S. Melón

Minimally invasive treatment of the nasal inverted papilloma.

Background The purpose of this work is to evaluate our results in the treatment of the nasal inverted papillomas with an endoscopic approach using a retrospective case series. Methods Between 1993 and 2000 we treated 27 patients with nasal inverted papillomas. All patients underwent endoscopic nasal surgery under general anesthesia. None of the inverted papillomas extended outside of the paranasal sinuses. All tissue samples underwent polymerase chain reaction and hybridization in situ to detect genetic sequences of the human papilloma virus and Epstein Barr virus. Results The study population consisted of 16 men and 11 women with a median age of 52 years (range, 22–77 years). Ten patients (37%) had undergone a previous nasal surgery. The median follow-up was 5 years (range, 2–8 years). None of the patients presented with bilateral nasal involvement or a synchronous carcinoma. Seven patients underwent an additional surgical approach (two endoscopic approaches via a Caldwel-Luc approach, four sublabial approaches via a Caldwel-Luc approach, and one external ethmoidectomy). There were no surgical complications. Two patients (7%) had recurrent papilloma 4 and 6 years after surgery and again underwent endoscopic resection. The amplification both by polymerase chain reaction and hybridization in situ for human papilloma virus and Epstein Barr virus were negative in the specimens from all patients. Conclusions According to the literature and our own experience, we believe that the initial surgical management of primary and recurrent inverted papillomas limited to the nasal cavity and paranasal sinuses should be endoscopic sinus surgery.

Association between human herpesvirus type 6 and type 7, and cytomegalovirus disease in heart transplant recipients.

CYTOMEGALOVIRUS (CMV) is an important opportunistic infection after heart transplant. The new human herpesvirus, types 6 and 7 (HHV6, HHV7) are two lymphotropic herpesviruses, which, like CMV, have the potential to be pathogenic in immunocompromized individuals. Recent studies suggest that both viruses can cause clinical illness after transplantation. These viruses may be a risk factor for CMV disease in posttransplant patients, possibly through a direct interaction or through a general immunomodulatory effect. However, the pathogenic potential and the clinical features have not been defined. To evaluate the reactivation of HHV6 and HHV7 after heart transplantation and their implication in the cytomegalovirus disease (CMVD), 42 unselected heart transplant patients were studied.

Diagnosis of herpetic keratoconjunctivitis by nested polymerase chain reaction in human tear film

A study was performed to evaluate nested PCR (nPCR) versus viral cultures as method and tear film versus corneal scrapings as specimen in the diagnosis of viral keratoconjunctivitis. Tear film specimens were taken from both eyes and corneal scrapings from the affected eye only in 17 patients with suspected viral keratoconjunctivitis. In 15 of the 17 patients the viral agent of the infection could be detected: 11 patients had herpes simplex virus type 1, two varicella-zoster virus, one both herpes simplex Virus type 1 and varicella-zoster virus, and one adenovirus. Overall there was no significant difference between the detection rate for corneal scrapings (85%) and tear film (75%). In both types of specimens nPCR showed a higher detection rate than viral cultures (corneal scrapings: 87.5% vs 31.25%; tear film: 75% vs 12.5%;P 0.05). For the diagnosis of keratoconjunctivitis nPCR is superior to viral culture and tear film is an adequate sample that is easier to collect, causing the patient less discomfort.

Detection of human bocavirus in Asturias, Northern Spain

Viral respiratory tract infections cause a substantial amount of illness and death in children worldwide. The so-called respiratory viruses that include respiratory syncytial virus (RSV), influenza virus A (IA) and influenza virus B (IB), human parainfluenza viruses (PIV), adenovirus, coronavirus, rhinovirus, the recently discovered human metapneumovirus (hMPV), and several coronaviruses (SARS, NL63, HKu1) likely account for a majority of respiratory tract illnesses. However, in a substantial proportion of respiratory tract infections, no etiologic agent is detected, even when sensitive detection methods, such as a polymerase chain reaction (PCR), are used. This finding suggests that unidentified pathogens may be circulating and causing disease in children. Recently, a new member of the Parvoviridae family, the human bocavirus (HBoV), has been cloned from nasopharyngeal aspirates of children with acute respiratory infection (ARI) in Sweden [1]. In spite of the few reports published, HBoV appears to be very common. Thus, it has been detected in 1.5%–11.3% of individuals with ARI in Europe, the United States, Canada, Asia, South Africa, and Australia. The clinical manifestation includes cough, fever, rhinitis, nasal congestion, myalgia, respiratory difficulty, bronchiolitis, and pneumonia [1–10]. The aim of this study was to detect HBoV prospectively in samples obtained from children who visited the Pediatric Emergency Service, and to evaluate its relative contribution as a cause of different respiratory infections with respect to other respiratory viruses. From 20th April to 24th November 2006, 366 samples (209 pharyngeal, 148 nasal, five nasopharyngeal swabs, two bronchial aspirates, and two bronchoalveolar lavage fluids) from 339 children (190 males and 149 females) with a mean age of 2.9±3.4 years, (range 3 days–13 years) and with ARI were collected for viral diagnostic evaluation. Of them, 118 (34.8%) showed symptoms of lower respiratory tract infections (LRTI), such as bronchiolitis and pneumonia, 115 (33.9%) came to hospital with symptoms of upper respiratory tract infections (URTI), such us pharyngitis, laryngitis, or rhinitis, and the remaining 106 (31.3%) showed only general symptoms of respiratory infection (SRI), such as fever, myalgia, headache, or cough. The samples were tested with monoclonal antibodies to RSV; IA and IB viruses; PIV type 1, 2, and 3; and human adenovirus. The samples were also inoculated into MRC-5, LLC-MK2, and MDCK cell monolayers following standard protocols. Also, they were processed for rapid cultures (“shell-vial”) in MRC-5 monolayer cells for above mentioned viruses (RSV, IA, IB, PIV, and adenovirus). Viral genomes were purified by using an automated nucleic acid purifier (AmpliPrep, Roche Diagnostics, USA). The routine detection of respiratory viruses comprised nested reverse transcription polymerase chain reaction (RT-PCR) for RSV, IA, IB, IC, PIV, hMPV, and coronaviruses. In 106 pharyngeal swabs, a PCR to detect the Epstein-Barr virus (EBV) was also performed. Furthermore, an “in-house” nested PCR with primers deduced from the NP-1 gene of HBoV was developed. The first round of amplification was carried out with the outer Eur J Clin Microbiol Infect Dis (2008) 27:237–239 DOI 10.1007/s10096-007-0419-9

Involvement of adenovirus in clinical mononucleosis-like syndromes in young children

Although Epstein-Barr virus (EBV) commonly causes infectious mononucleosis (IM) or IM-like syndromes, other agents can be implicated. In this study, viral and parasitic screening was performed to determine the etiological agent of pediatric IM-like syndromes in 38 children. Adenovirus was the agent most frequently detected (47.3%), followed by EBV (31.5%) and cytomegalovirus (2.6%). Although the statistically significant difference between viral detection rates observed in patients who fulfilled clinical and hematological criteria and detection rates in those who presented clinical symptoms only (91.6% vs. 64.3%) indicates that hematological abnormalities are common in viral IM-like syndromes, the existence of syndromes of viral etiology without hematological criteria cannot be discarded. A further analysis showed an absence of lymphocytosis in adenovirus infections as well as a low number (14.3%) of EBV infections associated with increased neutrophils. These data suggest the usefulness of appropriate virological techniques for the detection of adenovirus in pediatric IM-like syndromes.

Molecular identification of two genotypes of mumps virus causing two regional outbreaks in Asturias, Spain

BACKGROUNDnIn spite of universal vaccination, several sporadic cases of mumps infection, which could produce outbreaks, are detected every year in different countries.nnnOBJECTIVEnMumps virus strains causing two regional outbreaks in Asturias (Spain) were phylogenetically characterized.nnnSTUDY DESIGNnMumps virus strains, which were detected in samples from patients belonging to two regional outbreaks in Asturias, were characterized by sequencing of the SH gene and further alignment to homologous sequences of representative strains of the different mumps genotypes.nnnRESULTSnTwo different strains (Ast/SP02 and Ast/SP07) were isolated. Sequence analysis revealed that while Ast/SP02 belonged to genotype H, Ast/SP07 was phylogenetically close to UK02-19, a reference strain for a new genotype. Both strains belonged to different genotypes from those used in the vaccination (Jeryl-Lynn strain is genotype A).nnnCONCLUSIONnMumps virus strains different from those used in vaccination program can cause mumps outbreaks even in vaccinated patients.

Comparative study of the efficacy of an induction dose of interferon-alpha2b with ribavirin compared with standard combined treatment in naive patients with chronic hepatitis C.

Summary. The efficacy and secondary effects of an induction dose of interferon‐α2b (IFN‐α2b) with ribavirin compared with standard combined treatment in naive patients with chronic hepatitis C infection were compared. A prospective study was undertaken between March 1998 and November 2001 in which 84 Spanish hospitals took part. Six hundred and fourteen naive patients (age range 18–65u2003years) diagnosed with chronic hepatitis C virus (HCV) infection and without cirrhosis or co‐infection by other viruses, were included. Patients were divided into two groups. Group A (nu2003=u2003304) received induction treatment with a daily dose of 5u2003MU of IFN‐α2b for 4u2003weeks, followed by 5u2003MU three times a week with ribavirin (1000–1200u2003mg/day, according to weight) until completing 1u2003year of treatment. Group B (nu2003=u2003310) received the standard dose of IFN‐α2b of 3u2003MU three times per week for 48u2003weeks together with ribavirin (1000–1200u2003mg/day, according to weight). Both groups were completely comparable according to age, gender, body weight, transaminase levels, genotype, viral load and hepatic inflammatory activity (Knodell Index). No control group was included for ethical reasons. Pegylated interferon was not available at the time of the study. Serum baseline samples were collected for the determination of genotype. Samples were also collected at baseline, weeks 4, 12, 24, 48 and 72, in order to detect and quantify HCV‐RNA. The efficacy of treatment was evaluated by means of sustained viral response (SVR) characterized by persistent negativity of HCV‐RNA at the end of the follow‐up period.

The mmp1 (−16071g/2g) single nucleotide polymorphism associates with the Haart-related lipodystrophic syndrome

Objective:Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are involved in extracellular matrix remodelling and adipocyte differentiation and are inhibited by antiretrovirals. MMPs and TIMPs and their single nucleotide polymorphisms (SNPs) might contribute to the HAART-related lipodystrophic syndrome pathogenesis. Design and setting:Cross-sectional study in a university-based outpatient clinic. Patients and methods:Two hundred and sixteen HIV-infected patients on extended HAART were studied. Serum MMPs (1, 2, 3, 8, 9, 10, 13) and TIMPs (1, 2, 4) were measured by ELISA microarrays. MMP1 (−16071G/2G) SNP was also genotyped. Lipodystrophic syndrome was diagnosed by a clinical scale validated by fat dual energy X-ray absorptiometry. Results:Eighty-two patients (38.0%) showed lipodystrophic syndrome, mostly lipoatrophy. The 2G/2G MMP1 SNP genotype was more frequent among lipodystrophic syndrome patients (41.3 vs. 20.5%, odds ratio, 2.73; 95% confidence interval, 1.41–5.29; χ2 = 9.62, P = 0.002 for HIV-infected patients with and without lipodystrophic syndrome respectively). Carriers of this genotype had higher serum levels of MMP1 compared with those with the 1G/1G (P = 0.02). Higher MMP1 (P = 0.022) and lower TIMP4 (P = 0.038) serum levels were observed while comparing HIV patients with and without lipodystrophic syndrome. MMP1 2G carriage (P = 0.0008), TIMP4 lower serum levels (P = 0.02), treatment with stavudine (P < 0.0001), treatment with zidovudine (P = 0.006) and absence of hepatitis C virus coinfection (P = 0.002) were associated with lipodystrophic syndrome by logistic regression. Conclusion:MMP1 SNP, which induced increased serum levels of this protein, was associated with lipodystrophic syndrome.

Amplification of herpes simplex virus type 1 DNA in human geniculate ganglia from formalin-fixed, nonembedded temporal bones

Polymerase chain reaction (PCR) has provided new insights in molecular biology. Recently, some studies have been focused on temporal bone pathology, with amplification of DNA from fixed sections of celloidin-embedded bones. The purpose of our study was to elucidate the utility of PCR in detection of minor concentrations of DNA from nonoptimal stored samples. We obtained geniculate ganglia from 30 temporal bones preserved in formalin for a long time, without any process of embedding. By performing a nested PCR assay, we detected herpes simplex virus type 1 DNA in 13 of 30 ganglia (43%). We conclude therefore that study of temporal bones stored under poor conditions by PCR is possible, although there are some limitations when compared with fresh or optimally archived samples. (Otolaryngol Head Neck Surg 2000;123:508-11.)

Detection of herpes simplex virus-1 by nested PCR. An experimental model

Nested polymerase chain reaction (nested PCR) was performed using a reaction mix batch-prepared and kept frozen in single reaction tubes at -20 degrees C until use. Twenty-one New Zealand white rabbits were infected with herpes simplex virus type 1 (HSV-1). Eleven animals were killed on day seven and the other ten were sacrificed on day 21. Viral culture and nested PCR was used to determine the presence of HSV-1 in samples from the tongue, HSV-1 was detected in 90.47% of the animals; in 84.21% by nested PCR and in 52.63% by culture. Nested PCR assay had greater sensitivity than culture in animals sacrificed on day seven with significative difference (p < 0.05). Higher sensitivity and faster results were obtained with this method, so we found it reliable and useful in the setting of a clinical laboratory dealing with diagnosis of herpes virus infections.