[Reed nevus and cutaneous melanoma in children: an immunohistochemical study]. / Nevus Rida i melanoma kozhi u detei. Immunogistokhimicheskoe issledovanie.

Certain difficulties arise in the differential diagnosis between Reed nevus that is common in children and adolescents and cutaneous melanoma that is extremely rare in patients of this age group. In addition to the classical histological examination, an immunohistochemical test for marker proteins is carried out to improve the accuracy of the diagnosis of pigmented skin neoplasms. OBJECTIVE: To evaluate the suitability of a set of proteins (cyclin D1, p16INK4a, and HLA class I antigens) in the differential diagnosis of Reed nevus and cutaneous melanoma in children and adolescents. MATERIAL AND METHODS: Three samples of pigmented skin neoplasms were taken during surgical treatment in patients at the Blokhin National Medical Research Center of Oncology: two samples of Reed nevus (from a 9-year-old girl and a 15-year-old boy) and a sample of cutaneous melanoma that had developed in an 8-year-old boy. The materials were presented by paraffin blocks. Immunohistochemical staining was performed using antibodies against cyclin D1, p16INK4a, and HLA class I antigens. RESULTS: Differences were found between nevus and melanoma in the expression of all three markers. Thus, the nuclear expression of cyclin D1, a proliferation activator, occurred in some cells in the nevus samples and in the vast majority of cells in the melanoma samples. The nuclear expression of the cell cycle inhibitor p16INK4a was dramatically reduced in melanoma compared to nevus. HLA class I antigens were detected on the surface of individual nevus cells, but were completely absent on the membranes and in the cytoplasm of melanoma cells, which could promote the evasion of this tumor from the body's immunological supervision. CONCLUSION: Immunohistochemical test for the proteins cyclin D1, p16INK4a, and HLA class 1 antigens is a promising approach to differentially diagnosing between Reed nevus and cutaneous melanoma in children and adolescents.

[Melanoma and Human Papillomaviruses: Is There an Outlook for Study?].

Melanoma is one of the most aggressive human malignant tumors. Its incidence and mortality are growing steadily. Ultraviolet irradiation is the main risk factor for melanoma involved in melanomagenesis. The probability of viral etiology of melanoma has been discussed. Human papillomaviruses (HPV) have been mentioned among candidates for its etiologic agents because some HPV types are the powerful carcinogens causing cervical cancer and other cancers. The review analyses the literature data on the association of melanoma with HPV Several groupsfound HPVin skin melanomas as well as in mucosa; viruses of high oncogenic risk were detected in some cases. For some organs the etiological role of high-risk HPV as inducers of invasive carcinomas is confirmed. These organs require special mention: cervix uteri, vulva, vagina, penis, anal region, and oral cavity. However in the majority of the studies in which viral DNA-positive melanomas were found, testing for viral genome expression was not done while this is the fact of primary importance. HPVare found in normal skin and mucous membranes thus creating justifiable threat of tumor specimen contamination with viral DNA in vivo. There are limited data on aggravation of the disease prognosis in papillomavirus-positive melanomas. However, any systematic observation of a sizeable patient group distinguished by that tumor type has not been performed yet. Viral E6 and E7 oncogenes of high-risk papillomaviruses were shown to be able to transform normal human melanocytes in vitro experiments. Thus, we can assume the presence of the association of melanoma with oncogenic HPV. The clinical significance of this problem is indisputable under the conditions of the steady increase in melanoma incidence and mortality rates in Russia and abroad. The problem requires further study.

Loss of Cell Differentiation in HPV-Associated Bladder Cancer.

Medical histories of 101 urothelial bladder cancer patients were compared with the results of morphological analysis and biomolecular detection of human papilloma viruses (HPV) in the tumor specimens. DNA of HPV16 (the major type of virus responsible for appearance of cervical carcinoma) was detected in 38 specimens, while mRNA of E6 and E7 oncogenes and E7 oncoprotein of HPV16 were observed in 13 specimens. HPV-positive bladder cancer was characterized by higher degree of cell anaplasia than HPV-negative cancer; in the primary bladder tumor, HPV was detected more often than in recurrent bladder cancer. These data attest to involvement of HPV16 in the genesis of bladder cancer. No correlations of HPV status of bladder tumor with patient's sex, age, and invasion into the muscle layer were revealed.

[Prostate cancer: papillomaviruses as a possible cause].

Prostate cancer (PC) incidence and mortality are steadily increasing. Causation of PC is not clearly understood; in particular, role of human papillomaviruses (HPV) is still disputable. The review contains analysis of literature data on possible participation of HPV powerful biological carcinogens, in PC genesis. PC incidence increase in persons with immunodeficiency indicates involvement of some infectious agent in the disease etiology. Several research groups communicated HPV DNA finding including that of oncogenic types in PC specimens (transrectal biopsies). There are limited data on the occurrence of oncogenic HPV 16 oncoprotein E7 in such specimens and on its unfavorable effect on disease prognosis. The successful attempt is known to transfect normal human prostate cells with oncogenic HPVDNA in vitro. Epidemiological data on associations of PC with HPV are controversial. It may result from the considered in the present review certain technical peculiarities of these studies. Controlfor serum antibodies to HPV E6 and E7 oncoproteins recognized to indicate HPV-positive tumor growth in an organism has not been performed yet in PC patients. DNA of oncogenic HPV is rather commonly found in organs adjacent to prostate--urethra, rectum, urinary bladder. In the study held in Russia on a group of healthy men examined for sexually transmitted diseases genitourinary HPVinfection was found in every second person; 42% of them harbored oncogenic HPV. Possible participation of oncogenic HPV in PC genesis deserves close attention and further study.

Complex of molecular genetic and immunohistochemical methods for detection of human papillomavirus in the bladder cancer epithelium.

A battery of tests for detection human papillomavirus DNA, mRNA corresponding to viral oncogenes, and viral oncoprotein E7 in cancer bladder urothelium was piloted in 35 samples of bladder cancer. DNA of human papillomavirus type 16 (causes cervical cancer) was found in 16 (46%) samples; E6/E7 oncogene transcript and E7 oncoprotein of human papillomavirus type 16 were detected in 10 and 7 human papillomavirus DNA-positive samples, respectively. These findings attest to association of bladder cancer with human papillomavirus in Russia.

Cellular expression of INK4a gene in cells of bladder cancer associated with human papilloma virus-16.

Hyperexpression of p16(INK4a) protein is an early marker of cervical cancer. Hyperexpression of INK4a gene encoding this protein at the level of mRNA and p16(INK4a) was detected in tumor cells of some patients with bladder cancer associated with human papilloma virus-16. However, in contrast to cervical cancer, this phenomenon in urothelial carcinomas does not correlate with expression of human papilloma virus-16 oncogenes E6 and E7.

[Are human papillomaviruses responsible for the occurrence of bladder cancer].

A female patient with recurrent bladder cancer underwent complex examination. The primary tumor removed in 2004 showed human papillomavirus (HPV) 16 DNA, mRNA corresponding to HPV16 oncogene E7, as well as HPV16 protein E7. The patient is a smoker who has been working at a chemical factory for over 20 years. During tumor recurrence in 2009, there was no DNA of high-risk HPV types in the cancer cells. HPV16 E7protein and cellular p 16(INK4alpha), an indicator of HPV-induced carcinogenesis, were not found. Colposcopy revealed no precancerous changes in the epithelium of the cervix uteri. The cervical epitheliocytes contained no high-risk HPV DNA, E7 and p16(INK4alpha) proteins. It seems expedient to continue in vitro studies of the possible role of HPV in urothelial carcinogenesis on an experimental model.

[Detection of oncoprotein E7 HPV16 in the cancer and normal urinary bladder urothelium].

Oncoprotein E7 HPV16 was detected by immunohistochemical staining with specific polyclonal antiserum [Fiedler et al., 2004] in 7 out of the 24 (29.2%) studied bladder cancer specimens. The result is in good agreement with the hypothesis that HPVs take part in the carcinogenesis of the urothelium. However, some of the observations made seem rather hard to be interpreted at present. The latter include the detection of E7 HPV16 in a small number of cancer cells in a few bladder cancer specimens being examined; the presence of this protein in the cytoplasm, rather in the cancer cell nuclei, and its detection in some morphologically normal bladder urothelial specimens from non-cancer patients. Thus, the hypothesis that HPVs are implicated in the carcinogenesis of the bladder urothelium deserves further verification.

[Polymethylene derivatives of nucleic bases with omega-functional groups: VII. Cytotoxicity in the series of N-(2-oxocyclohexyl)-omega-oxoalkyl substituted purines and pyrimidines].

New polymethylene derivatives of nucleic bases with a beta-diketo function in the omega-position were obtained by alkylation of uracil, thymine, cytosine, hypoxanthine, adenine, and N(2)-isobutyryl guanine with 2-omega-chloroal-kanoyl)cyclohexanones. The physical and chemical characteristics of the compounds synthesized and their effect on the K562 and HCT116 tumor cell lines were studied.

[Cell protein p16INK4a hyperexpression in epithelial malignancies induced by human papillomaviruses].

Protein p 16INK4a is an inhibitor of cyclin-dependent kinases Cdk4/6. It is encoded by the genome INK4a tumors unassociated with human papillomavirus (HPV) frequently inactivates; at the same type p16INK4a is commonly absent in the cells. In cancer of the cervix uteri (CCU) induced by HPV, INK4a hyperexpresses at the level of m-RNA and protein. Therefore p16INK4a is recommended for early diagnosis of CCU. Attention to p16INK4a additionally increases as there is increasing evidence for the implication of HPV in the etiology of some other cancers. The review considers the mechanisms of hyperexpression of p16INK4a in HPV-induced cancers. There is new evidence for the diagnostic value of a positive test for p16INK4a. Moot points of interpreting the results of immunohisto-/cytochemical detection of p16INK4a are discussed.

[Expression of p16INK4a in various cancer cells]. / Ekspressiia belka P16ink4a v kletkakh nekotorykh rasprostranennykh form raka.

The p16INK4a protein was detected by means of monoclonal antibodies to this protein in the cells of some carcinomas: that of the lungs (17 samples), urinary bladder (6 samples) and mammary gland (4 samples) as well as in the cells of three cell lines from of human uterine cervix carcinoma: SiHa (containing high risk HPV genome), C33A and HT3 (both HPV-negative but have RB mutations in RB gene). Lung carcinoma samples were very heterogenous by the part of cells expressing p16INK4a. High content of this protein was found in all 6 samples of transient cell urinary bladder carcinoma and in 1 sample of mammary gland ductal carcinoma. Cells of all three cell lines also contained p16INK4a. Thus, hyperexpression of this protein is not specific for only HPV-positive cancer of the uterine cervix. The protein presence in cancer cells seems to be an indicator of gene RB mutation or other disturbances of RB pathway.

[Interaction of viral and cellular genes in cervical tumors]. / Vzaimodeistvie virusnykh i kletochnykh genov v opukholiakh sheiki matki.

The results obtained in the Laboratory of Molecular Biology of Viruses, CRC carried out in the framework of the Human Genome program and devoted to the study of the activity of cell and viral genes in cervical cancer are summarized. DNA of human papillomaviruses persists in tumors both in episomal and integrative forms. Integration may occur in different regions of chromosomes. Viral transforming genes E6 and E7 are always present in tumor cells, while antibodies to these proteins are detected only in approximately 30% of patients. Loss of heterozygosity is detected on long and short arms of chromosome 6; some such cases are manifest already at the early stages of tumor progression, while others are typical of the late stages. Several genes that are potentially involved in tumorigenesis and are subject to hypermethylation in CpG islands were identified. Methylation of several genes is observed in approximately 30% of tumors. Tumor progression is associated with increased expression of p16ink4a, an inhibitor of cyclin-dependent kinases.

[Expression of the protein marker p16INK4a in the cervix uteri cancer]. / Ekspressiia belkovogo markera p16INK4a v rake sheiki matki.

Immunohistochemical study was carried out of 18 cervical carcinomas (13 squamous and 5 adenomatous) and of 3 cases of cervical intraepithelial dysplasia. Formalin-fixed paraffin-embedded tissue samples from biopsies as well as from surgical material were used. Staining was performed with monoclonal antibodies to protein p16INK4a. Cytologic smears of epithelial cervical cells from 7 healthy women were taken as a negative control. The reference group consisted of 5 cancer patients with other tumors (breast cancer, B-cell lymphoma). Overexpression of p16INK4a was registered in cervical cancer.

[Search for biological remedies for fungi of the Candida genus]. / Poisk biologicheskikh sredstv protiv gribov roda Candida.

51 bacterial strains were studied with a view to find out anticandidial activity; of these, 15 strains were found to have activity, more on less expressed with respect to fungi of the genus Candida. In most cases the anticandidial activity of an antagonist strain was manifested to a similar degree against several Candida strains. The preliminary analysis of antifungal substances produced by microbial strains makes it possible to suggest their antibiotic nature.

[General problems in the study of the mutagenic properties of drugs]. / Obshchie voprosy izucheniia mutagennykh svoistv lekarstvennykh sredstv.

The article deals with the results characterizing the state of Russian and foreign elaborations in the study of genotoxic effects of drugs and immunobiological agents. Particular attention is placed on general problems of strategy and tactics in the study of mutagenicity of drugs the solution of which is insufficiently substantiated or insufficiently discussed in the literature. The perspective problems of research into the field of genotoxicity of drugs are discussed.

The influence of two carotenoid food dyes on clastogenic activities of cyclophosphamide and dioxidine in mice.

The influence of the food dyes E160e (beta-apo-8'-carotenal in an oil suspension) and E160a (beta-carotene in an oil suspension) on clastogenic effects of cyclophosphamide (CP) and dioxidine (DN) was investigated. Chromosome damage in the bone marrow of C57BL/6 mice was reported. The following protocols were used: (1) simultaneous single administration of the dye and the mutagen and the subsequent animal sacrifice within 24 hr; (2) a 4-day pretreatment with the dye (daily administrations) followed with simultaneous injection of the dye and the mutagen on the 5th day 24 hr before sacrifice; (3) daily co-administration of the dye and the mutagen for 5 days with sacrifice 6 hr after the last administration. CP at a dose of 30 mg/kg and DN at 300 mg/kg were injected intraperitoneally; the dyes at doses of 0.5, 5 and 50 mg/kg were given orally. Under all the protocols applied, E160e at a dose of 50 mg/kg caused a significant reduction of both DN and CP effects. At 5 mg/kg this dye reduced the effects of the mutagens only under the pretreatment regimen. Pretreatment with E160a at doses of 5 and 50 mg/kg resulted in a meaningful reduction of the DN effect. Under the combined treatment with mutagens this dye reduced both CP and DN effects.

[Genotoxicity of vaccines: an unresolved problem of ecological genetics]. / Genotoksichnost' vaktsin: ékologo-geneticheskaia problema s nezavershennym resheniem.

The problem of potential remote consequences of vaccinations still remains to be solved. Data on the effects of immunobiological preparations, including vaccines, employed for preventive mass immunization on certain hereditary structures are reviewed. Many of these preparations are genotoxic. Prospects for diminishing the genetic risks of vaccinations are discussed.

[Control of genetic consequences of vaccinations: electron microscopic analysis of murine synaptonemal complexes]. / Kontrol' geneticheskikh posledstvii vaktsinatsii: élektronno-mikroskopicheskii analiz sinaptonemnykh kompleskov myshi.

The ability of two inactivated bacterial vaccines, from Proteus vulgaris and Klebsiella pneumoniae, to injure synaptonemal complexes (SCs) was studied by means of electron microscopy. The preparations were given intraperitoneally to C57BL/6J male mice during five successive days and testes were fixed 24 h after the last injection. Cyclophosphamide was used for positive control. The vaccine from Klebsiella given at subtoxic dose, which was about 200 times higher that that given to people during vaccinations, induced the 10-fold rise in the frequency of SC abnormalities in murine 1-storger spermatocytes. Breaks of SCs and of single lateral elements of SCs predominated over other types of vaccine-induced anomalies. According to the preliminary data, vaccine from Proteus at subtoxic dose showed no damaging SC activity. The results of the given study are discussed in connection with the negative data obtained earlier when genotoxicity of these two vaccines had been studied in the Ames test, in routine investigations of bone marrow metaphases of vaccinated mice as well as under light microscopy of their SCs.

[The mutagenicity of a new multicomponent vaccine made from the antigens of Klebsiella pneumoniae, Staphylococcus aureus, Proteus vulgaris and Escherichia coli]. / Issledovanie mutagennosti novoi polikomponentnoi vaktsiny iz antigenov Klebsiella pneumoniae, Staphylococcus aureus, Proteus vulgaris i Escherichia coli.

Inactivated bacterial vaccine, containing K. pneumoniae, S. aureus, P. vulgaris and E. coli antigenic complexes were tested for mutagenicity in the test described by Ames et al. and in vivo, in experiments on mice. In Salmonella typhimurium cells, strain TA-98 and TA-100, the preparation (5-75 mg/ml) did not increase the frequency of reversions and histidine-independence either in direct experiments or after metabolic activation with rat liver homogenate. In experiments on mice the vaccine (3.3 mg/kg and 33 mg/kg) did not induce chromosomal anomalies in spermatogonia. In all experiments the mutagens used for positive control produced a mutagenic effect.

[The chromosome-damaging action of 2 chemical vaccine contaminants on mice]. / Issledovanie povrezhdaiushchego kromosomy deistviia dvukh khimicheskikh kontaminantov vaktsin na myshakh.

The cytogenetic effects of two chemical agents, hydroxylamine used for the destruction of bacterial cells and thimerosal added to many immunobiological preparations as preservative, were studied in vivo by their action on the marrow cells of C57BL/6J mice. The preparations under study, when injected intraperitoneally in a wide range of doses, including subtoxic ones, induced no chromosomal aberrations. At the same time cyclophosphamide, an antitumor cytostatic agent used for positive control, produced a pronounced damaging effect on chromosomes.