Cytogenetic analysis of 100 consecutive newly diagnosed cases of acute lymphoblastic leukemia in Rio de Janeiro

We report the cytogenetic analysis of newly diagnosed Brazilian children with acute lymphocytic leukemia (ALL). We investigated 100 ALL cases from four different institutions in Rio de Janeiro. The frequency of chromosomal abnormalities was 92.3%. The karyotype profile and recurrent abnormalities found in this study do not differ essentially from those described by other groups. Although the Brazilian population is usually the product of different ethnic groups, our results show that the frequency of each recurrent abnormality is similar to that found in populations without our degree of diverse ethnic composition. Hence, our results suggest that childhood ALL in Brazil has the same biological features as that in developed countries, supporting the use of similar treatment protocols. We can therefore expect to reach the same survival rates in the coming years, depending possibly on the efficacy of the support therapy and extent of social assistance.     

Control of pulmonary vascular smooth muscle tone by sarcoplasmic reticulum ca2+ pump blockers: thapsigargin and cyclopiazonic acid

The effect of thapsigargin (TG) and cyclopiazonic acid (CPA) on the mechanical activity of the rat pulmonary artery were investigated. In chemically (-escin)-skinned arterial strips, application of TG (0.1–1 M) or CPA (0.5–10 M) prior and throughout the loading procedure of the internal Ca²⁺ stores (0.3 M free Ca²⁺ ions for 8–10 min) concentration dependently inhibited the subsequent contractile response induced by noradrenaline (NA, 10 M) or caffeine (25 mM). In intact strips repeatedly incubated in a Ca²⁺-containing solution (2.5 mM for 10 min), followed by incubation in a Ca²⁺-free solution 12 min before NA-stimulation, TG and CPA not only inhibited the NA-induced contraction but also increased the tension which appeared during the exposure time to Ca²⁺. The two phenomena developed with similar time courses. The increase in tension during the readmission of Ca²⁺ ions was not antagonized by verapamil (10 M) or nifedipine (1 M) but was blocked by La³⁺ (50 M) and Co²⁺ (1 mM) ions. The amplitude of the verapamil-insensitive TG (or CPA)-induced contraction was dependent on the external [Ca²⁺] [0.1–10 mM, concentration for half maximal effect (EC₅₀) =0.85 mM], not modified by the reduction of the external [Na⁺] (from 130 to 10 mM) and decreased by depolarization of the strip using K⁺-rich (30–120 mM) solutions. Under the latter condition, 38±9 and 83±4% reduction (n=5) was observed in the presence of 60 and 120 mM K⁺ respectively. This contraction was also concentration dependently inhibited by the tyrosine kinase inhibitors genistein (0.5–50 M) and tyrphostin (2–50 M). Sr²⁺ ions, which contracted both depolarized intact and skinned strips, failed to replace Ca²⁺ ions in the verapamil-insensitive contraction induced by TG or CPA (n=4). Finally, TG (1 M) and CPA (10 M) did not modify the pCa tension relationship in skinned strips (n=5). These results show that the main action of TG and CPA in rat pulmonary artery is to prevent the refilling of the internal Ca²⁺ store. TG and CPA also seem to facilitate a Ca²⁺ influx through a specific verapamil-insensitive pathway. The biophysical and molecular characteristics of this pathway remain to be elucitated, although it appears to involve a tyrosine kinase activity.     

Laboratory strategies for efficient handling of paraffin-embedded tissues for molecular detection of clonality in non-hodgkin lymphomas.

We herein present a technical strategy to optimize DNA isolation from paraffin-embedded tissue (PET). This includes the choice of adequate buffers for proteinase K digestion and multiplex PCR amplifications for assessing the appropriateness of DNA extracts for subsequent PCR assays for detecting clonality. We found that the association of proteinase K digestion in nonionic buffer and subsequent extract dilutions accounted for 79% of successful amplifications. A final efficiency of 88% was achieved by additional organic extractions and/or re-extractions. Comparisons were carried out with control DNA extracts from fresh samples to assess the efficiency of each clonality assay. Immunoglobulin CDRIII rearranged region amplification was more efficient for pregerminal center B-cell lymphomas in contrast to CDRII rearrangement detection, which was more effective for germinal and postgerminal lymphomas. T-cell clonality detection by TCRgamma PCR was less efficient in PET samples than in fresh tissues showing that DNA integrity is more critical for TCR than for IGH amplification. Two inconclusive cases without phenotypic markers and two other atypical lymphoproliferations masked by reactive T cells were diagnosed as plasmablastic lymphomas and as monoclonal B-proliferations, respectively, due to IGH rearrangements.     

Intracellular cytosolic free calcium concentration in the macula densa and in ascending limb cells at different luminal concentrations of sodium chloride and with added furosemide

The juxtaglomerular apparatus fulfils several important regulatory functions in the kidney, such as tubuloglomerular feedback (TGF) control and control of renin release. The macula densa (MD) cells sense the fluid load by perceiving the distal NaCl concentration via a Na-K-2Cl cotransport system in the luminal cell membrane. It has been proposed that macula densa cell activation may involve changes in intracellular cytosolic free calcium concentration ([Ca2+]i), as one link in the chain of events activating TGF or releasing renin. We therefore investigated the changes in the intracellular calcium concentrations with fura-2, using a video system, in macula densa cells, and compared them with the changes in the corresponding concentrations in the ascending limb of the loop of Henle (c-TAL). The results show that our technique for analysing intracellular cytosolic free calcium in isolated perfused tubules is valid for this purpose, and the Kd value obtained was similar to that found by Grynkiewicz et al. (1985). The intracellular cytosolic free calcium concentration was about 90 nM both in the macula densa and c-TAL cells, and the macula densa cell intracellular cytosolic free calcium concentration increased by about 20 nM when the tubular lumen was perfused with Na and Cl at low concentrations. No significant changes were noted when furosemide was added to the perfusion solutions. We consider it hardly likely that this small change in intracellular cytosolic free calcium concentration can be entirely responsible for full activation of renin release or full inactivation of the TGF control mechanism. It would seem that the signal transmission from the macula densa cells could occur by other routes than through activation of intracellular cytosolic free calcium concentration.     

Transdermal microconduits by microscission for drug delivery and sample acquisition

Background  

Painless, rapid, controlled, minimally invasive molecular transport across human skin for drug delivery and analyte acquisition is of widespread interest. Creation of microconduits through the stratum corneum and epidermis is achieved by stochastic scissioning events localized to typically 250 μm diameter areas of human skin in vivo.     

Immunization with Porphyromonas gingivalis capsular polysaccharide prevents P. gingivalis-elicited oral bone loss in a murine model

The capsular polysaccharide (CPS) of the periodontal pathogen Porphyromonas gingivalis is an important virulence factor for this organism. We purified P. gingivalis CPS, immunized mice with this antigen, and assessed the vaccine potential of P. gingivalis CPS by using the murine oral challenge model. Animals immunized with P. gingivalis CPS developed elevated levels of immunoglobulin M (IgM) and IgG in serum that reacted with whole P. gingivalis organisms. The mice immunized with P. gingivalis CPS were protected from P. gingivalis-elicited oral bone loss. These data demonstrate that P. gingivalis CPS is a vaccine candidate for prevention of P. gingivalis-elicited oral bone loss.     

Single dipole localization: Some numerical aspects and a practical rejection criterion for the fitted parameters

Summary There have been a number of attempts in the last years to localize the generators of brain electromagnetic activity, considering one current dipole as the source model. Single Dipole Localization (SDL) requires the selection of an optimization algorithm (OA). General aspects related with the selection, implementation and evaluation of some of the OA employed for SDL are discussed in this paper. Specifically the performance of two algorithms, those of Hooke-Jeeves and Levenberg-Marquardt, are tested by simulations. Suggestions for including restrictions to the dipole position and comments about some commonly used measures of the goodness of fit are given. Examples of erroneous implementations of these algorithms are also illustrated. A simple graphic rejection criterion, which can be easily used by inexperienced researchers, is introduced and tested in noisy and noise free simulations.The authors are grateful to Roberto D. Pascual Marqui for programming the Hooke-Jeeves algorithm.     

Bacterial colonization of the uterine cervix and success rate in assisted reproduction: results of a prospective survey

BACKGROUND: Overgrowth of bacteria in the birth canal is associated with an increased risk of late miscarriage, preterm labour, post-partum endometritis and low birthweight. Conception rates in assisted reproduction treatments (ART) remain frustratingly low. We examined whether the nature of bacterial flora, found in the uterine cervical canal at embryo transfer, is associated with the rate of conception in ART. METHODS: We sampled for bacteriological culture the cervical canal of 204 patients who underwent embryo transfer. Of these, 139 (68%) were of fresh embryos, following recent vaginal oocyte retrieval and prophylactic antibiotic therapy, and 65 (32%) of frozen-thawed embryos, without any vaginal intervention in the preceding days. Bacteriological work-up included identification, colony count and antibiotic susceptibility profile. Conception was correlated with bacterial type and colony count. RESULTS: In 75 patients (36.8%) sterile cervical cultures or lactobacillus were recorded. Of these 75 patients, 23 (30.7%) conceived, whereas among the 129 in whom any pathogenic micro-organism was recovered only 21 (16.3%) conceived (P = 0.002). No difference in colonization was found between women who underwent frozen-thawed versus fresh embryo transfer (57 and 67% respectively). Any Gram-negative colonization was associated with no conception. All Gram-positive, and 90% of the Gram-negative bacteria, were sensitive to augmentin. CONCLUSIONS: Failure to conceive in ART is significantly associated with bacterial colonization of the uterine cervix.     

Peroxisome proliferator-activated receptor-α and liver cancer: where do we stand?

The peroxisome proliferator-activated receptor- (PPAR), first identified in 1990 as a member of the nuclear receptor superfamily, has a central role in the regulation of numerous target genes encoding proteins that modulate fatty acid transport and catabolism. PPAR is the molecular target for the widely prescribed lipid-lowering fibrate drugs and the diverse class of chemicals collectively referred to as peroxisome proliferators. The lipid-lowering function of PPAR occurs across a number of mammalian species, thus demonstrating the essential role of this nuclear receptor in lipid homeostasis. In contrast, prolonged administration of PPAR agonists causes hepatocarcinogenesis, specifically in rats and mice, indicating that PPAR also mediates this effect. There is no strong evidence that the low-affinity fibrate ligands are associated with cancer in humans, but it still remains a possibility that chronic activation with high-affinity ligands could be carcinogenic in humans. It is now established that the species difference between rodents and humans in response to peroxisome proliferators is due in part to PPAR. The cascade of molecular events leading to liver cancer in rodents involves hepatocyte proliferation and oxidative stress, but the PPAR target genes that mediate this response are unknown. This review focuses on the current understanding of the role of PPAR in hepatocarcinogenesis and identifies future research directions that should be taken to delineate the mechanisms underlying PPAR agonist-induced hepatocarcinogenesis.     

Distinct reovirus-like agents associated with acute infantile gastroenteritis.

Human reovirus-like particles were found by electron microscopy in the stools of 25% of 71 infants and young children hospitalized with acute gastroenteritis in Mexico between December 1976 and April 1977. The virus was also identified by the electrophoresis patterns of its ribonucleic acid upon disruption of partially purified particles. This technique is as reliable as electron microscopy but less laborious, and could become a routine diagnostic procedure. The electrophoretic patterns of viral ribonucleic acid from different cases suggest that there are at least two different reovirus-like agents associated with infantile gastroenteritis.