PubMed Health⌕ Search

Biomedical subjects

S Binda

Publications and source records attributed to S Binda.

At least 19 recordsLinked to original sources
PubMed2003

Retrospective diagnosis of congenital cytomegalovirus infection and cortical maldevelopment.

Congenital cytomegalovirus (CMV) infection can cause malformations of cortical development (MCD). It is difficult to establish CMV as a cause of MCD several months postpartum. This can now be done by detection of CMV DNA in dried blood spots (DBS test) on Guthrie cards. The authors used DBS tests to assess 10 patients with MCD of unknown cause. Four of the 10 patients were positive for CMV.

Cerebral Cortex↗
PubMed2001

CMV gB genotypes and outcome of vertical transmission: study on dried blood spots of congenitally infected babies.

BACKGROUND: The role of the virulence of the infecting cytomegalovirus (CMV) strain in the transmission of the virus from mother to fetus and the outcome of the fetal infection has not received much attention yet. Molecular analysis of the gene coding for the surface glycoprotein B (gB) has been used to investigate the relationship between genotype and virulence in groups of immunosuppressed patients. OBJECTIVES: (1) to assess the prevalence of different gB genotypes in babies with congenital CMV infection; (2) to investigate the possible relationship between genotype and severity of congenital CMV disease; (3) to evaluate the possibility of using dried blood on Guthrie cards (DBS) for genotyping. STUDY DESIGN: CMV DNA was extracted from DBS and from urine/saliva samples collected in the first two weeks of life of 98 congenitally infected babies, half of which were symptomatic at birth. Genotyping was performed through RFLP analysis of the region corresponding to the cleavage site of the gB protein. RESULTS: The most prevalent genotype was gB1 (42%) followed by gB3 (26%), gB2 (19%) and gB4 (13%). Rates of disease and CNS damages were higher among children infected by gB1 (35%, 17%) and gB3 (31%, 28%) than in those infected by gB2 and gB4 (20%, 17% and 13%, 15%, respectively). These differences however did not reach the statistical significance. The parallel typing of DBS and urine/saliva strains gave a full concordance of results. CONCLUSIONS: All four major CMV gB genotypes (gB1-4) can cause a congenital infection but none seems to be associated to the development and the severity of disease. The possibility of using the neonatal DBS for genotyping opens a way to the examination of large numbers of cases of congenital CMV infection.

Blood Specimen Collection↗
PubMed2001

Value of the dorsal cutaneous guinea pig model in selecting topical antiviral formulations for the treatment of recurrent herpes simplex type 1 disease.

Recurrent herpes simplex labialis represents a disease still difficult to treat, despite the availability of many established antiviral drugs used in clinical research since 30 years ago. Although differences between the human disease and that obtained in experimental animal suggest caution in predicting an effective clinical response from the experimental results, some of the animal models seem to be useful in optimising the topical formulation of single antiviral drugs. In the present work the dorsal cutaneous guinea pig model was used to compare 5 different topical antiviral formulations with clinical promise (active molecule: 5% w/w micronized aciclovir, CAS 59277-89-3), using both roll-on and lipstick application systems. The aim being to evaluate which vehicle (water, oil, low melting and high melting fatty base) and application system (roll-on, lipstick) enhances the skin penetration and the antiviral activity of the drug, after an experimental intradermal infection with Herpes simplex virus type 1 (HSV-1). As reference, a commercial formulation (5% aciclovir ointment) was used. The cumulative results of this study showed that the formulation A, containing 5% aciclovir in an aqueous base in a roll-on application system, has the better antiviral efficacy in reducing the severity of cutaneous lesions and the viral titer; among the lipsticks preparations, the formulation D, containing 5% aciclovir in a low melting fatty base, demonstrates a very strong antiviral activity, though slightly less than formulation A. This experimental work confirms the validity of the dorsal cutaneous guinea pig model as a rapid and efficient method to compare the antiviral efficacy of new formulations, with clinical promise, to optimise the topical formulation of the active antiviral drugs.

Acyclovir↗
PubMed2000

Cytomegalovirus DNA detection in Guthrie cards: a powerful tool for diagnosing congenital infection.

BACKGROUND: A simple and reliable diagnosis of congenital cytomegalovirus infection is necessary both for clinical and epidemiological purposes. This could be accomplished through the demonstration of cytomegalovirus (CMV) DNA in blood spots (DBS) on Guthrie cards. OBJECTIVES: (1) To assess the sensitivity and specificity of the method (DBS test) in diagnosing congenital CMV infection compared with viral isolation and (2) to evaluate the applications of the test to the late diagnosis of congenital CMV. STUDY DESIGN: The method was tested on the cards of (1) 509 babies examined through viral isolation within their third week of life (72 positive cases) and (2) 191 children studied after 3 weeks of life (25 days to 5 years). Blood was eluted from Guthrie cards and heat extracted. The products of a nested polymerase chain reaction (PCR) amplifying one region in the CMV glycoprotein B (gB) gene were detected by agarose gel electrophoresis. RESULTS: DBS test was positive in all 72 congenitally infected babies and in four of the 437 negative at cytomegalovirus isolation (sensitivity 100%, specificity 99%). Infection in 16 of the 92 infants with a late viral isolation was demonstrated to be congenital by the test, which also detected congenital infection in 18 of 83 children in whom viral culture was not performed (13 with and five without symptoms). Fifty-six additional control cases tested negative. CONCLUSIONS: DBS test is a reliable assay for diagnosing congenital cytomegalovirus infection and could be used as an alternative to viral culture. It is able to reveal whether ascertained CMV infection is congenital or postnatal at an age when viral isolation is not able to do so. It can assess the role of risky procedures such as transfusion and it can ascertain the etiology of morbid conditions diagnosed late or of controversial origin.

Blood Specimen Collection↗
PubMed2000

Swiss Caucasian population data for the STR loci D2S1338 and D19S433 using the AmpFISTR SGM plus PCR amplification kit.

Allele and genotype frequencies for the ten STR loci D3S1358, VWA, D16S539, D2S1338, D8S1179, D21S11, D18S51, D19S433, TH01, FGA were determined in a Swiss Caucasian population sample (n=206) using the AmpFISTR SGM Plus Amplification kit. Electrophoresis was carried out on an ABI PRISM CE 310 Genetic Analyzer instrument. Previously, allele frequencies were published for the 13 STR loci D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, THO1, TPOX, CSF1PO and D16S539 for the same samples (n=206) amplified with the AmpFISTR Profiler Plus and Cofiler PCR Amplification kits. Since the results for the eight loci D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, THO1, D16S539 shared between the AmpFISTR SGM Plus, Profiler Plus and Cofiler PCR Amplification kits already are published, only the allele frequencies for the two STR loci D2S1338 and D19S433 are reported in this paper. The two loci meet Hardy-Weinberg expectations. In addition, there is little evidence for association of alleles among the 15 loci (amplified with the Profiler, Cofiler, and SGM Plus amplification kits). The allelic frequency data can be used in forensic analyses to estimate the frequency of a multiple STR locus DNA profile in the Swiss population.

Alleles↗
PubMed2000

Phenotypic variability in Bartter syndrome type I.

Limited phenotypic variability has been reported in patients with Bartter syndrome type I, with mutations in the Na-K-2Cl cotransporter gene (BSC). The diagnosis of this hereditary renal tubular disorder is usually made in the antenatal-neonatal period, due to the presence of polyhydramnios, premature delivery, hypokalemia, metabolic alkalosis, hypercalciuria, and nephrocalcinosis. Among nine children with hypercalciuria and nephrocalcinosis, we identified new mutations consistent with a loss of function of the mutant allele of the BSC gene in five. Three of the five cases with BSC gene mutations were unusual due to the absence of hypokalemia and metabolic alkalosis in the first years of life. The diagnosis of incomplete distal renal tubular acidosis was considered before molecular evaluation. Three additional patients with hypokalemia and hypercalciuria, but without nephrocalcinosis in the first two and with metabolic acidosis instead of alkalosis in the third, were studied. Two demonstrated the same missense mutation A555T in the BSC gene as one patient of the previous group, suggesting a single common ancestor. The third patient presented with severe hypernatremia and hyperchloremia for about 2 months, and a diagnosis of nephrogenic diabetes insipidus was hypothesized until the diagnosis of Bartter syndrome type I was established by molecular evaluation. We conclude that in some patients with Bartter syndrome type I, hypokalemia and/or metabolic alkalosis may be absent in the first years of life and persistent metabolic acidosis or hypernatremia and hyperchloremia may also be present. Molecular evaluation can definitely establish the diagnosis of atypical cases of this complex hereditary tubular disorder, which, in our experience, may exhibit phenotypic variability.

Amino Acid Sequence↗
PubMed1999

Bloodstream infections in children with cancer: a multicentre surveillance study of the Italian Association of Paediatric Haematology and Oncology. Supportive Therapy Group-Infectious Diseases Section.

A one-year prospective, multicentre surveillance study on aetiology, main clinical features and outcome of bloodstream infections in children with cancer was conducted in 18 paediatric haematology centres belonging to the Italian Association for Paediatric Haematology and Oncology. A total of 191 bloodstream infections were reported during the study period. Of them, 123 (64%) occurred in neutropenic and 68 (36%) in non-neutropenic patients. Gram-positive cocci caused 45% (85/191) of the episodes, gram-negative rods 41% (78/191), and fungi 9% (18/191). The remaining 5% (10/191) of the episodes were poly-microbial infections. A total of 204 pathogens were isolated (46% gram-positive cocci; 44% gram-negative rods; and 10% fungi). The aetiologic distribution was similar among neutropenic and non-neutropenic patients. A correlation between the infection and the presence of an indwelling central venous catheter was found in 20% (23/114) of the episodes among neutropenic patients and in 55% (23/62) among non-neutropenic patients. Gram-negative micro-organisms were isolated in an unusually high proportion of catheter-related infections (48%). The overall mortality rate from any cause within 30 days from the first positive blood culture was 11%, and was higher among patients who were neutropenic at the onset of the infection than among those who were not neutropenic (15 versus 4%, P = 0.03). In addition, the mortality was significantly higher in recipients of bone marrow transplantation than in patients with acute leukaemia or solid tumour (21, 11 and 6%, respectively) and was also higher in fungaemias and poly-microbial infections (22 and 30%) than in single gram-positive and gram-negative bacteraemias (11 and 6%).

Bacteremia↗
PubMed1998

Congenital cytomegalovirus infection in a northern Italian region. NEOCMV Group.

Knowledge of the prevalence of congenital cytomegalovirus infection is necessary to evaluate the need for prevention. We performed a multicentre one-year study involving 11 neonatology divisions to ascertain the prevalence in Lombardy. Cytomegalovirus was isolated by culturing saliva samples from all babies born (n = 1268) of two 15-day sample periods and from 185 neonates with suspected congenital CMV based on clinical and laboratory findings and the history. The overall prevalence of congenital infection was 0.47% (6/1268) in the sample period group and 5% (9/185) in the second group. Clinical monitoring revealed sequelae in two of three children with symptomatic infection and no asymptomatic child at age two years. In a subgroup of 205 babies including 14 of the infected infants we also evaluated a test to detect cytomegalovirus DNA in the Guthrie cards obtained in neonatal screening for genetic and metabolic disorders. The test's sensitivity was 100% and specificity 98.5%, encouraging its use for early identification of infected neonates and for large epidemiological studies.

Child, Preschool↗
PubMed1998

In vitro activity of acetylsalicylic acid on replication of varicella-zoster virus.

Topical application of a mixture of acetylsalicylic acid (ASA) and diethyl ether is effective in the treatment of acute herpes zoster and postherpetic neuralgia. To study whether the other-than-analgesic effects of that treatment could be due to an antiviral activity of ASA the effects of the drug on the replication of varicella zoster virus (VZV) were assessed by the fluorescent focus assay on MRC5 and Vero cells. ASA caused a marked reduction in the spread of infection in MRC5 monolayers while in growing Vero cells the effective dose proved toxic. ASA concentrations (5-10 mM) which were effective in vitro against VZV are higher than the plasma concentrations attained in the standard treatment of chronic inflammatory states, but are consistent with the skin concentration attained by topical application of ASA/diethyl ether mixture. These data support similar findings relating the antiviral activity of acetylsalicylic acid to influenza virus, CMV, and HIV.

Animals↗
PubMed1997

Fatal overdose of clozapine.

An ingestion of an unknown quantity of Leponex (clozapine) tablets in a suicide is described. Although clozapine is known for over 30 years now, relatively few cases of intoxications due to clozapine overdose have been reported. The authors report a new and quick method to analyze and determine the clozapine and N-desmethylclozapine concentration in body fluids. The analytes and an internal standard (zolpidem) were extracted from alkalinized samples into ethyl acetate before GC/NPD analysis. The proposed method resulted in a rapid procedure most useful in cases of deliberate poisoning with the neuroleptic drug Leponex.

Adolescent↗
PubMed1997

Effects of toluidine blue and destaining reagents used in sexual assault examinations on the ability to obtain DNA profiles from postcoital vaginal swabs.

Toluidine blue is an important tool to detect and document genital and perianal injuries following sexual assault. Application of toluidine blue dye and its subsequent removal from unstained areas by means of a destaining reagent, such as diluted acetic acid or a lubricant has been shown to increase the detection rate of posterior fourchette lacerations from 16% to 40% in adult rape victims. Currently, limited information on toluidine blue positive findings in sexually active control groups imposes some limitation on the interpretation of these injuries. Because injuries could otherwise be attributed to improper handling of an examination speculum or the improper insertion of the examining finger, the toluidine blue test should be performed prior to any digital or speculum examination and thus prior to the collection of forensic evidence. For forensic DNA identity testing, it becomes pertinent to determine whether toluidine blue and the destaining reagents used in a sexual assault examination have an adverse effect on the recovery of high molecular weight DNA from postcoital vaginal swabs and thereby have an impact on restriction fragment length polymorphism (RFLP) analysis or PCR-based tests. It is known that some of the lubricants used can have a destructive effect on sperm motility. In order to investigate the potential effects, postcoital vaginal swabs were taken 6 h after sexual intercourse and exposed directly to 1% toluidine blue in aqueous solution, 1-10% acetic acid, and various surgical and vaginal lubricants. Subsequently, the DNA was isolated and DNA identity typing (RFLP and PCR-based) was performed. The results demonstrate, that these reagents have no negative effect on the ability to obtain DNA profiles, either RFLP or PCR-based, from shallow and deep vaginal swabs. The quantity and quality of extractable high molecular weight DNA obtained was comparable with that from uncontaminated postcoital vaginal swabs. RFLP patterns and PCR-based typing results on the D1S80, HUMTH01, TPOX, and CSF1PO loci were consistent with the uncontaminated control swabs and the corresponding whole blood samples of the donors. Therefore, evidentiary material inadvertently contaminated with these reagents can be successfully typed.

Adult↗
PubMed1997

Persistent high MR signal of the posterior pituitary gland in central diabetes insipidus.

We describe three cases of central diabetes insipidus, each with a different pathogenesis, in which unexpected hyperintensity of the posterior pituitary gland was seen on T1-weighted MR images obtained at the time of presentation. In the first case (idiopathic), the posterior pituitary signal persisted more than 10 years; in the second case (Langerhans cell histiocytosis), the signal disappeared within 3 months, despite early specific chemotherapy with etoposide; and in the third case (transient), the posterior signal disappeared within 1 year, but it was documented at the time of spontaneous reversal of polyuria and polydipsia.

Adolescent↗
PubMed1996

Diagnosis of congenital cytomegalovirus infection by detection of viral DNA in dried blood spots.

BACKGROUND: The reference method of cytomegalovirus (CMV) isolation from urine or saliva is not a feasible routine technique for all newborns, and laboratory diagnosis of this infection would be useful both for epidemiological purposes and to enable prompt institution of adequate measures to identify and correct late sequelae. Extraction and amplification of viral DNA from dried blood spots (DBS) collected from babies in the first days of life during routine screening for genetic and metabolic disorders has been proposed for the early diagnosis of viral congenital infections. OBJECTIVES: To test the method for CMV DNA extraction from DBS and to evaluate the results obtained in newborns with and without a diagnosis of congenital infection based on viral isolation from urine and or saliva at birth. STUDY DESIGN: DBS from Guthrie cards collected in babies who underwent virological tests for CMV infection were tested for CMV DNA by observers blinded to the virological results. DNA was extracted from DBS both in water and in cell culture medium according to Shibata et al. with minor modifications. The products of nested polymerase chain reactions (PCR) amplifying two regions in the IE1 and gp58 genes were detected by agarose gel electrophoresis. Strict control measures were adopted to avoid carryovers and contaminations. RESULTS: DBS from the eight symptomatic and 11 asymptomatic congenitally infected babies were positive when extraction was performed in medium, whereas extraction in water failed to identify two of the asymptomatic cases. The results obtained with the two extraction methods agreed in the remaining cases; the 71 CMV negative control babies were negative and two out of 21 cases of supposed postnatal infection were diagnosed as congenital on the basis of a positive DBS. All positive cases were identified by gp58 PCR but only slightly over half of them by IE1 PCR. Extraction in medium was more efficient than in water. CONCLUSIONS: The method of CMV DNA extraction in medium followed by amplification of the gp58 region showed 100% sensitivity and specificity compared with isolation in cell culture. Therefore, we propose this procedure to diagnose congenital CMV infection at birth and also later.

Journal Article↗
PubMed1996

Cytomegalovirus in peripheral blood leukocytes of infants with congenital or postnatal infection.

BACKGROUND: Cytomegalovirus (CMV) is the most frequent agent of viral infection in the fetus; it causes varying damage, particularly neurologic, which becomes evident at birth or in infancy in about 20% of infected individuals. Postnatal acquisition is usually asymptomatic and without sequelae. Laboratory diagnosis of congenital and postnatal infection is based on the demonstration of virus in urine. OBJECTIVES: To investigate the systemic spread of CMV in neonates with congenital or postnatal infection and to evaluate its significance in diagnosis and in monitoring anti-CMV treatments. DESIGN: Quantitative determinations of infective CMV (viremia) and viral antigen pp65 (antigenemia) were performed on peripheral blood leukocytes (PBL) from the buffy coat of heparinized blood from children with a diagnosis of congenital (n = 19) or postnatal (n = 19) infection based on viral isolation from urine. RESULTS: Antigen pp65 in PBL was detected particularly in children with symptomatic infection, both congenital (100%) and postnatal (79%; P > 0.05), and significantly less frequently (50%; P < 0.001) in those with asymptomatic infection. Viremia was observed less often but always in association with antigenemia. Both tests became negative within 6 months. Neither viral titer nor persistent positivity was related to clinical manifestations. In the nine infants given anti-CMV therapy (ganciclovir and/or hyperimmune gamma-globulins) an early suspension of treatment resulted in the appearance of antigenemia and/or viremia. CONCLUSIONS: Cytomegalovirus was detected in PBL mainly in the most severely affected children. Monitoring antigenemia and viremia in CMV-infected infants is recommended to demonstrate persistent systemic infection and to evaluate virologic results of treatment.

Antigens, Viral↗
PubMed1992

Immunization in children with HIV seropositivity at birth: antibody response to polio vaccine and tetanus toxoid.

OBJECTIVE: To evaluate the humoral response to routine childhood immunization of HIV-infected children. DESIGN: Response rate, antibody titres and persistence after polio and tetanus vaccination were compared in 72 children with HIV seropositivity at birth and divided according to HIV infection status as determined by clinical and laboratory tests. METHODS: Polio antibodies were titred in a microneutralization test (positive titres, > or = 1:4), and antibody to tetanus toxoid with a passive haemagglutination method (protective titres, > or = 1:1024). RESULTS: The response rates to polio and tetanus vaccination (> 80 and > 75%) were similar in the HIV-infected and non-infected children, as were antibody levels. In the subgroup with sera obtained some months after the last dose of vaccine, polio antibody levels decreased in all four HIV-infected and in three of the seven non-infected children; protective tetanus antitoxin levels were detected in three of the six infected and in all three non-infected children. CONCLUSIONS: This study demonstrates the ability of children with HIV infection to respond adequately to the two vaccines considered, although tetanus antitoxin levels were inferior, compared with those in the seroreverted children. The unsatisfactory antibody levels observed in the admittedly few HIV-positive children studied some months after the last vaccination could be the result of a lower initial protective level and not necessarily an expression of severely impaired immunocompetence. The administration of booster doses in addition to the traditional immunization schedule could be useful in children with HIV infection.

Antibodies, Bacterial↗