SCN4A mutation as modifying factor of myotonic dystrophy type 2 phenotype.

In myotonic dystrophy type 2 (DM2), an association has been reported between early and severe myotonia and recessive chloride channel (CLCN1) mutations. No DM2 cases have been described with sodium channel gene (SCN4A) mutations. The aim is to describe a DM2 patient with severe and early onset myotonia and co-occurrence of a novel missense mutation in SNC4A. A 26-year-old patient complaining of hand cramps and difficulty relaxing her hands after activity was evaluated at our department. Neurophysiology and genetic analysis for DM1, DM2, CLCN1 and SCN4A mutations were performed. Genetic testing was positive for DM2 (2650 CCTG repeat) and for a variant c.215C>T (p.Pro72Leu) in the SCN4A gene. The variation affects the cytoplasmic N terminus domain of Nav1.4, where mutations have never been reported. The biophysical properties of the mutant Nav1.4 channels were evaluated by whole-cell voltage-clamp analysis of heterologously expressed mutant channel in tsA201 cells. Electrophysiological studies of the P72L variant showed a hyperpolarizing shift (-5 mV) of the voltage dependence of activation that may increase cell excitability. This case suggests that SCN4A mutations may enhance the myotonic phenotype of DM2 patients and should be screened for atypical cases with severe myotonia.

Quantificação de Mycobacterium leprae em amostras de secreção nasal de casos paucibacilares e multibacilares por PCR quantitativo

A quantificação do bacilo realizada pelo exame baciloscópico e histopatológico apresenta sensibilidade limitada. Portanto, o emprego de uso de técnicas moleculares permite o diagnóstico direto do material clínico com elevada especificidade e sensibilidade. A PCR em tempo real (qPCR) é um ensaio sensível e específico que permite a quantificação do número de bacilos a partir de diversas amostras, além de poder ser utilizada no diagnóstico diferencial de muitos patógenos. Pacientes multibacilares hansenianos liberam o bacilo Mycobacterium leprae através da secreção nasal, sendo a coleta deste realizada por procedimento não invasivo. Até o momento, nenhum estudo avaliou a sensibilidade e especificidade da qPCR para o diagnóstico da hanseníase utilizando amostras de secreção nasal.

Neuropsychological profile of pre-schoolers with metaphonological difficulties: results from a non-clinical sample.

BACKGROUND: The level of language development reached in pre-school age is considered the most reliable predictor of reading acquisition. In normally developing children, learning to read is strongly related to early language skills, and in particular to phonological processing abilities. In dyslexic children, reading abilities seem to show a correlation with phonological awareness. METHODS: A group of 65 children (aged 5-6 years) were recruited and submitted to an in-depth neuropsychological assessment [i.e. metaphonological skills, intelligence, verbal short-term memory (VSTM) and other aspects of receptive and expressive language]. We were able to identify 14 children with significant metaphonological difficulties (MD): 11 children with exclusively MD, and the other three children with specific language impairment. This study compares the neuropsychological profile obtained from children with MD with that of a peer group without any language impairment (N). RESULTS: The performances of the MD were within the normal ability range in almost all the administered tests but significantly lower compared with those of their peers without language impairment (N) in some items of the intelligence scale (Wechsler Preschool and Primary Scale of Intelligence) and in the tests of VSTM and of receptive/expressive language. Nevertheless, there were not statistically significant differences between MD and N in output phonology. CONCLUSIONS: In pre-school age, in a group of non-clinical children, with a range of abilities, those with MD appear to be at the lower end of the normal range in many other verbal skills. These children could be considered at-risk for possible subsequent difficulties learning to read and thus need to be identified and to warrant prompt treatment.

Participation of Na,K-ATPase in FGF-2 secretion: rescue of ouabain-inhibitable FGF-2 secretion by ouabain-resistant Na,K-ATPase alpha subunits.

We have examined the relationship between Na,K-ATPase and FGF-2 secretion in transfected primate cells. FGF-2 lacks a classic hydrophobic export signal, and the mechanisms mediating its secretion are unknown. To monitor secretion, a FLAG epitope tag was inserted into the carboxyl terminus of the 18 kDa form of human FGF-2, and the construct was transfected into either human HEK 293 or monkey CV-1 cells. Exported FGF-2 was detected in the culture medium using the FLAG-specific monoclonal antibody M2. FGF-2 secretion from HEK 293 or CV-1 cells was linear over time and sensitive to inhibition by the cardiac glycoside ouabain, a specific inhibitor of the Na,K-ATPase. In contrast, the secretion of FGF-8 (an FGF family member that contains a hydrophobic secretory signal) was not inhibited by treatment of HEK 293 or CV-1 cells with ouabain. FGF-2 secretion was also assayed in CV-1 cells expressing the naturally ouabain-resistant rodent Na,K-ATPase alpha1 subunit. In cells expressing the rodent alpha1 subunit, FGF-2 secretion was unaffected by high levels of ouabain, indicating that the rodent alpha1 subunit was capable of rescuing ouabain-inhibitable FGF-2 export. Expression of ouabain-resistant mutants of the rodent alpha2 and alpha3 subunits, or the naturally ouabain-resistant rodent alpha4 subunit, also supported FGF-2 secretion in ouabain-treated cells. Taken together, our studies are consistent with the idea that the Na,K-ATPase plays a prominent role in regulating FGF-2 secretion, although none of the alpha subunit isoforms exhibited specificity with regard to FGF-2 export.

Fecal alpha-1-antitrypsin excretion in children with diarrhea.

BACKGROUND: The aim of this study was to compare the concentration and immunoelectrophoretic characterization of alpha-1-antitrypsin (AAT) excreted in random fecal samples in children with acute and chronic diarrhea and in control groups. METHODS: Thirty-two children with diarrhea and 23 healthy children were evaluated. The concentration and characterization of AAT were determined by radial immunodiffusion and crossed immunoelectrophoresis, respectively. RESULTS: The increase in the concentration of fecal AAT was more than the upper limit for the control group (1.25 mg/g of dry stool mass) in the patients with chronic infectious diarrhea and in 52% of those with chronic non-infectious diarrhea but not in those with acute diarrhea, infectious or non-infectious. Immunoelectrophoretic analysis showed two forms of fecal AAT in both sick and healthy children. The alterations in the concentration of fecal AAT did not correlate with the immunoelectrophoretic pattern of AAT. CONCLUSION: Our results suggest that the determination of fecal AAT could give clinically useful information about the difference between infectious and non-infectious diarrhea and the activity of characterizing disease with diarrhea.

[Viral respiratory infections in small children in the season in the season 1996-1997]. / Wystepowanie wirusowych zakazen dróg oddechowych u malych dzieci w sezonie 1996-1997.

Using the immunofluorescence method, the etiology of virus respiratory infection was assayed in children aged 0-2 years, hospitalized because of respiratory diseases in the epidemic season 1996-1997. Data obtained were compared with those from the preceding season (1995-1996), as well as, from the 10 preceding seasons (1985-1995). In the season under study, participation of respiratory viruses was a little higher in the infections to be hospitalized. As compared with the preceding season, and the preceding 10 seasons, infections with parainfluenza type 1 and 3 viruses were higher in statistically significant way. Compared with the preceding seasons, more viral respiratory infections were found in November 96, as well as, in January and February 97. The number of viral respiratory infections was increased among children aged 10-12 months.

Neuropsychological deficits in asymptomatic atrial fibrillation.

OBJECTIVE: To assess the preclinical effects on cognitive functions of nonrheumatic atrial fibrillation (NRAF) in patients with negative history for cerebrovascular disease. MATERIALS AND METHODS: The study included 37 consecutive patients with chronic (n=16, mean age 65.3+/-6.6 years) or paroxysmal (n=21, mean age 58.3+/-9.5 years) NRAF and an equal number of control subjects in sinus rhythm, who were matched for age, education and presence of hypertension. A comprehensive neuropsychological battery including tests of attention, memory, language and visuospatial skills was administered. RESULTS: Patients with chronic NRAF showed significantly poorer performances in tasks exploring attention and verbal memory functions, while the paroxysmal group was significantly impaired in a long-term memory task. The neuropsychological findings were confirmed excluding from both groups patients with CT evidence of cerebrovascular damage. A small subgroup of patients was also submitted to cerebral MRI. CONCLUSION: Neurologically asymptomatic NRAF is related to a subclinical but significant impairment in attention and memory. These deficits could be produced by minor ischemic lesions due to microembolization, or by diffuse hypoxic damage due to hypoperfusion.

Influence of residual ischaemia on heart rate variability after myocardial infarction.

Despite the growing evidence for the positive predictive value of depressed baroreflex sensitivity and/or reduced heart rate variability after myocardial infarction, the mechanisms involved in these autonomic alterations are not fully understood. Specifically, the possible influence of residual ischaemia has not been assessed. To address this problem we studied the spectral analysis of heart rate variability in 21 patients with a first myocardial infarction in whom the only clinical correlate was the presence of residual ischaemia, as documented by the positive response to both an exercise stress test and an echocardiographic stress test. Data from these patients were compared with those obtained in a group of postmyocardial infarction patients similar for several risk factors, age, site of myocardial infarction, but without residual ischaemia. Patients positive for residual ischaemia had lower power in the whole spectrum (1146 +/- 158 vs 1631 +/- 159 ms2, P = 0.032) as well as in the low and high frequency bands of heart rate variability. A nocturnal increase in high frequency was observed in those without residual ischaemia (from 167 +/- 35 to 242 +/- 51 ms2, +45%, P = 0.034), but not in those with residual ischaemia (from 111 +/- 19 to 141 +/- 29 ms2, +27%, ns). Thus, residual ischaemia reduces heart rate variability after myocardial infarction. The autonomic effects of residual ischaemia probably contribute to its negative prognostic value after myocardial infarction.

[Breast feeding for prevention of viral acute respiratory diseases in infants]. / Karmienie naturalne w zapobieganiu wirusowym zakazeniom dróg oddechowych niemowlat.

The incidence of acute viral respiratory diseases (confirmed by immunofluorescence test) was compared in a group of 159 infants aged 0-9 months breast-fed at least once daily and in a group of 608 infants of similar age and hospitalization duration receiving only artificial food. The incidence was significantly lower in breast-fed infants (p = 0.00057), especially in the first trimester of life (p = 0.00039). This was valid of all RSV infections (p = 0.01854) and parainfluenza type 4 virus infections (p = 0.0846). The antibody level against parainfluenza type 1 and 3 (HI) and against RSV in IgA class (ELISA) was compared with that in mother's milk and serum. No significant correlation was found for parainfluenza type 1 antibodies (r = 0.159) and 3 (r = 0.025), while for RSV the correlation was positive and statistically significant though rather low (r = 0.234). No significant correlation was found between milk antibody levels and the incidence of acute viral respiratory infections. Breast-feeding showed a significant influence on the lower incidence of these infections, particularly in the first trimester of life. It seems that this was associated with a non-specific mechanism rather than with the milk levels of specific antibodies.

[Viral respiratory infections detected in small children in 1995-1996]. / Wirusowe zakazenia dróg oddechowych u malych dzieci w sezonie 1995-1996.

As in previous studies we used the fluorescent antibody technique to determine the aetiology of viral respiratory infections in small children who were hospitalized in the epidemic season 1995-96. Data obtained from this study were compared with the results from the previous season (1994-95) and with the results from preceding ten seasons (1985-95). No variation in aetiology of infections was found although a higher occurrence of adenoviruses was uncovered amongst the 1995-1996 group. The monthly rates of morbidity in this same group were highest in November and in April. Data from this season displayed the greatest incidence of morbidity among children from one to six months old. The importance of bread feeding as a measure of prevention against viral respiratory infections was confirmed.

[Interstitial pulmonitis due to the cytomegalovirus in an immunocompetent adolescent: treatment with ganciclovir]. / Polmonite interstiziale da citomegalovirus in adolescente immunocompetente: trattamento con il Ganciclovir.

Interstitial pneumonia is the most common disease caused by infection from cytomegalovirus (CMV) in immunodepressed patients, whereas it is a rare complication in immunocompetent patients. With reference to the second group of patients, little literature has been produced as for the therapy to choose when symptoms are serious. We report the case of immunocompetent adolescent with CMV pneumonitis who responded dramatically to therapy with ganciclovir. For a week B. M., a 15-year-old girl, has been showing fever, cough and boring pain at her left thoracic base. When hospitalized, the girl was suffering and dyspneic, cardio-thoracic conditions were bad. Spleen and liver were palpable two fingers far form costal arch. Hematochemical tests showed an increase in phlogosis and transaminase value. Thoracic X-ray was negative, as well as cultures. Among the serological tests high response of anti-CMV IgM was remarkable. Virological blood test confirmed active CMV infection. On the fifth day, a thoracic radiography showed widespread interstitial infiltrates. Treatment with ganciclovir--i.v. 6 mg/kg/day, twice a day for twelve days--has been then adopted. After two day treatment, the girls was apyretic and eupneic. After ten day treatment, thoracic radiography was negative and a great decrease in CMV antigenic response was given by blood tests. No side effect were observed. According to our experience we can say that treatment with ganciclovir may positively shorten the course of pneumonia caused by CMV in immunocompetent patients.

[The incidence of virus respiratory infections in small children in the season 1994-95]. / Wystepowanie wirusowych zakazen dróg oddechowych u malych dzieci w sezonie 1994-1995.

As previously, by antibody fluorescent technique the etiology of viral respiratory infections was determinated in small children hospitalized because of respiratory disease in the epidemic season 1994-95. Data obtained were compared with the results from the preceding season. Among a total of 371 patients, virus infection was detected in 170 (45,8%). Data for the whole season did not distinctly differ from those obtained previously; however, some differences were found in the occurrence of the individual virus infections in consecutive months of the epidemic season. The second (spring) wave of RS virus infections in this season was lower and more extended in time than in the previous seasons. Highest incidence of adenoviruses was observed in July 1995. The incidence of parainfluenza type 4 virus was higher in the disease of small children. In the youngest (below 1 month) age group percent of virus infections was higher, as compared with data from the preceding season.

Transfusion-transmitted human parvovirus B19 infection in a thalassemic patient.

BACKGROUND: Human parvovirus (HPV) B19 infection has been shown to be transmissible by clotting factor concentrates, most often resulting in asymptomatic seroconversion. So far, no case of B19 transmission due to single-donor transfusion has been documented. CASE REPORT: A case of transfusion-transmitted HPV B19 infection in a 22-year-old female thalassemia major patient is described. She presented with an aplastic crisis; this was followed 1 week later by transitory heart failure and acute tricuspid incompetence. The echocardiogram revealed a grade III tricuspid regurgitation and a floating vegetation on the atrial face of the tricuspid lateral leaflet. The tricuspid regurgitation and vegetation spontaneously disappeared within 15 days. Blood cultures for bacteria were repeatedly negative. IgM anti-HPV B19 seroconversion was documented in the acute phase. B19 DNA was detected by polymerase chain reaction and remained detectable up to 4 months after diagnosis. High-titer IgM anti-HPV and B19 DNA were also found in serum samples collected at the time of donation from one of the donors of the blood transfused before the onset of clinical symptoms. CONCLUSION: This case documents the transmission of HPV B19 by the transfusion of 1 red cell unit and the occurrence of possible transient cardiac involvement in this infectious complication.

[Bacterial flora in the digestive tract during diarrhea in infants upon hospital admission and at the end of hospitalization]. / Bakteryjna flora przewodu pokarmowego w przebiegu biegunki u niemowlat na wstepie i przy koncu hospitalizacji.

The stools of 23 children aged from 14 days to 18 months were analysed (qualitatively and quantitatively) for some genera of bacteria which could be the cause of diarrhea. Feces were collected at the beginning and the end of hospitalisation. The results were evaluated and referred to five age subgroups and duration of hospitalization. The presence of potentially enteropathogenic bacteria was noted in 87 per cent of children in widely varying quantities. In the most numerous group of children the same bacteria were revealed at the beginning and at the end of hospitalization. The microorganisms were most frequently present at the moment of hospitalisation in newborn children. Complete elimination during stay in hospital occurred mainly in the youngest children, who did not acquire these bacteria during their stay in hospital. Colonisation with potentially pathogenic strains in hospital affected mainly children aged 7 to 12 months. EPEC prevailed among the strains isolated in both analyses.

[The occurrence of viral respiratory tract infections in small children in the season 1993-1994 (with reference to the role of breast feeding)]. / Wystepowanie wirusowych zakazen dróg oddechowych u malych dzieci w sezonie 1993-1994 (z uwzglednieniem wplywu karmienia naturalnego).

In the period October 1993 to June 1994, 231 hospitalized children under 2 years of age were examined by immunofluorescence for respiratory viruses. In 114 (49.9%) cases viral etiology was established. The data were compared with results obtained from the season 1992-93. RS virus was predominant as in the previous season but infections of parainfluenzae type 3 virus were rather rare (p < 0.005). Mixed and adenovirus (p < 0.02%) infections were more frequent than in the past season. A breast-fed children were statistical more rarely infected by respiratory viruses (p < 0.05), (RS virus-p < 0.1), than artificial-fed ones.

[Content of immunoglobulin in women's milk during the year after childbirth]. / Zawartosc immunoglobulin w pokarmie kobiet w ciagu roku po porodzie.

Immunoglobulin G during a normal pregnancy passes through the placenta to the fetal blood. The study aimed to evaluate the amount of immunoglobulins which may be transferred with maternal milk during one year following labour. In 1982-1986, 210 mothers who delivered healthy babies have been studied. A content of immunoglobulins was assayed in the colostrum and breast milk (after 1, 3, 6 and 12 months following labour). With radial diffusion technique described by Mancini et al. The results were expressed in mg% and analysed statistically. There were statistically significant differences in the mean IgA, IgG, and IgM levels in the maternal milk in various periods of lactation. No marked effect of passed maternal diseases during pregnancy (including infections) on the mean levels of the above immunoglobulins was noted one month after labour.

Respiratory syncytial virus (RSV) antibodies in different immunoglobulin classes in small children.

Percent of RSV antibodies in the individual immunoglobulin classes and IgG subclasses was determined by ELISA and the immune response to RSV infection was evaluated in children 0-2 years of age. In the first 6 months percent of sera containing IgG-RSV declined to reach minimal in the 7th-9th month, then it rose to 100% in children 2 years of age. Percent of positive IgM-RSV and IgA-RSV gradually increased with age. The IgE antibodies occurred in highest percent (20%) in the youngest children, and showed a tendency to decrease in the next age groups. The IgG-1 antibodies were found in the highest percent of sera, the incidence of other subclass antibodies being lower. Diagnostic increase in antibodies was found in highest number in the IgG class and IgG-1 subclass. In the IgM class the highest percent of antibody increase was detected in 21%, and in the IgE class in about 9%.

The immune response of small children by antibodies of different classes to respiratory syncytial virus (RSV) proteins.

A group of 18 paired sera, obtained from RSV-infected children were assayed by Western-Blot for antibodies for RSV proteins in the immunoglobulin classes IgG, IgM, IgA and IgE and in the IgG subclasses. The IgG antibodies for the protein G were found in almost half of the sera tested, while in the other immunoglobulin classes the antibodies occurred in a much lower number of the sera. The antibodies for protein F1 were detected first of all in the IgG class and subclasses. The antibodies for protein N were present in the IgG class of about half of sera and antibodies for protein P were found first of all in the IgG and IgM classes. A relatively high number of the sera contained IgE antibodies for protein M. In the first 3 months of life, the children poorly responded to the infection by the production of antibodies for RSV proteins. The most clear response by various immunoglobulin classes to RSV proteins was observed in children 4-12 months of age.

[Etiologic profile of viral respiratory infections in small children during epidemic periods 1990-91 and 1991-92]. / Profil etiologiczny wirusowych zakazen dróg oddechowych u malych dzieci w sezonach epidemicznych 1990-91 i 1991-92.

In periods from October 1990 to June 1991 and from September 1991 to June 1992, 551 hospitalized infants and small children were examined by immunofluorescence method for respiratory viruses. In 264 (47.9%) cases viral etiology was established. Like in previous seasons the infections of RS and parainfluenza type 3 viruses dominated. Infections of RS virus were not frequent, and of parainfluenza type 3 virus less than in previous seasons. The increase of percent of infections due to parainfluenza type 2 and 4 viruses were observed.

Plasmid vectors for selecting IS1-promoted deletions in cloned DNA: sequence analysis of the omega interposon.

We have constructed two plasmid vectors which allow selection for in vivo deletions within cloned DNA fragments. The plasmids are derivatives of pBR322 which carry the Escherichia coli rpsL (strA) gene, known to confer a dominant streptomycin (Sm)-sensitivity phenotype to the host cell, and a copy of the IS1 transposable element. Sm-resistant strains that harbor these plasmids display sensitivity to Sm. Spontaneous IS1-promoted deletions across the rpsL gene can be isolated simply by selection for Sm resistance. Hence, nested sets of deletions of a cloned DNA can be obtained and sequenced with an IS1-specific primer. Using this approach, we have determined the complete nucleotide sequence of the omega interposon [Prentki and Krisch, Gene 29 (1984) 303-313].