistically highly significant, in all instances P less than 0.001. In the control group, persons with abnormal values in at least two immunobiochemical tests used accounted for 3.7%, in the group of the exposed for 51% (P less than 0.002). All these findings, especially the rise in CPL concentration levels in the exposed group are discussed on the background of the rise in cancer mortality rates found previously in this group of power plant workers.

Immunobiochemical profiles of workers differing in the degree of occupational exposure to vinyl chloride.

The immunobiochemical studies were conducted in a group of 98 production workers engaged in polyvinyl chloride manufacture from ethylene (group A workers) and in a group of 59 vinyl chloride workers from a chemical plant employing classic production technology from acetylene (group B workers). Both groups of workers were matched by age (group A workers: 37.7 +/- 8.66 years; group B workers: 34.9 +/- 11.2 years) and average exposure length (group A workers: 8.6 +/- 3.0 years; group B workers: 10.7 +/- 8.4 years). All workers were examined for the serum concentrations of immunoglobulins IgG, IgA and IgM and acute reactants lysozyme (LYS), transferrin (TRF), ceruloplasmin (CPL), alpha-l-antitrypsin (AlAT), alpha-2-macroglobulin (A2M) and orosomucoid (ORO). The statistical analysis included calculations of means, standard deviations and 95% confidence intervals. Differences in means were evaluated by t-test, differences in the distribution pattern of values by F-test. Abnormality of values was assessed by comparisons to normal values valid in Czechoslovakia. Group A worked in conditions meeting the MAC 10 mg VC.m-3 comparing with group B workers had elevated levels of IgG (P less than 0.005), IgA and IgM (P less than 0.001 both). Group B workers differed from group A workers by exhibiting significantly elevated levels of AlAT, and CPL. (P less than 0.001). The differences in the frequency of abnormal values between group A and group B worked in substantially less favourable hygienic conditions were significant for immunoglobulins elevated in group A and for ORO (P less than 0.01) and CPL (P less than 0.001) elevated in group B. The possible relationship of these immunobiochemical findings with the degree of vinyl chloride exposure are critically analyzed.

Factors of humoral resistance in workers exposed to vinyl chloride with a view to smoking habits.

Serum samples were assayed in 110 workers (59 smokers and 51 non-smokers) at PVC manufacturing factory, the results being compared with those obtained in a group of age-matched healthy controls. Non-smokers had significantly higher levels of immunoglobulins (IgG, IgA, IgM), while in smokers there was an increase in IgM only. Lysozyme levels (LYS) were elevated in all exposed subjects, but there was a highly significant decrease in the total protein (TP) content. Alfa-2-macroglobulin (A2M) and orosomucoid (ORO) were also elevated in exposed workers. A significant increase was found in ceruloplasmin (CPL), with smokers having higher levels than non-smokers. No difference was detected between the examined groups in transferrin (TRF) and alfa-1-antitrypsin (A1AT). Exposure duration did not correlate with any of the assayed parameters. The positive correlation of IgA levels and inverse correlation of CPL with age in the control group could not be confirmed in the exposed subjects where a significant inverse correlation between age and AlAT levels, which was found to be positive in exposed non-smokers, proved to be inverse in the subgroup of smokers. The authors discuss possible explanations of the reported findings.