Mixed infections with distinct cytomegalovirus glycoprotein B genotypes in Polish pregnant women, fetuses, and newborns.

The purpose of this investigation was to describe a distribution of cytomegalovirus (CMV) single and multiple genotypes among infected pregnant women, their fetuses, and newborns coming from Central Poland, as well as congenital cytomegaly outcome. The study involved 278 CMV-seropositive pregnant women, of whom 192 were tested for viral DNAemia. Human cytomegalovirus (HCMV) genotyping was performed for 18 of 34 pregnant women carrying the viral DNA and for 12 of their 15 offspring with confirmed HCMV infections. Anti-HCMV antibodies levels were assessed by chemiluminescence immunoassay (CLIA) and enzyme-linked fluorescence assay (ELFA) tests. Viral DNA loads and genotypes were determined by real-time polymerase chain reaction (PCR) assays for the UL55 gene. In the pregnant women, we identified HCMV gB1, gB2, gB3, and gB4 genotypes. Single gB2, gB3, or gB4 genotypes were observed in 14 (77.8 %) women, while multiple gB1-gB2 or gB2-gB3 genotypes were observed in four (22.2 %). Maternal HCMV genotypes determined the genotypes identified in their fetuses and newborns (p ≤ 0.050). Half of them were infected with single HCMV gB1, gB2, or gB3 genotypes and the other half with multiple gB1-gB2 or gB2-gB3 genotypes. Single and multiple genotypes were observed in both asymptomatic and symptomatic congenital cytomegaly, although no gB3 genotype was identified among asymptomatic cases. In Central Poland, infections with single and multiple HCMV strains occur in pregnant women, as well as in their fetuses and neonates, with both asymptomatic and symptomatic infections. HCMV infections identified in mothers seem to be associated with the viral genotypes in their children.

In situ RT-PCR can distinguish between productive and latent cytomegalovirus infection in the blood cells of bone marrow transplant recipients.

Thirty-four peripheral blood leukocyte samples from bone marrow transplant (BMT) recipients were examined for Human cytomegalovirus (HCMV) phosphoprotein 65 (pp65), DNA and late transcripts. Twenty seven samples were positive for pp65 in the cytoplasm by immunofluorescent assay (IFA). Viral DNA was confirmed in 26 samples by nested PCR (nPCR). Using in situ RT-PCR, viral late transcripts were found in 19 samples, positive also by IFA and nPCR; these samples were considered indicative of productive viral infection. Five samples, positive by nPCR but negative by IFA and in situ RT-PCR, were considered to represent latent viral infection. In 8 samples, positive by IFA and nPCR but negative by in situ RT-PCR, apparently phagocytosis of viral particles took place.

Antiviral activity of Flos verbasci infusion against influenza and Herpes simplex viruses.

The lyophilized infusion from flowers of Verbascum thapsiforme Schrad. (FVI) showed antiviral activity in in vitro studies against Fowl plague virus, several influenza A strains, influenza B strain as well as Herpes simplex virus. Influenza viruses titer decreased by 1-3 log units, while of H. simplex virus by 2.3 log. FVI has shown virucidal activity on H. simplex virus at 300 micrograms/ml, but did not inactivate influenza viruses. Phytochemical investigations of FVI have shown the presence of flavonoids, iridoids, phenolic acids, saponins, amino acids and free sugars.

[Cytomegalovirus infections in infants examined in the department of virology in 1988-1992]. / Zakazenia niemowlat wirusem cytomegalii na podstawie obserwacji prowadzonych w Zakladzie Wirusologii w latach 1988-1992.

Active cytomegalovirus (CMV) infections were found in 91 infants (11%) examined in the Department of Virology from 1988 to 1992. Serological methods (ELISA, CFT) and virus isolation from urine were used to diagnose CMV infection. Congenital or perinatal infections were recognized in 20 infants, while 10 infants were infected by blood transfusion. In most cases the route of CMV transmission was not recognized. The occurrence of clinical symptoms in CMV-infected children was also studied. In 80% of the infants the disease was characterized by hepatosplenomegaly or liver dysfunction. CMV pneumonia was found in 41% of the cases, and gastroenterocolitis in 36% of the subjects.

[Epstein-Barr virus infection in patients after bone marrow and heart transplantation]. / Zakazenie wirusem Epsteina-Barr u pacjentów po przeszczepie szpiku kostnego i serca.

UNLABELLED: Epstein-Barr virus (EBV) infections in immunosuppressed patients cause the severe clinical problems. Posttransplant lymphoproliferative disease (PTLD) might occur as a result of the latent EBV activation. OBJECTIVE: Occurrence of active EBV infection in heart and bone marrow transplant patients. METHODS: 68 serum samples obtained from 13 allogenic bone marrow and 20 heart transplant patients were tested by IF and ELISA methods. Antibodies against VCA, EA and EBNA antigens were measured. RESULTS: All patients showed the presence of anti-VCA IgG antibodies, thus all were seropositive. Three patients (9%) showed primary EBV infection while in 12 (36%) patients virus reactivation or reinfection was confirmed. CONCLUSIONS: 1. EBV infection in immunosuppressed patients is mainly caused by latent virus reactivation. 2. Type of EBV infection can be confirmed serologically only by the detection of specific anti-VCA, EA and EBNA antibodies. 3. The risk of PTLD in transplant patients creates the need for frequent monitoring.

[Congenital and acquired cytomegalovirus infection in infants confirmed by virologic studies]. / Cytomegalia wrodzona i nabyta niemowlat potwierdzona badaniami wirusologicznymi.

Sixty infants in whom clinical symptoms suspected of cytomegalovirus (CMV) infection were studied. CMV infection was found in 50% of the subjects. The diagnosis was based on studies of specific antibodies and isolation of the virus from urine and/or throat swabs. In most of the children the examinations were repeated several times, and clinical observations continued for 1 to 42 months (avg. 18 months). IgM-class antibodies were detected in 26 children and in 18 the virus was isolated. In 3 infants, isolation of CMV virus was the only evidence of active infection. Persisting viruria (avg. 11 months) and long-term presence of Ig G antibodies, even to 44th month of life were also observed. Congenital infection was diagnosed in 4 infants; the remaining ones acquired the infection during the perinatal period or later. In 7 cases transfused blood cannot be excluded as the source of infection. The clinical symptoms manifested in infected and non-infected children were similar. There was a statistically significant increase in the occurrence of hepatomegaly, splenomegaly, hyperbiliru-binemia and diarrhoea in infected children. Congenital abnormalities were found in 10 infected children, including 4 cases of congenital cytomegaly.

[Factors influencing prevalence and clinical course of cytomegalovirus (CMV) infection in kidney transplant patients]. / Czynniki wplywajace na wystepowanie i przebieg kliniczny zakazenia wirusem cytomegalii (CMV) u chorych po przeszczepieniu nerki.

In the paper the authors tried to identify factors influencing prevalence and clinical course of cytomegalovirus (CMV) infection in kidney transplant patients. The study was performed in the group of 100 patients after cadaveric kidney transplant followed up in the Chair and Department of Nephrology, Collegium Medicum, Jagiellonian University in Krakow. CMV infection was demonstrated to occur more frequently and significantly earlier in the patients administered prednisone, cyclosporin A and mycophenolate mofetil, compared to the group treated with standard triple-drug-therapy (prednisone, cyclosporin A, azathioprine) or double-drug-therapy (prednisone, cyclosporin A). Higher serum levels of cyclosporin A did not increase prevalence of the infection but urged its onset. Risk for CMV infection was however higher in the group of patients treated for acute rejection episodes, especially with antilymphocyte preparations. No differences were shown in the immunological matching within HLA-A, -B and -DR antigens between the patients without features of CMV Infection and those treated for its active form. The infection occurred significantly more frequently in the recipients with HLA-A1 antigen than in those with HLA-A9 and -DR7. In patients with delayed transplanted kidney functioning, time of the infection onset and a number of its episodes were similar to the remaining population, however severity of the clinical course positively correlated with the duration of acute tubular necrosis (ATN). CMV infection occurred slightly more frequently in patients requiring transfusions compared to those not administered blood preparations. Among patients with AB blood type, active CMV infection occurred statistically less frequently, whereas in those with other blood types percentage of patients with/without CMV infection were comparable.

[Chronic inflammation of the colon: a diagnostic problem]. / Przewlekle odczyny zapalne jelita grubego--problem diagnostyczny.

Chronic inflammation of the colon and rectum with associated diarrhea has become more difficult to diagnose recently. Many etiological factors and different therapeutic outlines need to be carefully discussed and summarized. Based on most recent literature, authors present current data on etiology and specific (case-depended) diagnostic procedures in colitis and proctocolitis.

[Results of kidney transplantation in Krakow in 1992-2000]. / Wyniki przeszczepiania nerek w Krakowie w latach 1992-2000.

The aim of the study was an analysis of renal transplantation results in the Krakow Transplant Center during 1992-2000. The analysis concerned 94 cadaveric transplant recipients. The study group included 31 females aged 23 to 61 years (mean 40.4 years) and 63 males aged 16 to 60 years (mean 41.8 years). The time of pre-transplant renal replacement therapy ranged from 4 to 120 months (mean 32 months). The mean time of total ischaemia was 22 hours 20 minutes. The majority of the recipients had three identical antigens out of six typed. Most of the recipients were treated with three immunosuppressive drugs including: Cyclosporine A, Azathioprine and steroids. Immediately after kidney transplantation 25.6% of the patients had urine output and did not require dialysis. Acute renal failure (ARF) of the graft was observed in 73.2% recipients. The average number of hemodialysis sessions in patients presenting ARF was 10. Acute rejection was diagnosed in 41.5% of the patients. The most frequent complications were: CMV (cytomegalovirus) infection, UTI (urinary tract infection) and policytemia. In the study group 1-year survival rate of the patients was 97.8% and 1-year graft survival was 93.61%. The 5-year survival rates both in the patients and the grafts were very satisfactory (96.96% and 87.7% respectively).

[Acute neurologic rubella complications observed in children of southeastern Poland during rubella epidemics in 1986 and 1992]. / Ostre neurologiczne powiklania rózyczki obserwowane u dzieci z Polski poludniowo-wschodniej w czasie epidemii rózyczki w 1986 i 1992 r.

During the last two rubella epidemics in 1985-1986 and 1992, 24 children (15 boys and 9 girls) were hospitalized with acute neurological complications manifested in the first week of clinical symptoms of rubella. Average age of patients was 9 years (3-15 years). Acute rubella encephalitis (ARE) was diagnosed in 22 cases. Most of these patients had sudden loss of consciousness lasting from several hours to 12 days and convulsions during the first stage of the illness. Two patients developed retrobulbar neuritis which led to a significant impairment of sight in one of them. One child suffers epilepsy as a result of ARE. The remaining children did not develop lasting complications. During hospitalization, active infection by the rubella virus was confirmed in 20 children by detecting specific IgM antibodies in serum using the ELISA method. Comparing the ARE cases in 1986 and 1992 rubella epidemics revealed a change in clinical course. Earlier manifestations of neurological symptoms and more marked changes in CSF were observed. The issue of immunoprophylaxis is discussed; these measures only started in 1989 by vaccinating 13 and 14 year old girls. This method of prophylaxis will neither stop the transmission of the virus among children nor prevent the occurrence of periodic epidemics and rubella-related complications.

[The frequency of cytomegalovirus infection (CMV) in patients immunocompromised patients with malignant and nonmalignant blood disorders]. / Czestosc zakazenia wirusem cytomegalii (CMV) u chorych z niedoborem immunologicznym w przebiegu zlosliwych i niezlosliwych chorób krwi.

Studies on CMV infection were carried out in a group of 110 patients, aged from 15 to 78 (average 38), treated in the Department of Haematology. During the course of observation most of the patients were examined repeatedly. Diagnosis was based on serology (CFT, ELISA) and virus isolation from the clinical material. Primary infection (seroconversion) was confirmed in 4 (21%) out of 19 seronegative patients. Seropositive patients comprised 83 per cent. Active CMV infection was determined in 20 (22%) of those patients.

Rubella infection during pregnancy in the 1985-86 epidemic: follow-up after seven years.

The study covered 310 pregnant women from southern Poland who were exposed to rubella during the 1985-86 epidemic, none of whom had been vaccinated against rubella. Rubella specific antibodies were detected by hemagglutination-inhibition (HI) tests, and IgM antibodies by enzyme immunoassay (ELISA) (Organon Teknika). Clinical symptoms according to anamnesis were recorded. The consequences of serologically confirmed maternal rubella on the course of pregnancy and on fetal outcome were evaluated. IgM antibodies could be examined in only 10 newborns at delivery or in the first days of life. After seven years, follow-up studies of children born to infected mothers were done. The mental development of 14 of these children was evaluated with Terman-Merrill test. Among 310 women examined during pregnancy, rubella infection was confirmed serologically in 46 cases (14.8%). All but 3 of those had clinical symptoms. The course of pregnancy was observed in 36 of the infected mothers. Only 5 women (22.7%) who had the infection in the first trimester of pregnancy delivered a healthy child. The rate of complications in pregnancy among women infected in the second trimester was lower, and 8 (66.7%) bore healthy children. All the children born to mothers infected in the third trimester were healthy. Eight of the 10 newborns examined at delivery were IgM positive. Of 29 children congenital rubella syndrome (CRS) was confirmed in 5 cases, CRS compatible or CRS possible in 7 and 3 had congenital infection only confirmed serologically (IgM-positive) without defects or symptoms. Seventeen (58.6%) children were found healthy including the 3 who had congenital infection only. The mental development of 14 children at age 7 was assessed; 10 cases (72%) fell within rank II 130-85, and 4 (28%) were of borderline intelligence. The study indicates that congenital rubella is still a serious problem in Poland. Immunization was introduced only in 1988-89, for 13-year-old girls. Women of child-bearing age should be screened for rubella antibodies and those susceptible to rubella infection should be vaccinated.

Application of PCR in situ method for detection of human cytomegalovirus (HCMV) DNA.

Present HCMV diagnosis is relatively slow and inefficient. We applied PCR in situ to 15 samples of human salivary glands, 20 cytospins from blood, and 10 human fibroblast samples in order to detect the presence of HCMV DNA. The results indicate that PCR in situ is an effective and very sensitive method for detection of HCMV infections in variety of specimens.

A study on the antiviral action of a polyphenolic complex isolated from the medicinal plant Geranium sanguineum L. VIII. Inhibitory effect on the reproduction of herpes simplex virus type 1.

The antiviral activity of a polyphenolic complex, isolated from a Bulgarian medicinal plant Geranium sanguineum L., on the reproduction of herpes simplex virus type 1 was studied. The substance inhibited the intracellular stages of viral replication and exhibited a virucidal effect.

Cytomegalovirus infection in kidney transplant patients: clinical manifestations and diagnosis.

Cytomegalovirus (CMV) infection is an important cause of morbidity and mortality after solid organ transplantation. CMV infection after kidney transplantation was confirmed in 19 (54.3%) out of 35 patients. 16 of these (84.2%) developed CMV disease. CMV infection was diagnosed based on a fourfold or greater increase of anti-CMV IgG antibody titre, detection of CMV-IgM antibodies and/or virus isolation. Primary infection was observed in 3 patients, reactivation in 9 and an undefined type of infection in 7. In most patients (63%), infection was diagnosed in the first 2 months, and in 3 patients, after 3, 5 and 9 years following kidney transplantation. The most frequent symptoms of CMV disease were fever (58%), pneumonitis (26.3%) and enterocolitis (15.8%). In 53% of the patients, CMV infection co-occurred with other pathogens such as Candida albicans. Cryptococcus neoformans, bacteria or viruses (HBV, HCV, HSV). Treatment with polyvalent globulin (Sandoglobin) or hyperimmune globulin (Cytotect), in combination with ganciclovir in 7 patients, resulted in a regression of CMV disease.

Humoral beta-cell autoimmunity is rare in patients with the congenital rubella syndrome.

The congenital rubella syndrome (CRS) is associated with increased risk for diabetes and thyroid disease. However, the mechanisms by which the rubella virus may cause these diseases are poorly characterized. Previous studies were carried out before modern immunological methods were available. The present study aimed at evaluating whether autoimmune mechanisms are involved in the pathogenesis by analysing antibodies to biochemically characterized autoantigens. The incidence of clinical diabetes, thyroid disease, coeliac disease and related antibodies (islet cell antibodies, ICA; insulin autoantibodies, IAA; antibodies to the tyrosine phosphatase related IA-2 molecule, IA-2 A and glutamic acid decarboxylase, GADA; thyroid peroxidase, TPO; tissue transglutaminase, TTGA; and gliadin, AGA) and HLA risk genotypes were analysed in 37 subjects affected by or exposed to rubella during fetal life (mean age 22.5 years). One patient had diabetes and four patients had clinical hypothyroidism at the time of the examination. ICA, IAA, GADA or IA-2 A were not detected in any of the patients, while five patients tested positive for TPO antibodies. Coeliac disease or TTGA were not observed. Eight patients carried the HLA-DR3-associated HLA-DQB1*02-DQA1*05 haplotype. These results provide no evidence of an increased frequency of markers for humoral beta-cell autoimmunity in patients with CRS suggesting that diabetes in CRS may be caused by other than autoimmune mechanisms.