KCNH1 potassium channels are expressed in cervical cytologies from pregnant patients and are regulated by progesterone.

Potassium voltage-gated channel, subfamily H (eag-related), member 1 (KCNH1) potassium channels are potential tumour markers and cancer therapeutic targets and are up-regulated by oestrogens and human papilloma virus (HPV) oncogenes. However, the role of KCNH1 in normal tissues is poorly understood, and its expression in pregnancy is unknown. We wondered whether KCNH1 channels are expressed in cervical cells from pregnant patients and whether progesterone (P4) regulates KCNH1. The association with HPV was also investigated. KCNH1 protein expression was studied by immunocytochemistry in liquid-based cervical cytologies; 93 samples were obtained from pregnant patients at different trimesters, and 15 samples were obtained from non-pregnant women (controls). The presence of HPV was studied by PCR with direct sequencing and nested multiplex PCR. HeLa cervical cancer cells were transfected with human progesterone receptor-B (PR-B) and treated with P4. KCNH1 mRNA expression in these cultures was studied by real-time PCR. KCNH1 protein was detected in 100% of the pregnancy samples and in 26% of the controls. We found 18 pregnant patients infected with HPV and detected 14 types of HPV. There was no association between the percentage of cells expressing KCNH1 and either the presence or type of HPV. P4 induced KCNH1 mRNA and protein expression in cells transfected with human PR-B. No regulation of KCNH1 by P4 was observed in non-transfected cells. We show for the first time the expression of an ion channel during human pregnancy at different trimesters and KCNH1 regulation by P4 in human cells. These data raise a new research field for KCNH1 channels in human tissues.

High prevalence of oncogenic human papillomavirus in the genital tract of women with human immunodeficiency virus.

OBJECTIVE: The goal of this study was to determine the prevalence of human papillomavirus (HPV) and squamous intraepithelial lesions (SILs) in women infected with human immunodeficiency virus (HIV) in Mexico. METHODS: Cases included women who were positive for human immunodeficiency virus (HIV) and accepted to participate. There were two control groups in this study: group A, heterosexual partners of HIV+ men; group B, commercial sex workers. Gynecologic examination was performed in all participants. Also, a cervical smear with colposcopy and a sample for detection of HPV DNA by polymerase chain reaction (PCR) were obtained in all subjects, as were CD4+ counts. Relative risks (RR) and 95% confidence interval were calculated. RESULTS: Eighty-five HIV+ women agreed to participate in this study; the route of HIV infection was heterosexual in 78.8%; transfusion in 8.2%; paid donors in 3.5%; and 9.4% unknown. A total of 9 controls were included: 4 from group A and 5 from group B. HPV DNA was detected by PCR in 57 (69%) cases and in 26 (29%) controls from both groups (P < 0.0001). The RR of HPV infection was 5.5 (2.7-11.5). Also, a significant difference in the prevalence of high-risk HPV types was observed between cases and controls, RR = 12.8 (4.07-42.9). These associations were independent of CD4+ counts and antiretroviral therapy. No association was observed between HIV infection and the risk for high-grade SIL. CONCLUSIONS: We observed a high prevalence of oncogenic HPV types in HIV-positive women. These women should be screened regularly for early diagnosis of premalignant lesions and prevention of cervical cancer.

[Molecular variants of human papillomavirus (HPV) types 16 and 18 in adenocarcinomas of the cervix]. / Variantes moleculares de virus papiloma humano (HPV) tipos 16 y 18 en adenocarcinomas de cérvix.

Human Papillomavirus (HPV), placebo clinical trial particularly types 16 and 18, are considered human carcinogens since an etiological association has been demonstrated between these viruses and the development of cervical cancer. While the viral role in squamous carcinoma has been largely studied, the information available on adenocarcinoma is scarce, partly because of its lower frequency. In this paper we investigated the presence of HPV types and intratype variants in adenocarcinomas of the cervix. A total of 23 archive samples, fixed and paraffin embedded biopsies, were included. The detection and viral typing was performed by generic PCR and subsequent single stranded conformational polymorphism analysis (SSCP). Genetic variability was investigated in a 450 bp-fragment corresponding to L1 gene by post-PCR direct sequencing. We detected 11 HPV 16 positive samples (9 prototypes and 2 variants: 1 European and 1 Asiatic-American), 10 HPV 18 (9 prototypes and 1 European variant), 1 HPV 31 and 1 negative. The high risk HPV association with this neoplasia was confirmed with a high prevalence (43%) of HPV 18, (but) without predominance over the other types as previously published. The demonstrated variability in L1 protein epitopes originated aminoacidic changes which could have implications on the immune response and therefore should be considered in a vaccine design.

Variantes moleculares de virus papiloma humano (HPV) tipos 16 y 18 en adenocarcinomas de cérvix. / [Molecular variants of human papillomavirus (HPV) types 16 and 18 in adenocarcinomas of the cervix]

Human Papillomavirus (HPV), placebo clinical trial particularly types 16 and 18, are considered human carcinogens since an etiological association has been demonstrated between these viruses and the development of cervical cancer. While the viral role in squamous carcinoma has been largely studied, the information available on adenocarcinoma is scarce, partly because of its lower frequency. In this paper we investigated the presence of HPV types and intratype variants in adenocarcinomas of the cervix. A total of 23 archive samples, fixed and paraffin embedded biopsies, were included. The detection and viral typing was performed by generic PCR and subsequent single stranded conformational polymorphism analysis (SSCP). Genetic variability was investigated in a 450 bp-fragment corresponding to L1 gene by post-PCR direct sequencing. We detected 11 HPV 16 positive samples (9 prototypes and 2 variants: 1 European and 1 Asiatic-American), 10 HPV 18 (9 prototypes and 1 European variant), 1 HPV 31 and 1 negative. The high risk HPV association with this neoplasia was confirmed with a high prevalence (43

) of HPV 18, (but) without predominance over the other types as previously published. The demonstrated variability in L1 protein epitopes originated aminoacidic changes which could have implications on the immune response and therefore should be considered in a vaccine design.

Neoadjuvant chemotherapy interferon for epidermoid penile cancer: a case report

Antecedentes. El carcinoma avanzado de pene es un tumor con un pronóstico malo con el tratamiento estándar por lo que la quimioterapia neoadyuvante se ha estado evaluando para mejorar la preservación del órgano y la supervivencia. Caso clínico. Paciente de 56 años visto en el Instituto Nacional de Cancerología en noviembre de 1993 siendo diagnosticado de cáncer de pene T3 N3 MO. El tratamiento consistió en cisplatino 100 mg/m2 cada 21 días por cinco ciclos alternados con metotrexato intravenoso a dosis de 250 mg/m2 más rescate con leucovorín oral a dosis de 10 mg/m2 cada ocho horas por seis dosis. El metotrexato y leucovorín se administraron cada 15 días en siete ocasiones. Además recibió interferón-alfa 4.5 x 10 6 U por vía subcutánea cada tercer día durante las 12 semanas del tratamiento. Resultados. El tratamiento fue bien tolerado alcanzando una respuesta clínica del 70 por ciento en el tumor primario y completa en los ganglios inguinales. Fue sometido a penectomía radical en abril de 1994 y actualmente está libre de enfermedad a los 61 meses de seguimiento. Conclusiones. Esta modalidad de quimioinmunoterapia parece ser muy activa en el cáncer de pene y debería evaluarse en un número mayor de pacientes con el fin de preservar el órgano e incrementar la supervivencia

Persistence of human papillomavirus DNA in cervical lesions after treatment with diathermic large loop excision.

OBJECTIVE: The aim of this study was to identify human papillomavirus (HPV) in cervical intraepithelial neoplasia (CIN) lesions and to evaluate the persistence of viral DNA after diathermic large loop excision (DLLE) treatment. STUDY DESIGN: Biopsies from 36 patients with low- and high-grade CIN lesions were studied before and after DLLE treatment looking for HPV sequences. DNA was extracted to perform a radioactive polymerase chain reaction (PCR) using GP 5,6 generic primers. PCR products were analyzed by the single-stranded conformational polymorphism (SSCP) which is a simultaneous detection and typing method. Dot-blot hybridization with generic and type-specific biotinylated oligonucleotide probes was applied in some cases. RESULTS: HPV DNA was found in all pretreatment samples, and the viral type was identified in 80% of them, HPV 16 being the most prevalent. The viral type coincided with that detected in the first biopsy in all except one case. Seventy five percent of the patients (27 cases) were negative for CIN at follow up, but 50% of them remained HPV DNA positive. CONCLUSION: DLLE treatment was effective in removing the CIN lesion but not the HPV. This fact points out the need to asses the presence of HPV in DNA during the follow-up, since viral persistence has been considered a high risk factor for recurrence and/or malignant transformation.

[Molecular variants of human papillomaviruses types 16, 18, and 45 in tumors of the uterine cervix in Mexico]. / Las variantes moleculares de papiloma virus humanos tipo 16, 18 y 45 en tumores del cuello uterino, en México.

Carcinomas of the uterine cervix still constitutes the first cause of death from cancer among Mexican women. Certain types of human papillomaviruses (HPV) have been implicated in cervical cancer development; active viral sequences are usually found in more than 90% of cervical tumors, their genome contains two oncogenes that immortalize human cells in culture. Recent worldwide studies have shown the existence of molecular variants of known HPV types, mainly 16 and 18, thus permitting the establishment of viral spread during evolution, which seems as ancient as humankind. Phylogenetic studies have identified five major branches for HPV-16 and indicated that viral diversity seems associated with ethnic characteristics of the populations. In this work we searched for the presence of viral sequences among cervical tumors from the Mexican population. The existence of variants of HPV types 16, 18, and 45 was observed. One variant was found in more than half of HPV-16 positive tumors, and seems to exhibit a more aggressive behavior. In HPV-18 positive tumors, in addition to the prototype, two variants were detected in near a fourth of the samples. Finally, all HPV-45 positive tumors showed a new variant not yet reported in the literature. Some of these variants were found associated with specific histological types of cervical cancer, suggesting the participation of these variants in its genesis or aggressivity.

Multimodal tests of cerebrovascular reactivity in migraine: a transcranial Doppler study.

Altered cerebral vasoreactivity (CVR) has been implicated in migraine. To test this hypothesis, we studied CVR as measured by transcranial Doppler ultrasound (TCD) in 11 migraineurs and 12 healthy controls of similar age. Mean flow velocities (MFV) in the middle cerebral artery (MCA) were recorded during a cognitive and two motor tasks. MFV in the posterior cerebral artery (PCA) were measured during photic stimulation and observation of complex images. The increase of MFV in the MCA during the cognitive task was greater in migraineurs than in controls (9.1% vs 5.0%; P = 0.06). The increase of MFV in both tests for PCA reactivity was significantly greater in migraineurs than in controls: 17.4% vs 9.9% for photic stimulation (P less than 0.05) and 20.3% vs 10.2% for observation of complex images (P less than 0.05). Owing to overlap of individual results, the discriminative value of both tests was unsatisfactory. The variability of flow velocities as measured by standard deviations of MFV was significantly greater in migraineurs than in controls during all tests of PCA vasoreactivity. Differences in CVR between migraineurs and normal controls may be detected by TCD testing, in particular in the PCA territory. For individual diagnostic purposes, CVR tests proved to be insufficient.

The amino-terminal half of rotavirus SA114fM VP4 protein contains a hemagglutination domain and primes for neutralizing antibodies to the virus.

We have previously reported the synthesis in Escherichia coli of polypeptide MS2-VP8', which contains the amino-terminal half of the SA114fM VP4 protein fused to MS2 bacteriophage polymerase sequences (C. F. Arias, M. Lizano, and S. López, J. Gen. Virol. 68:633-642, 1987). In this work we have synthesized the carboxy-terminal half of the VP4 protein also fused to the MS2 polymerase. This protein, designated MS2-VP5', was recognized by sera to the complete virion and was able to induce antibodies to the virus when administered to mice; however, these antibodies had no neutralizing activity. The two chimeric polypeptides were tested for their ability to agglutinate erythrocytes and to prime the immune system of mice. Bacterial lysates enriched for the MS2-VP8' hybrid polypeptide, but not those enriched for the MS2-VP5' protein or those containing proteins from the host E. coli strain, had hemagglutinating activity. This hemagglutination was inhibited by sera to SA114fM rotavirus. In addition, a single dose of the MS2-VP8' polypeptide was able to prime the immune system of mice for an augmented neutralizing antibody response when the animals were subsequently immunized with purified SA114fM virus.

Synthesis in Escherichia coli and immunological characterization of a polypeptide containing the cleavage sites associated with trypsin enhancement of rotavirus SA11 infectivity.

About 45% of the rotavirus SA11 VP3 gene was inserted into a thermoinducible expression plasmid under the control of phage lambda PL promoter. The primary translation product predicted on the basis of the plasmid construction was a hybrid protein in which the 98 amino-terminal amino acids of phage MS2 polymerase were followed by amino acids 42 to 387 of the VP3 protein, which included the region containing the cleavage sites associated with trypsin enhancement of infectivity. On induction, a polypeptide that had the expected mol. wt. and contained VP3-related amino acid sequences as judged by immunological criteria, was synthesized to a level representing about 15% of the total bacterial protein. When a bacterial lysate enriched for the fusion polypeptide was injected into mice, it induced antibodies which inhibited haemagglutination and neutralized SA11 rotavirus infectivity.