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Biomedical subjects

M Comar

Publications and source records attributed to M Comar.

30 records · Page 2Linked to original sources

Quantitative dynamics of HIV type 1 expression.

A competitive PCR and RT-PCR procedure was developed for the quantification of HIV-1 nucleic acids in infected biological samples, with particular reference to the study of the kinetics of production of differently processed viral transcripts. The procedure entails the utilization of a competitor plasmid DNA (on DNA samples) or of an in vitro transcription product obtained from this plasmid (on RNA samples) and allows the quantification of proviral DNA, viral genomic RNA, and viral single- and multispliced mRNAs. Furthermore, it permits the direct standardization of these measurements to the amount of a reference cellular gene (for DNA quantification) or of a reference cellular transcript (for RNA quantification). This quantification procedure was used to monitor the dynamics of HIV-1 transcriptional activation in the latently infected U1 monocytic cell line after stimulation with phorbol-12-myristate-13-acetate, and in experimentally infected peripheral blood lymphocytes. Despite the biological differences between the two experimental systems, in both cases production of infectious virus is accompanied by a remarkable increase in the levels of unspliced viral mRNAs (rising up to 20,000 fold in U1 cells) and by a consequent switch in the abundance of the differently spliced transcript classes. These observations reinforce the notion that the control of infection is subjected also to posttranscriptional events and prompts for quantitative evaluation of HIV-1 transcript class abundance in infected individuals to define potential markers for disease progression.

Base Sequence↗

Dynamics of viral replication in infants with vertically acquired human immunodeficiency virus type 1 infection.

About one-third of vertically HIV-1 infected infants develop AIDS within the first months of life; the remainder show slower disease progression. We investigated the relationship between the pattern of HIV-1 replication early in life and disease outcome in eleven infected infants sequentially studied from birth. Viral load in cells and plasma was measured by highly sensitive competitive PCR-based methods. Although all infants showed an increase in the indices of viral replication within their first weeks of life, three distinct patterns emerged: (a) a rapid increase in plasma viral RNA and cell-associated proviral DNA during the first 4-6 wk, reaching high steady state levels (> 1,000 HIV-1 copies/10(5) PBMC and > 1,000,000 RNA copies/ml plasma) within 2-3 mo of age; (b) a similar initial rapid increase in viral load, followed by a 2.5-50-fold decline in viral levels; (c) a significantly lower (> 10-fold) viral increase during the first 4-6 wk of age. All infants displaying the first pattern developed early AIDS, while infants with slower clinical progression exhibited the second or third pattern. These findings demonstrate that the pattern of viral replication and clearance in the first 2-3 mo of life is strictly correlated with, and predictive of disease evolution in vertically infected infants.

DNA, Viral↗

Treatment of human immunodeficiency virus infection with hydroxyurea: virologic and clinical evaluation.

To assess the efficacy of the therapeutic use of inhibitors of ribonucleotide reductase for the treatment of human immunodeficiency virus (HIV) type 1 infection, 7 consecutive patients were enrolled in a clinical trial involving monotherapy with hydroxyurea for 8-19 weeks. During therapy, patients were evaluated for clinical status and immunologic, hematologic, and quantitative virologic parameters, including determinations of viremia and the number of provirus-containing cells by competitive polymerase chain reaction. In all patients, these parameters were not modified during the course of therapy. The number of CD4 cells remained generally unchanged or showed a tendency to further decline. No sign of improvement in HIV disease was detected in any patient. These observations indicate that monotherapy with hydroxyurea does not provide therapeutic benefit to HIV-1-infected patients.

Acquired Immunodeficiency Syndrome↗

[Maternal-fetal transmission of human papillomavirus].

AIM: Human papilloma virus (HPV) infection is one of the most frequently observed sexually-transmitted diseases (10-60% of the general population). In pregnant women, as well as accelerating the evolution of dysplasia to cervical cancer, the infection may be transmitted to the fetus during gestation or at the time of birth. Children who have been infected at birth may develop laryngeal papillomatosis during the first 5 years of life that may, in some cases, spread to the point of causing aphonia or severe respiratory obstruction. There is also the risk, although it is very low, of a carcinomatous degeneration of the larynx in these subjects during adulthood. The hypothesis of the present study was to verify the prevalence of HPV infection in a population of pregnant women and the prevalence of maternal-fetal transmission. EXPERIMENTAL DESIGN: A prospective longitudinal design lasting 11 months was used for the study. It included the collection of an endocervical biopsy from population of pregnant women using a swab that was diluted in 3 cc of physiological solution, and the collection of oropharyngeal secretions from their respective neonates using a cottonwool bud. PARTICIPANTS AND METHODS: A total of 170 pregnant women attending the Obstetric Centre of the Obstetric and Gynecological Clinic of Trieste University were recruited in the study. An endocervical biopsy was taken during the 1st and/or 2nd trimester of gestation and/or at the start of labour. Of these subjects, 23 completed all the planned biopsies and a sample of oropharyngeal secretion was collected from their neonates. TESTS: From the material obtained the presence of HPV-DNA was analysed using a PCR (protein chain reaction) technique consisting of the following steps: 1) culture of human cells expressing sequences of HPV 16 and 18 used as positive controls; 2) preservation of tissue material washed in watery 4% formalin solution; 3) amplification and viral characterization in types 6-11-16-18-31-33-52. RESULTS: Positive HPV-DNA results in at least one of the three samples collected during the various periods of gestation was 31.2%, whilst in the population in which all the planned samples were performed the frequency of positive cases was 30.4%. Positive results for HPV-DNA in oropharyngeal secretions from neonates was 21.7%. The concordance of positivity for HPV-DNA in mothers at the time of labour and in their respective neonates was 57.14%. CONCLUSIONS: The trend of infection did not reveal substantial changes during the various gestational periods in which tests were performed. The possibility of HPV-DNA transmission from mother to fetus is high, above all when the maternal PCR test is positive at the time of birth, or in the presence of a high viral load. This justifies the need to monitor this infection in pregnancy in those affected by florid genital condylomatosis or with koilocytosis on cervical cytology. It is also appropriate to check all HPV-DNA positive neonates one year after birth.

Biopsy↗

[Vertical transmission of HGV and outcome of the infected babies].

To assess the risk of HGV mother-to-infant transmission and the clinical outcome of infected babies, we investigated 103 mother-infant couples and followed-up the infected children for 4-72 months. Twenty (19.4%) mothers were HGV-RNA positive and transmission occurred in ten (50%) babies; only one child acquired HGV and HCV infection. Maternal factors, such as history of intravenous drug use, HCV-RNA positivity, HIV coinfection, type of delivery and type of feeding were not related to HGV transmission. One HGV infected baby showed a mild hepatitis when he was also infected by Cytomegalovirus. Two babies cleared HGV within the first year of life. The HGV transmission rate is elevated but HGV infection seems to be benign, at least in a short-term follow-up.

Female↗

[HHV-6, new perspectives].

HHV-6 is the etiological agent of Exanthema subitum, and its role in human infection is well known. Recently, molecular diagnostics tools showed for HHV-6 new pathogenetic features and new clinical implication. The present paper highlights recent knowledge on HHV-6 infection and presents a number of results concerning HHV-6 infection in children who had undergone BMT and concerning the roles of endothelial cells as viral reservoir.

Adult↗

[Intravenous immunoglobulins in bone marrow transplantation].

Intravenous immunoglobulins have a role also in the prevention of infections after a bone marrow transplantation. Even if a reduction of CMV pneumonia is reported it is not clear whether such an effect is due to a direct antiviral activity or it is mediated through an immune modulation that reduces GVHD and then the immune suppression. We are currently using i.v.Ig at a dose of 100 mg/kg/day within a protocol for GVHD prophylaxis after a mismatched BMT.

Bone Marrow Transplantation↗