[Serotype distribution of Streptococcus pneumoniae isolated from invasive infections at the Hospital de Niños of Santa Fe]. / Distribución de serotipos de Streptococcus pneumoniae aislados de infecciones invasoras en el Hospital de Niños de Santa Fe.

The serotype distribution of Streptococcus pneumoniae varies through time. The introduction of pneumococcal conjugate vaccines showed a decreased prevalence of pneumococcal invasive isolates belonging to serotype 14 and an increase of serotypes not therein included. In 1993, the Hospital de Niños of Santa Fe began surveillance of the serotype distribution of invasive S. pneumoniae disease. In the period 2003-2005, 76 isolates were analysed by studying the correlation between serotype and pathology, age and MIC of penicillin. Serotype 14 was the most frequent followed by serotypes 1, 6B, 18C, 7F, 19 F and 5. Serotype 14 showed a statistically significant correlation with MICs of penicillin ranging from 0,5 to 2 mg/l. Although this serotype was more frequently observed in pneumonia than in meningitis, there was not a significant association with any particular pathology. Serotypes 14 and 1, were prevalent among children under and over 2 years old, respectively. Most of these isolates with MICs of penicillin = 2 mg/l, were from patients with pneumonia and not with meningitis. The serotype distribution was similar to that during the period 1993-99, with the exception of serotypes 18C, 4, 12F and 22F which had never been found before. The emergence of these serotypes makes it essential to continue surveillance to determine which conjugated vaccine formulation would be suitable to prevent the most frequent pneumococcal invasive infections.

Identification of galectin-3 and mucin-type O-glycans in breast cancer and its metastasis to brain.

Galectin-3 has been implicated in tumor progression. We demonstrated immunohistochemically that galectin-3 was negative in normal breast tissue, but it was highly increased in breast cancer and in metastatic tissues to brain. Similarly, histochemistry with mucin-specific lectins showed increased recognition in breast tumor and metastasis with Machaerocereus eruca agglutinin (Fualpha 1,2 (GalNAcalpha 1,3) Galss1,4 in complex mucin) but not for Amaranthus leucocarpus (Galss1,3-GalNAc-alpha 1,0-Ser/Thr) and Arachis hypogaea lectins (Galss1,3GalNAc/Galss1,4GlcNAc). Mucin-type glycans and galectin-3 colocalized in breast cancer and metastasis, but not in normal tissue, suggesting upregulated biosynthesis of complex O-glycosidically linked glycans and galectin-3 favor breast cancer progression and brain metastasis.

Distribución de serotipos de Streptococcus pneumoniae aislados de infecciones invasoras en el Hospital de Niños de Santa Fe / Serotype distribution of Streptococcus pneumoniae isolated from invasive infections at the Hospital de Niños of Santa Fe.

Con la introducción de vacunas conjugadas antineumocócicas se observó, en muchos países, disminución de aislamientos de Streptococcus pneumoniae del serotipo 14 y aumento de aislamientos correspondientes a serotipos no incluidos en esas vacunas. En 1993, el Hospital de Niños de Santa Fe comenzó la vigilancia de la distribución de serotipos de Streptococcus pneumoniae invasores. En este trabajo se estudió la correlación entre serotipo y a) patología (neumonía/meningitis), b) edad (menor o mayor de dos años), y c) CIM de penicilina, para los serotipos aislados en el período 2003-2005. El serotipo predominante fue el 14, seguido del 1, 6B, 18C, 7F, 19F y 5. El serotipo 14 mostró asociación estadísticamente significativa con valores de CIM de penicilina entre 0,5 y 2 mg/l, no así con alguna patología, aunque se lo halló con mayor frecuencia en neumonías que en meningitis. Los serotipos 14 y 1 prevalecieron en niños menores y mayores de 2 años, respectivamente. La CIM de penicilina = 2 mg/l se observó más en neumonías que en meningitis. La frecuencia relativa de los diferentes serotipos hallados fue semejante a la observada en el período 1993-99; no obstante, los serotipos 18C, 4, 12F y 22F no se habían encontrado antes. La aparición de nuevos serotipos convierte en importante la vigilancia, dada la necesidad de formular vacunas que los incluyan y que efectivamente prevengan las infecciones neumocócicas más comunes.

The serotype distribution of Streptococcus pneumoniae varies through time. The introduction of pneumococcal conjugate vaccines showed a decreased prevalence of pneumococcal invasive isolates belonging to serotype 14 and an increase of serotypes not therein included. In 1993, the Hospital de Niños of Santa Fe began surveillance of the serotype distribution of invasive S. pneumoniae disease. In the period 2003 - 2005, 76 isolates were analysed by studying the correlation between serotype and pathology, age and MIC of penicillin. Serotype 14 was the most frequent followed by serotypes 1, 6B, 18C, 7F, 19 F and 5. Serotype 14 showed a statistically significant correlation with MICs of penicillin ranging from 0,5 to 2 mg/l. Although this serotype was more frequently observed in pneumonia than in meningitis, there was not a significant association with any particular pathology. Serotypes 14 and 1, were prevalent among children under and over 2 years old, respectively. Most of these isolates with MICs of penicillin = 2 mg/l, were from patients with pneumonia and not with meningitis. The serotype distribution was similar to that during the period 1993-99, with the exception of serotypes 18C, 4, 12F and 22F which had never been found before. The emergence of these serotypes makes it essential to continue surveillance to determine which conjugated vaccine formulation would be suitable to prevent the most frequent pneumococcal invasive infections.

[Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis]. / Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística.

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7%), P. aeruginosa (37.4%) and Haemophilus spp (15.3%). A high resistance to erithromycine (35.0%) and clindamicine (29.4%) was observed in S. aureus strains and 25.9% of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0%). The rate of ampicillin-resistant Haemophilus spp. was 23.0% and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0%).

Evaluation of a nested-PCR assay for Streptococcus pneumoniae detection in pediatric patients with community-acquired pneumonia / Evaluación de una PCR anidada en pacientes pediátricos para diagnóstico de neumonía neumocócica adquirida en la comunidad

The aim of the present work was to evaluate the usefulness of a simplified method for DNA extraction coupled to a nested-PCR protocol, based on the amplification of pneumolysin gene fragments for the diagnosis of pneumococcal pneumonia in pediatric patients with clinical and radiological evidence of bacterial infection. Bacterial DNA was extracted from sera by boiling and used without further purification in the PCR for the pneumolysin gene. None toxic reagents were used and the necessary steps to obtain the DNA were left at a minimum; furthermore, it overcomes the use of expensive commercial kits for DNA purification. The total procedure can be completed the same day of sampling and, most important, it avoids the use of sophisticated technology. Both in vitro analytical specificity and sensitivity (10 CFU/ml) of the assay were similar to those previously reported. When clinical samples were tested, the rate of positivity was shown to be 83.3% and 71% in pediatric patients with positive (group a) and negative blood cultures (group b), respectively. In group a, DNA detection was successful in samples from children without treatment or with less than 48 h of antibiotic therapy. None amplification was obtained from sera patients with viral infection or in samples from healthy controls. The application of the strategy described in this paper substantially seems to improve the diagnostic process in a determinate group: blood culture-negative children with pneumonia.

El objetivo del presente trabajo fue evaluar la utilidad de un método simplificado para extracción de ADN, acoplado a un protocolo de PCR anidada, basada en la amplificación de fragmentos del gen de la neumolisina para el diagnóstico de neumonía neumocócica en niños con evidencias clínicas y radiológicas de infección bacteriana. El ADN bacteriano fue extraído del suero por calentamiento y utilizado en la PCR para el gen de la neumolisina sin purificación posterior. Para la obtención de ADN no se utilizan reactivos tóxicos ni costosos "kits" comerciales. El procedimiento completo puede ser realizado en el día y lo que es más importante, evita el uso de tecnología sofisticada. La especificidad analítica in vitro y la sensibilidad (10 UFC/ml) del ensayo fueron similares a lo hallado en publicaciones anteriores. El porcentaje de muestras positivas fue del 83,3% y del 71% en los pacientes con hemocultivos positivos (grupo a) y negativos (grupo b), respectivamente. En el grupo a, sólo se obtuvieron resultados positivos mediante la PCR anidada en los pacientes no tratados o con menos de 48 hs de tratamiento antibiótico. No se obtuvieron señales de amplificación en los sueros de los pacientes con infecciones virales ni en las muestras del grupo control. La aplicación de la estrategia descripta incrementa la posibilidad diagnóstica de neumonía neumocócica en niños con hemocultivos negativos.

Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística / Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis

La fibrosis quística (FQ) se caracteriza por disfunciones en las glándulas de secreción exocrina del organismo. Las primeras manifestaciones suelen observarse en el sistema respiratorio, constituyendo una de las causas más importantes de morbimortalidad en los pacientes afectados. Los microorganismos patógenos que colonizan frecuentemente el tracto respiratorio de estos pacientes son Staphylococcus aureus, Haemophilus spp., y Pseudomonas aeruginosa. Entre noviembre de 2001 y agosto de 2004 se estudiaron 222 muestras respiratorias de pacientes con FQ de entre 4 meses y 11 años de edad. Se aislaron S. aureus (38,7%), P. aeruginosa (37,4%) y Haemophilus spp., (15,3%). En S. aureus la meticilina-resistencia fue del 25,9% y se asoció con altas resistencias a eritromicina (35,0%) y clindamicina (29,4%). El mayor porcentaje de resistencia observado en las cepas de P. aeruginosa fue frente a gentamicina (31,0%). Los aislamientos de Haemophilus spp. fueron resistentes a ampicilina (23,0%) debido a la presencia de beta-lactamasas, y a trimetoprima/sulfametoxazol (59,0%).

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7%), P. aeruginosa (37.4%) and Haemophilus spp (15.3%). A high resistance to erithromycine (35.0%) and clindamicine (29.4%) was observed in S. aureus strains and 25.9% of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0%). The rate of ampicillin-resistant Haemophilus spp. was 23.0% and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0%).

Evaluation of a nested-PCR assay for Streptococcus pneumoniae detection in pediatric patients with community-acquired pneumonia.

The aim of the present work was to evaluate the usefulness of a simplified method for DNA extraction coupled to a nested-PCR protocol, based on the amplification of pneumolysin gene fragments for the diagnosis of pneumococcal pneumonia in pediatric patients with clinical and radiological evidence of bacterial infection. Bacterial DNA was extracted from sera by boiling and used without further purification in the PCR for the pneumolysin gene. None toxic reagents were used and the necessary steps to obtain the DNA were left at a minimum; furthermore, it overcomes the use of expensive commercial kits for DNA purification. The total procedure can be completed the same day of sampling and, most important, it avoids the use of sophisticated technology. Both in vitro analytical specificity and sensitivity (10 CFU/ml) of the assay were similar to those previously reported. When clinical samples were tested, the rate of positivity was shown to be 83.3% and 71% in pediatric patients with positive (group a) and negative blood cultures (group b), respectively. In group a, DNA detection was successful in samples from children without treatment or with less than 48 h of antibiotic therapy. None amplification was obtained from sera patients with viral infection or in samples from healthy controls. The application of the strategy described in this paper substantially seems to improve the diagnostic process in a determinate group: blood culture-negative children with pneumonia.

Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística. / [Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis]

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7

), P. aeruginosa (37.4

) and Haemophilus spp (15.3

). A high resistance to erithromycine (35.0

) and clindamicine (29.4

) was observed in S. aureus strains and 25.9

of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0

). The rate of ampicillin-resistant Haemophilus spp. was 23.0

and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0

).

Evaluation of a nested-PCR assay for Streptococcus pneumoniae detection in pediatric patients with community-acquired pneumonia.

The aim of the present work was to evaluate the usefulness of a simplified method for DNA extraction coupled to a nested-PCR protocol, based on the amplification of pneumolysin gene fragments for the diagnosis of pneumococcal pneumonia in pediatric patients with clinical and radiological evidence of bacterial infection. Bacterial DNA was extracted from sera by boiling and used without further purification in the PCR for the pneumolysin gene. None toxic reagents were used and the necessary steps to obtain the DNA were left at a minimum; furthermore, it overcomes the use of expensive commercial kits for DNA purification. The total procedure can be completed the same day of sampling and, most important, it avoids the use of sophisticated technology. Both in vitro analytical specificity and sensitivity (10 CFU/ml) of the assay were similar to those previously reported. When clinical samples were tested, the rate of positivity was shown to be 83.3

and 71

in pediatric patients with positive (group a) and negative blood cultures (group b), respectively. In group a, DNA detection was successful in samples from children without treatment or with less than 48 h of antibiotic therapy. None amplification was obtained from sera patients with viral infection or in samples from healthy controls. The application of the strategy described in this paper substantially seems to improve the diagnostic process in a determinate group: blood culture-negative children with pneumonia.

[Necrotizing lymphadenitis caused by Nocardia asteroides in a healthy girl]. / Linfoadenitis necrotizante por Nocardia asteroides en una niña previamente sana.

A case of a healthy girl with a trauma in her right leg is described. Twelve hours later she developed fever and pain in the inguinal region. Two days later, she was admitted to the Hospital, and there, an aspiration of pus from the lymph node was performed, and this sample was sent for culture. The Gram smear showed gram-positive filamentous bacilli and when Kinyoun was used they were observed as weakly acid fast rods. After four days of incubation under aerobic conditions, white and hemolitic colonies were seen on human blood agar plates. Biochemical tests as urea hydrolysis and gelatin liquefaction were performed. To complete the identification it was sent to a reference laboratory, INEI-ANLIS Dr. Carlos G. Malbrán, where the isolate was confirmed as Nocardia asteroides sensu stricto. The patient was treated with intravenous cephalotin with good evolution. Therapy was followed with oral trimethoprim-sulfamethoxazole. This case is the first report of a Nocardia infection in the Hospital de Niños de Santa Fe. The fact that N. asteroides was isolated from an immunologically competent girl should be highlighted. This species is uncommon in children and it is rare in our region. In addition, N. asteroides infections are usually seen in pulmonary disease and rarely producing cutaneous infections.

Linfoadenitis necrotizante por Nocardia asteroides en una niña previamente sana / [Necrotizing lymphadenitis caused by Nocardia asteroides in a healthy girl]

A case of a healthy girl with a trauma in her right leg is described. Twelve hours later she developed fever and pain in the inguinal region. Two days later, she was admitted to the Hospital, and there, an aspiration of pus from the lymph node was performed, and this sample was sent for culture. The Gram smear showed gram-positive filamentous bacilli and when Kinyoun was used they were observed as weakly acid fast rods. After four days of incubation under aerobic conditions, white and hemolitic colonies were seen on human blood agar plates. Biochemical tests as urea hydrolysis and gelatin liquefaction were performed. To complete the identification it was sent to a reference laboratory, INEI-ANLIS Dr. Carlos G. Malbrán, where the isolate was confirmed as Nocardia asteroides sensu stricto. The patient was treated with intravenous cephalotin with good evolution. Therapy was followed with oral trimethoprim-sulfamethoxazole. This case is the first report of a Nocardia infection in the Hospital de Niños de Santa Fe. The fact that N. asteroides was isolated from an immunologically competent girl should be highlighted. This species is uncommon in children and it is rare in our region. In addition, N. asteroides infections are usually seen in pulmonary disease and rarely producing cutaneous infections.

Linfoadenitis necrotizante por Nocardia asteroides en una niña previamente sana. / [Necrotizing lymphadenitis caused by Nocardia asteroides in a healthy girl]

A case of a healthy girl with a trauma in her right leg is described. Twelve hours later she developed fever and pain in the inguinal region. Two days later, she was admitted to the Hospital, and there, an aspiration of pus from the lymph node was performed, and this sample was sent for culture. The Gram smear showed gram-positive filamentous bacilli and when Kinyoun was used they were observed as weakly acid fast rods. After four days of incubation under aerobic conditions, white and hemolitic colonies were seen on human blood agar plates. Biochemical tests as urea hydrolysis and gelatin liquefaction were performed. To complete the identification it was sent to a reference laboratory, INEI-ANLIS Dr. Carlos G. Malbrán, where the isolate was confirmed as Nocardia asteroides sensu stricto. The patient was treated with intravenous cephalotin with good evolution. Therapy was followed with oral trimethoprim-sulfamethoxazole. This case is the first report of a Nocardia infection in the Hospital de Niños de Santa Fe. The fact that N. asteroides was isolated from an immunologically competent girl should be highlighted. This species is uncommon in children and it is rare in our region. In addition, N. asteroides infections are usually seen in pulmonary disease and rarely producing cutaneous infections.

Alternating antipyretics: is this an alternative?

OBJECTIVES: To identify current fever management strategies and their basis, and to assess the frequency of alternating acetaminophen and ibuprofen. BACKGROUND: Practicing pediatricians confront the dilemma of elevated temperature and the anxiety this creates for parents. An informal survey of pediatricians revealed a variety of management strategies, including alternating acetaminophen and ibuprofen. There are no scientific data regarding alternating these 2 products. Design. During professional meetings, pediatricians voluntarily filled out a 15-item questionnaire. RESULTS: One hundred sixty-one completed surveys were reviewed. Respondents were mostly general pediatricians (67.7%), with >/=20 years in practice (55.9%). Most respondents chose a temperature of 101(o)F to start antipyretic treatment (61.9%). A small percentage used discomfort alone as the indication (13%). The antipyretic of choice was temperature-dependent in 50% of respondents; 57% used ibuprofen for temperature >/=102 degrees F. Fifty percent of respondents advised parents to alternate acetaminophen and ibuprofen. The method of alternation varied. The most common answers given for choosing a particular antipyretic regime were recommendations of the American Academy of Pediatrics (29%) and opinions of colleagues and mentors (25%). Physicians with <5 years of practice were significantly more likely to alternate antipyretics (69.7%). CONCLUSION: Acetaminophen and ibuprofen are commonly being used in an alternating manner for management of fever. There is presently no scientific evidence that this combination is safe or achieves faster antipyresis than either agent alone. There is evidence that the improper use of these agents may cause harm. Despite 29% of participants citing American Academy of Pediatrics recommendations as the basis for fever management, no such policy or recommendations exist. The observation that this practice is more common in younger practitioners may reflect their continued anxiety about fever (fever phobia). Until properly controlled studies have assessed the risk of combining these 2 products, practitioners should proceed with caution.

Genetic, clinical, and biochemical analysis of unrelated Spanish families with multiple endocrine neoplasia type I.

OBJECTIVE: (1) To study seven unrelated Spanish families with multiple endocrine neoplasia type I (MEN I), describing clinical features and investigating the presence of germline mutations in the MEN1 gene, and (2) to establish reference values for pancreatic polypeptide and gastrin after a standardized test meal in a healthy control group, analyzing the usefulness of this test for detecting neuroendocrine gastroenteropancreatic tumors in subjects with MEN I. METHODS: Two or three generations of 7 kindreds with MEN I, consisting of a total of 39 individual family members, were investigated. Three of the families were subjected only to genetic analysis, and the other four families were also assessed clinically. A group of 23 healthy control subjects were also studied. RESULTS: Mutations in the MEN1 gene were found in six of the seven families studied. Of the 4 families studied clinically, 12 family members were genetically affected. In these study subjects, hyperparathyroidism, adrenal adenomas, neuroendocrine gastroenteropancreatic tumors, and pituitary adenomas developed in 100%, 50%, 16%, and 12%, respectively. All demonstrated pancreatic tumors were associated with abnormal results after a test meal, but 75% of them also showed high basal hormonal measurements. CONCLUSION: Analysis of the MEN1 gene decreases the total number of subjects who need to undergo repeated clinical and biochemical studies, but genetic mutations are not detected in all families with MEN I. Hyperparathyroidism is the most common manifestation of the syndrome, but the presence of adrenal adenomas has probably been underestimated. Ingestion of a standardized test meal for stimulation of gastrin and pancreatic polypeptide could be a complementary procedure for diagnosing gastroenteropancreatic tumors in selected patients with MEN I in whom basal gastrin and pancreatic polypeptide levels are normal.

Molecular pathology of multiple endocrine neoplasia type I: two novel germline mutations and updated classification of mutations affecting MEN1 gene.

Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterized by the combined development of tumors in several endocrine glands and other tissues. The MEN1 gene was recently identified and isolated by positional cloning. This gene was screened in two unrelated MEN1 Spanish kindreds (with four affected members and seven asymptomatic members) using single-strand conformation polymorphism, DNA sequencing, and restriction enzyme analysis. Two novel germline mutations were identified: a missense in exon 2 (H139R) and a splice-site in intron 9 (1461-2A>C). These findings allowed us to identify the MEN1 carriers among the seven asymptomatic members analyzed. An updated review of the mutations and polymorphisms found in the analysis of the MEN1 gene is provided. The report of all germline mutations causing MEN1 and easy access to this updated information are both of special diagnostic interest, because this greatly facilitates the task of attributing the disorder to a specific mutation found in a given MEN1 family. This is especially helpful in the critical differentiation of missense mutations from nonsynonymous polymorphisms that fit the pattern of segregation of the disease, but do not cause it.

A novel germline mutation in exon 5 of the multiple endocrine neoplasia type 1 gene.

The autosomal dominant multiple endocrine neoplasia type 1 (MEN1) syndrome is characterized by neoplasia of parathyroids, anterior pituitary, and gastrointestinal and pancreatic neuroendocrine tissues. Recently the gene responsible for the MEN1 syndrome has been identified on chromosome region 11q13. Most of the described mutations are nucleotide substitutions and small deletions affecting exons 2 and 3, causing protein truncation. Only one mutation in exon 5 has been found, and this corresponds to a MEN1 sporadic case. Small insertions are also rare. We studied a MENI family composed of five members, two of whom were clinically affected. We found a new germline 1 basepair insertional mutation affecting the exon 5 of the MEN1 gene in the two members affected in this MEN1 family.

Salmonella Freetown: primer aislamiento en Argentina / Salmonella Freetown: first report in Argentina

En el presente trabajo se comunica el primer aislamiento de Salmonella Freetown en Argentina. Se obtuvo a partir de un coprocultivo de una paciente de 1 mes y 7 días, cuya madre consultó por un cuadro de diarrea aguda en el Hospital de Niños Dr. Ricardo Gutiérrez de la ciudad de Santa Fe. Por sus características bioquímicas y su fórmula antigénica, esta nueva serovariedad se ubica dentro de la especie Salmonella enterica subespecie enterica