[Cytological assessment of the nasal mucosa in children with adenoidal hypertrophy. Part I]. / Ocena cytologiczna blony sluzowej nosa u dzieci z przerostem migdalka gardlowego. Czesc I.

Investigation was performed on 68 children (24 girls and 44 boys) aged of 3-14, who was qualified by paediatricians for cytological evaluation of the nasal mucosa. They had undergone treatment for chronic upper respiratory tract infection and had clinical symptoms of the adenoidal hypertrophy. The adenoidal hypertrophy in children was confirmed by x-ray of nasopharynx. Seven children had previously undergone adenoidectomy and 3 had recurrent otitis. In the investigated group of children the majority of cytograms contained high level of neutrophils (> 50% of the total number of cells). Only 2 children showed normal cytogram. Discovery of substantial eosinophilia of the nasal mucosa (11.76% of investigated children) indicates that allergy diagnosis should be performed in children with adenoidal hypertrophy.

[Cytological assessment of nasal mucosa in children with adenoidal hypertrophy. Part II]. / Ocena cytologiczna blony sluzowej nosa u dzieci z przerostem migdalka gardlowego. Czesc II.

In this investigation we would like to evaluate what changes in cytology of the nasal mucosa occur after surgical removal of adenoids. The investigation was performed on 25 children (10 girls and 15 boys) aged 3-13. Material for the cytological evaluation was obtained from the nasal concha. The children were tested twice: before and one month after the surgery. Cytograms of the nasal mucosa of children with adenoidal hypertrophy before the surgery showed increased number of neutrophils. Cytological normalisation of the nasal mucosa occurs in one month after the surgical removal. In two children percentage of eosinophils in cytograms increased and percentage of neutrophils remained significant after the surgery. Cytological assessment of the nasal mucosa performed before and after surgical removal of the hypertrophied adenoid is helpful in evaluation of the accomplished therapy and may indicate what are the reasons of its failure.

Oncogenic potential of human neurotropic virus: laboratory and clinical observations.

Cancer is a multi-step disease involving a series of genetic alterations that result in the loss of control of cell proliferation and differentiation. Such genetic alterations could emerge from the activation of oncogenes and the loss or malfunctioning of tumor suppressor gene activity. Our understanding of cancer has greatly increased through the use of DNA tumor viruses and their transforming proteins as a biological tool to decipher a cascade of events that lead to deregulation of cell proliferation and subsequent tumor formation. For the past ten years our laboratory has focused on the molecular biology of the human neurotropic papovavirus, JCV. This virus causes progressive multifocal leukoencephalopathy, a fatal neurodegenerative disease of the central nervous system in immunocompromised patients. JCV is a common human virus that infects more than 80% of humans but does not induce any obvious clinical symptoms. The increased incidence of acquired immune deficiency syndrome and the use of immunosuppressive chemotherapy have dramatically raised the incidence of PML. The coincidental occurrence of malignant astrocytes and oligodendrocytes in PML patients, coupled with the induction of glioblastoma in JCV-infected nonhuman primates, provides intriguing speculation on the association between JCV and CNS malignancies. In this report we discuss clinical data and laboratory observations pointing to the direct involvement of JCV in cancer.

Association of SV40 with human tumours.

SV40 was discovered as a contaminant of poliovirus vaccines that were inadvertently administered to millions of people in Europe and the United States between 1955 and 1963. Shortly afterwards, SV40 was proven to be oncogenic in rodents and capable of transforming human and animal cells in vitro. The possibility that SV40 might cause tumours in humans thus became a subject of scientific and public interest and scrutiny. However, largely due to a lack of significant epidemiological evidence, interest in assessing SV40's potential carcinogenic role in humans diminished. Recently, many laboratories have reported the presence of SV40-like DNA in a high proportion of human mesotheliomas, ependymomas and osteosarcoma (the three main types of tumours caused by virus in hamsters), renewing the question whether SV40 might be a human tumour virus. Molecular data from these studies are reviewed to re-evaluate the potential role of SV40 as a human carcinogen.

Reactivation of human neurotropic JC virus expressing oncogenic protein in a recurrent glioblastoma multiforme.

Examination of the primary tumor of glioblastoma multiforme and its recurrence for their association with JC virus revealed that, while the viral genome is present in both initial and recurrent tumors, expression of the viral oncoprotein T-antigen occurs only in the recurrent tumor cells. Accordingly, the level of inducible cellular transcription factors, including the p65 subunit of NF-kappaB and YB-1, which have the ability to stimulate JCV gene expression, was found to be higher in the recurrent tumor cells. These observations suggest that induction of the regulatory factors after resection of the primary tumor may have reactivated JC virus gene expression and led to redevelopment of the tumor in brain.

[Cytologic evaluation of nasal mucosa in healthy newborns]. / Ocena cytologiczna blony sluzowej nosa u zdrowych noworodków.

Cytological evaluation of the nasal mucosa is one of the methods used in the diagnosis of upper respiratory diseases. Cytological examination of the nasal mucosa in very young children have sporadically been performed. In our study the evaluation was performed in 4 hospitals in 3 towns (Warsaw, Czestochowa, Bielsko-Biala) in Poland. Cytograms of 105 infants age 1-7 days of both sexes were evaluated. All the infants were born naturally, on time and were healthy, according to the opinion of physicians. The course of pregnancies and birth-giving were considered as normal. There were no history of allergic diseases in the families in each case. Cytograms of the nasal mucosa of 1-7 days old infants differ from these of adults and older children. Either columnar cells or neutrrophils are prevalent in cytograms of the healthy newborn children. Goblet cells do not occur. Cytological evaluation of nasal mucosa as non-invasive and easy to perform examination can be very useful in screening of the very young children from the group with a risk of respiratory system disorders including allergic diseases.

Identification of a novel p53 mutation in JCV-induced mouse medulloblastoma.

Medulloblastoma, a malignant invasive tumor of the cerebellum, is one of the most common neoplasms of the nervous system in children. Utilization of the human neurotropic virus JC virus (JCV) early gene T-antigen allowed the development of a transgenic animal that models human medulloblastoma. Here we describe the characterization of two distinct populations of cells derived from the JCV-induced mouse medulloblastoma. Results from immunohistochemical and biochemical studies revealed the expression of T-antigen in some but not all tumor cells. In T-antigen-producing cells, T-antigen was found in association with wild-type p53 and pRb, two tumor suppressors that control cell growth and differentiation. In cells that lack expression of T-antigen, a novel mutant p53 with a deletion between residues 35 and 123 was detected. Morphological differences were observed between the two populations of cells, though there was no significant difference in their growth rates. However, subcutaneous transplantation of the T-antigen-positive, but not T-antigen-negative, cells resulted in the development of massive tumors in experimental animals. In light of earlier reports on the association of JCV with human medulloblastoma, the mouse cell lines described in this study may provide a valuable tool for deciphering the pathways involved in the formation and progression of medulloblastoma.

Detection of human neurotropic JC virus DNA sequence and expression of the viral oncogenic protein in pediatric medulloblastomas.

Medulloblastoma represents greater than 25% of childhood intracranial neoplasms and is considered a highly malignant tumor. This tumor, which arises predominantly in the cerebellar vermis, preferentially affects children between the ages of 5 and 15. Although the etiology of medulloblastomas in humans remains unknown, results from several experiments have indicated that the human neurotropic JC virus (JCV) is able to induce cerebellar neoplasms in rodents that exhibit a phenotype similar to that of human medulloblastomas. JCV is a polyomavirus that is widespread in the human population, with infection occurring most frequently in early childhood. In this study, we have examined the possible association of JCV with human medulloblastomas. By using PCR techniques we demonstrate that 11 of 23 samples of tumor tissue contain DNA sequences corresponding to three different regions of the JCV genome. More importantly, we demonstrate the presence of DNA sequences encoding the N- and C-terminal regions of the JCV oncogenic protein, T antigen, in 11 of 23 samples and the production of T antigen in the nuclei of 4 samples of tumor tissue. These observations provide evidence for a possible association of JCV with human medulloblastomas.

Human neurotropic JC virus early protein deregulates glial cell cycle pathway and impairs cell differentiation.

Progressive multifocal leukoencephalopathy (PML), a human demyelinating disease of the central nervous system (CNS), is induced upon replication of the human neurotropic virus, JCV, in glial cells. Similar to other polyomaviruses, replication of JCV is initiated and orchestrated by the viral early protein, T-antigen, and results in the cytolytic destruction of oligodendrocytes, the subset of glial cells responsible for myelin production, and the appearance of bizarre astrocytic glial cells in affected individuals. Earlier results from studies in transgenic animals have suggested that in the absence of viral replication, expression of JCV T-antigen induces pathology consistent with hypomyelination of the brain. These observations suggest that JCV T-antigen has the ability to deregulate oligodendrocyte and perhaps astrocyte function in the CNS. Here we demonstrate that expression of JCV T-antigen in the bipotential glial cell line, CG-4, severely affects the ability of these cells to differentiate toward oligodendrocyte and astrocyte lineages as evidenced by their distinct morphological changes. Examination of the activity of cell cycle regulatory proteins including cyclins and their associated kinases reveals that in the absence of T-antigen, differentiation of CG-4 cells toward astrocytes and oligodendrocytes is accompanied by a decline in cyclin E, cdk2, cyclin A, and cyclin B activity. In contrast, cdc2 activity increased upon CG-4 differentiation. In T-antigen-producing cells, distinct variations in the activity of several cyclins was observed. For example, while the activity of cdk2 and cyclin E was enhanced in T-antigen expressing astrocytes compared to their levels in control cells, the activity of cdc2 was decreased in this cell type. In oligodendrocytes, expression of T-antigen decreased the activity of several cyclins and cdks including cyclin E and cdc2. On the other hand, the level of expression and activity of cyclin A was increased. Thus, it is evident that JCV T-antigen deregulates several important cell cycle regulators during CG-4 differentiation, and these alterations may contribute to the process of cell growth and differentiation in glial cells. The importance of our findings with regard to the neuropathogenesis of PML is discussed.

Human ubiquitous JCV(CY) T-antigen gene induces brain tumors in experimental animals.

JCV is a papovavirus which is widespread in the human population. The prototype Mad-1 variant of JCV induces a fatal demyelinating disease of the central nervous system (CNS) called Progressive Multifocal Leukoencephalopathy (PML) in immunosuppressed individuals. The unique tropism of JCV (Mad-1) to the CNS is attributed to the tissue-specific regulation of the viral early promoter which is responsible for the production of the viral regulatory protein, T-antigen. The archetype form of this virus, JCV(CY), which has been repeatedly isolated from the urine of PML and non-PML individuals, is distinct from JCV(Mad-1) in the structural organization of the regulatory sequence. To characterize the tissue specific expression of JCV(CY) and to investigate its potential in inducing disease, transgenic mice containing the early region of JCV(CY) were generated. Some of these mice between 9-13 months of age exhibited signs of illness as manifested by paralysis of rear limbs, hunched posture, and poor grooming. Neuropathological examination indicated no sign of hypomyelination of the brain, but surprisingly, revealed the presence of primitive tumors originating from the cerebellum and the surrounding brain stem. The tumor masses also infiltrated the surrounding tissue. Results from RNA and protein studies revealed a high level of T-antigen mRNA expression in hindbrains of clinically normal and affected transgenic mice. However, higher levels of T-antigen RNA and protein were detected in brains of the animals exhibiting severe illness. The close resemblance of JCV(CY) induced tumor in transgenic mice to the human medulloblastoma/primitive neuroectodermal tumor (PNETs) in location, histologic appearance, and expression of marker proteins strongly suggests the utility of this novel animal model for the study of human brain tumors.

Oncogenic potential of human neurotropic papovavirus, JCV, in CNS.

The human polyomavirus, JCV, is the causative agent of Progressive Multifocal Leukoencephalopathy (PML), a fatal human demyelinating disease. PML results from the cytolytic destruction of oligodendrocytes, the myelin-producing cells of the nervous system. JCV has also been shown to be tumorigenic in several animal models. Transgenic mice expressing the JCV early protein, T-antigen, develop poorly differentiated neural crest origin tumours. Intracerebral inoculation of JCV into newborn hamsters induces medulloblastomas, astrocytomas, and primitive neuroectodermal tumours. Further, inoculation of the virus into the brains of non-human primates, owl and squirrel monkeys, results in astrocytomas and glioblastoma multiforme. Several case reports have associated JCV with human CNS tumours in patients with concomitant PML, and one such report has detected JCV in a glial tumour in the absence of PML. The induction of neural origin tumours by JCV has been studied in transgenic mice harbouring the early genome of the virus. Alterations in the level and function of tumour suppressor proteins p53 and Rb, as well as associated cell cycle regulators, have been detected in tumour tissue from JCV T-antigen transgenic mice. Possible mechanisms by which JCV may exert its oncogenic potential by alteration of cellular growth control pathways in both humans and experimental animals are discussed.

A multi-institutional study confirms the presence and expression of simian virus 40 in human malignant mesotheliomas.

Exposure to the carcinogen asbestos is a major factor in the development of malignant mesothelioma. However, not all mesotheliomas are associated with asbestos exposure, and only a small minority of people exposed to asbestos develop mesothelioma. Therefore, the identification of the cofactors that render certain individuals more susceptible to asbestos or that cause mesothelioma in people not exposed to asbestos has been a major priority of the International Mesothelioma Interest Group. The possible association of SV40 with mesothelioma was recently discussed in a special session at the Fourth International Mesothelioma Interest Group Conference, and it was decided to conduct a multi-institutional study to independently verify the presence of this tumor virus in mesotheliomas. We report the results of this investigation: (a) DNA and protein analyses revealed SV40 sequences and SV40 large T antigen expression in 10 of 12 mesotheliomas tested (83%); and (b) electron microscopy demonstrated variable amounts of asbestos fibers in 5 (71%) of 7 corresponding lung tissues available for analysis. Our results demonstrate that SV40 DNA is frequently present and expressed in mesotheliomas in the United States. Because our data demonstrate that some patients test positive for both SV40 and asbestos, the possibility that these two carcinogens interact should be investigated in future studies.

[Disturbance of the normal cell cycle by T antigens of SV40 viruses and JC cause some tumors]. / Zaburzenia prawidlowego przebiegu cyklu komórkowego przez antygeny T wirusów SV40 i JC przyczyna niektórych nowotworów.

The mechanism that may lead to tumor formation in SV40 or JCV infected tissues based on previously reported interactions between T antigens and two factors (p53 and pRb) controlling cell cycle has been discussed. p53 is a known tumor suppressor protein that delays S phase entry causing cell arrest in G1 phase or apoptosis when the DNA damage occurs. Phosphorylation of pRB releases E2F family proteins that activate genes encoding S phase promoting factors. Binding of T antigens to pRB induces tumor formation, whereas tumor proliferation requires knockout of p53 activity.

MyEF-3, a developmentally controlled brain-derived nuclear protein which specifically interacts with myelin basic protein proximal regulatory sequences.

Myelin basic protein (MBP) gene contains the upstream regulatory sequence that confers cell type- and stage-specific transcription to MBP expression in oligodendrocytes during brain development. The MB1 regulatory motif located between -14 to -50 with respect to transcription start site binds to a brain derived nuclear protein and plays an important role in transcriptional activation of the MBP promoter in transfection assay. Here, we report the isolation of a recombinant cDNA clone, termed myelin expression factor-3, (MyEF-3) from a mouse brain expression library that encodes a novel protein which interacts with the MBP MB1 domain. Computer assisted evaluations of the MyEF-3 sequence revealed several interesting features including four sites for phosphorylation by casein kinase II, a transmembrane domain at the N-terminus, a nuclear localization signal and a Zinc finger domain at the carboxyl terminal. Results from Western and band shift assays indicate that MyEF-3 binds efficiently to double-stranded MB1 as well as the single-stranded non-coding strand of MB1. The use of short DNA fragments encompassing the nucleotide base substitutions across the MB1 domain in competition band shift assay revealed that the ten nucleotide sequence, 5'-GCCTGTCTTT-3' is important for binding of MyEF-3 to DNA. Results from Northern blot studies demonstrate that expression of MyEF-3 is restricted to brain and developmentally regulated during brain maturation. The biological importance of MyEF-3 in the cell type- and stage-specific expression of MBP during brain development is discussed.

Role of cell cycle regulators in tumor formation in transgenic mice expressing the human neurotropic virus, JCV, early protein.

Transgenic mice harboring the early genome from the human neurotropic JC virus, JCV, develop massive abdominal tumors of neural crest origin during 6-8 months after birth and succumb to death a few weeks later. The viral early protein, T-antigen, which possesses the ability to transform cells of neural origin, is highly expressed in the tumor cells. Immunoblot analysis of protein extract from tumor tissue shows high level expression of the tumor suppressor protein, p53, in complex with T-antigen. Expression of p21, a downstream target for p53, which controls cell cycle progression by regulating the activity of cyclins and their associated kinases during the G1 phase, is extremely low in the tumor cells. Whereas the level of expression and activity of cyclin D1 and its associated kinase, cdk6, was modest in tumor cells, both cyclin A and E, and their kinase partners, cdk2 and cdk4, were highly expressed and exhibited significant kinase activity. The retinoblastoma gene product, pRb, which upon phosphorylation by cyclins:cdk induces rapid cell proliferation, was found in the phosphorylated state in tumor cell extracts, and was detected in association with JCV T-antigen. The transcription factor, E2F-1, which dissociates from the pRb-E2F-1 complex and stimulates S phase-specific genes upon phosphorylation of pRb and/or complexation of pRb with the viral transforming protein, was highly expressed in tumor cells. Accordingly, high level expression of the E2F-1-responsive gene, proliferating cell nuclear antigen (PCNA), was detected in the tumor cells. These observations suggest a potential regulating pathway that, upon expression of JCV T-antigen, induces formation and progression of tumors of neural origin in a whole animal system.

[Cell response in the nasal mucosa in patients with hay fever during plant pollen season]. / Odpowiedzi komórkowe blony slzowej nosa u chorych z pylkowica w okresie pylenia roslin.

The aim of the present study was to determine whether different reactions of the nasal mucosa can be a result of a natural provocation with allergen. During the pollen season of anemorphilous plants the nasal mucosa may trigger the following reactions allergic inflammation or non-specific secretion.

[Steroid therapy and surgical treatment of nasal polyps: cytological analysis]. / Steroidoterapia i leczenie operacyjne polipów nosa w ocenie cytologicznej.

We have studies influence of steroids applied topically (intranasally) in patients with polyps (both operated on and not operated) on the cytology of nasal mucosa in conjunction with the level of serum ECP.

[Glucocorticosteroids: mechanism of action, pharmacological effects, pharmacokinetics and adverse effects]. / Glikokortykosteroidy--mechanizm dzialania, efekty farmakologiczne, farmakokinetyka i dzialania niepozadane.

The present review summarized the state of our knowledge on glucocorticosteroids, their mechanisms of action, pharmacological effects, pharmacokinetics and adverse effects. Oral steroids are very effective in treatment of asthma, however, when given in more than low doses and for a prolonged time, they may have quite serious side effects. The pharmacology of inhaled steroids is described. New inhaled glucocorticosteroids like budesonide and flunisolide are readily absorbed from the airway mucosa into the blood but are rapidly biotransformed in the liver into au inactive metabolite. Obviously the greatly reduces the risk of systemic side effects that exist with older glucocorticosteroids, e.g. dexamethasone.

[Eight-year long study of the usage of flunisolide (syntaris) at the Otolaryngological clinic at the Warsaw Academy]. / Osmioletnie doswiadczenia Kliniki Otolaryngologii Akademii w Warszawie w stosowaniu flunisolidu (Syntaris).

Brief information about clinical experience on flunisolide in nasal pathology treatment is presented in this paper. The authors own observations are also included. They describe some cases histories as to how flunisolide can act positively and negatively at on nasal and paranasal clinical pathology.