Detection of human immunodeficiency virus type 1 (HIV-1) in heel prick blood on filter paper from children born to HIV-1-seropositive mothers.

The human immunodeficiency virus type 1 (HIV-1) DNA PCR results of 94 dried blood spot (DBS) samples on filter paper and corresponding venous blood in EDTA obtained from infants born to HIV-1-seropositive mothers were compared. In addition, the results of HIV-1 DNA PCR on DBS and the HIV-1 RNA PCR from plasma of 70 paired samples were compared. A 100% specificity and a 95% sensitivity for HIV-1 DNA PCR on DBS compared with results for venous blood were observed for the 94 paired samples. The results of the DBS HIV-1 DNA PCR and HIV-1 RNA PCR of 70 corresponding plasma samples correlated perfectly (100%). The DBS HIV-1 DNA PCR method proved reliable for HIV-1 detection.     

Detection and subtyping of HIV-1 isolates with a panel of characterized monoclonal antibodies to HIV p24gag

A panel of monoclonal antibodies (Mabs) was used to analyze the number and localization of B-cell epitopes on human immunodeficiency virus (HIV) p24gag and the variability of these epitopes in sequential HIV isolates and in isolates from different geographical origin. The specificity of these Mabs was demonstrated by immunoblotting and radioimmunoprecipitation assays. Cross-inhibition experiments indicated the presence of at least five different epitopes on p24. Analysis with p24 recombinant products revealed that three of the Mabs to p24 were directed to epitopes localized on the C-terminal part. Four other Mabs were directed to epitopes localized on the N-terminal half of the protein. Anti-p24 Mabs were used to develop HIV p24 antigen-capture assays. Application of these assays in HIV isolation resulted in more efficient recovery of HIV. Serotyping of HIV-1 isolates with five anti-p24 Mabs demonstrated that 55/65 isolates recovered from Dutch and Belgian individuals, but only 4/9 HIV-1 African isolates, were recognized by all five Mabs. Five of nine Central African HIV-1 isolates were not reactive with at least one of these Mabs. The variability of p24 appeared to be predominantly localized on the N-terminal part of the protein. Lack of expression of antigenic determinants on p24 was shown to be independent of culture conditions. Moreover, an infectious molecular clone was shown to have the same serotype as the corresponding HIV isolate. The serotype of sequential isolates obtained from 17 individuals over a 1 1/2- to 2 1/2-year period did not change, suggesting a limited in vivo p24 variation over time.     

Chip-based mtDNA mutation screening enables fast and reliable genetic diagnosis of OXPHOS patients.

PURPOSE: Oxidative phosphorylation is under dual genetic control of the nuclear and the mitochondrial DNA (mtDNA). Oxidative phosphorylation disorders are clinically and genetically heterogeneous, which makes it difficult to determine the genetic defect, and symptom-based protocols which link clinical symptoms directly to a specific gene or mtDNA mutation are falling short. Moreover, approximately 25% of the pediatric patients with oxidative phosphorylation disorders is estimated to have mutations in the mtDNA and a standard screening approach for common mutations and deletions will only explain part of these cases. Therefore, we tested a new CHIP-based screening method for the mtDNA. METHODS: MitoChip (Affymetrix) resequencing was performed on three test samples and on 28 patient samples. RESULTS: Call rates were 94% on average and heteroplasmy detection levels varied from 5-50%. A genetic diagnosis can be made in almost one-quarter of the patients at a potential output of 8 complete mtDNA sequences every 4 days. Moreover, a number of potentially pathogenic unclassified variants (UV) were detected. CONCLUSIONS: The availability of long-range PCR protocols and the predominance of single nucleotide substitutions in the mtDNA make the resequencing CHIP a very fast and reliable method to screen the complete mtDNA for mutations.     

Heterogeneous malignant non Hodgkin's lymphomas as a causative disorder in lethal midline granuloma

Summary The present report describes the results of a combined morphological, enzyme- and immunohistochemical analysis of nine cases of malignant non Hodgkin's lymphomas (NHL) clinically presenting as lethal midline granuloma. In a previous report written before antibodies directed against B and T lymphocytes were available, a histiocytic origin of such neoplasms had been suggested. A panel of antibodies reactive with most B cells (L26, MB1, KiB3) and a majority of T cells (MT1, UCHL1) was applied on paraffin sections of formalin fixed tissues as well as antibodies directed against leukocyte common antigen (LCA), myeloid/histiocyte antigen (MAC 387), lysozyme, alpha-1-antitrypsin, alpha-1-antichymotrypsin, S-100 protein, prekeratin and immunoglobulin light chains. Enzyme histochemistry included tests for non-specific acid esterase, acid phosphatase, betaglucuronidase and chloroacetate esterase. As a result, five T, two B and two unclassified (malignant histiocytosis probable) NHL were identified, indicating distinct heterogeneity of NHL as causative disorders in lethal midline granuloma.     

Expression of calcium-binding proteins MRP8 and MRP14 in inflammatory muscle diseases

The pathophysiological role of infiltrating macrophages and their subtypes in idiopathic inflammatory myopathies such as dermatomyositis, polymyositis, and inclusion body myositis is not fully clear. Monocytes exhibit various phenotypes with different functional properties such as release of pro- or anti-inflammatory mediators. Expression of myeloid-related proteins MRP8 and MRP14, two calcium-binding S100-proteins, characterizes a proinflammatory subtype of macrophages. We immunohistochemically investigated expression of MRP8 and MRP14 in muscle biopsies of 33 patients with dermatomyositis, polymyositis, and inclusion body myositis. We found a clear association of expression of MRP8 and MRP14 by infiltrating macrophages with degeneration of myofibers. Because MRP8 and MRP14 are secreted by activated macrophages we investigated if these proteins would have direct extracellular effects on myocytes. We found that the purified MRP8/MRP14 complex inhibited proliferation and differentiation of C2C12 myoblasts and that it induced apoptosis via activation of caspase-3 in a time- and dose-dependent manner. These results indicate that in the course of inflammatory myopathies, activated macrophages can promote destruction and impair regeneration of myocytes via secretion of MRP8/MRP14.     

Liver-First Approach for Synchronous Colorectal Metastases: Analysis of 7360 Patients from the LiverMetSurvey Registry

Annals of Surgical Oncology - The liver-first approach in patients with synchronous colorectal liver metastases (CRLM) has gained wide consensus but its role is still to be clarified. We aimed to...     

Effects of intravenously administered hypertonic urea solution

Summary and Conclusions In 12 patients with increased intracranial pressure, caused by an expanding process, a hypertonic urea solution was intravenously administered during a craniotomy. At different times before, during and after the operation, the electrolytes, urea, glucose and total protein values were determined in various body fluids and tissues.This study disclosed that the urea administered was distributed through both the intracellular and the extracellular space after 20 minutes. The values of the electrolytes, except the calcium, in the extracellular fluid remained constant after administration of the urea solution; the total protein value, however, showed a considerable decrease.It was established that the blood-brain barrier plays no appreciable role in the mechanism of action of hypertonic urea solutions in dehydrating the brain tissue; the blood-C. S. P. and brain-C. S. F. barriers may do.

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Zusammenfassung Bei 12 Patienten mit intrakranieller Drucksteigerung infolge eines raumbeengenden Prozesses wurde Harnstofflösung intravenös während der Schädeleröffnung gegeben. Zu verschiedenen Zeitpunkten vor, während und nach der Operation wurden Elektrolyt, Harnstoff, Glukose und Gesamteiweiß quantitativ bestimmt und zwar sowohl in verschiedenen Körperflüssigkeiten wie auch in Geweben.Die Untersuchungen ergaben, daß der verabfolgte Harnstoff in 20 Minuten sich sowohl auf den intrazellulären, wie den extrazellulären Raum verteilt hat. Die Elektrolytwerte, mit Ausnahme von Kalzium, blieben nach der Harnstoffinfusion in den extrazellulären Flüssigkeiten unverändert, der Gesamteiweißwert nahm dagegen beträchtlich ab.Es wurde festgestellt, daß die Bluthirnschranke keine wesentliche Rolle für die entwässernde Wirkung des Harnstoffes auf das Hirngewebe spielt, während die Blut-Liquor-Schranke und die Hirn-Liquor-Schranke vielleicht von Bedeutung sind.

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Resumen Después de una craniectomía se administró una solución de urea hipertónica por via intravenosa a 12 pacientes que presentaban una presión intracraneal creciente a causa de una exposición de la hipófisis. Periódicamente, antes, durante y después de la operación se determinaron los valores de los electrolitos, de la urea, de la glucosa y de las proteinas totales en los diferentes líquidos y tejidos del organismo.Este estudio demostróque la urea administrada se distribuia a través del espacio intra y extracelular al cabo de 20 minutos. Los valores de los electrolitos, excepto el calcio, permanecieron constantes en el líquido extracelular después de la administración de la solución de urea; el valor de las proteinas totales, sin embargo, mostró un descenso considerable.Se concluyó que la barrera hemato-encefálica no juega ningún papel apreciable en los mecanismos de acción de las soluciones de urea hipertónica en la deshidratación del tejido cerebral; tal vez lo juegue en las barreras sangre-liquido cofalo-raquídeo y cerebro-líquido cefalo-raquídeo.

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Résumé Lors d'une craniotomie, une solution d'urée hypertonique fut administrée par voie intraveineuse chez 12 patients présentant une pression intracrânienne grandissante causée par une expansion de l'apophyse. De temps en temps, avant, pendant et après l'opération, les valeurs des électrolytes, de l'urée, du glucose et de la protéine totale étaient déterminées dans les différents liquides et tissus du corps.Cette étude démontra que l'urée administrée était distribuée à travers l'espace intra et extraecllulaire au bout de 20 minutes. Les valeurs des électrolytes, excepté le calcium, demeurèrent constantes dans le liquide extracellulaire après l'administration de la solution d'urée; la valeur de la protéïne totale, pourtant, montrait une baisse considérable.Il fut établi que la barrière hémato-encéphalique ne joue aucun rôle appréciable dans le mécanisme d'action des solutions d'urée hypertonique dans la déshydratation du tissu cérébral; les barrières sang-liquide céphalorachidien et cerveau-liquide céphalo-rachidien, peut-être.

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Riassunto In 12 pazienti con ipertensione endocranica, causata da un processo espansivo, è stata somministrata durante la craniotomia dell'urea in soluzione ipertonica per via venosa. A diversi tempi prima, durante e dopo l'intervento, sono stati dosati gli elettroliti, l'urea, il glueosio e le proteine totali in vari fluidi e tessuti corporei. Queste ricerche hanno evidenziato che l'urea viene distribuita tra spazio intracellulare ed extracellulare in 20 minuti. I livelli degli elettroliti, eccetto il calcio, rimangono costanti nel liquido extracellulare dopo la somministrazione di urea, i valori della proteinemia totali invece mostrano una notevole diminuizione.E' stato stabilito che la barriera emato-cerebrale non gioca alcun ruolo apprezzabile nel meccanismo d'azione dell'urea ipertonica nel disidratare il tessuto cerebrale; un ruolo importante potrebbe essere inveoe giocato dalla barriera emato-liquorale e tra liquor e sistema nervoso.

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This study was supported by a grant from the Netherlands Organization for the Advancement of Pure Research (Nederlandse Organisatie voor Zuiver-Wetenschappelijk Onderzoek, Z. W. O.).