Glycolipid and protein profiles of normal and Trypanosoma cruzi infected heart muscle cells.

Using Triton X-114, glycolipids and proteins were extracted from heart muscle cells (HMC) infected with Trypanosoma cruzi clone Dm28c and from uninfected HMC, and analysed by SDS-PAGE and high-performance thin-layer chromatography (HPTLC). Two major differences were observed: (a) two proteins with a molecular mass of 92 kDa and 69 kDa were present in the uninfected cells but absent from the infected cells and (b) a 70-90 kDa protein band was detected only in parasitized cells. These differences would seem to constitute alterations taking place during the process of cell recognition and/or parasite interiorization. No differences were observed in the respective glycolipid compositions, of control and infected cells analysed by HPTLC. A glycolipid with the same mobility as the neutral glycolipid isolated from epimastigotes of T. cruzi was detected in the uninfected cells. This finding may lend support to the previously described hypothesis that molecular mimicry is implicated in the cardioneuropathology of Chagas' disease.     

Type 2 diabetes: assessment of endothelial lesion and fibrinolytic system markers.

This study aimed to investigate whether endothelial cells are damaged and to evaluate fibrinolytic system function in patients with type 2 diabetes. For this proposal, plasma levels of von Willebrand factor (an endothelial marker of injury), homocysteine (an inductor of endothelial injury), D-dimer (a marker of coagulation cascade activation) and plasminogen activator inhibitor-1 (a fibrinolysis marker) were measured in individuals with both type 2 diabetes and high blood pressure, with type 2 diabetes, with high blood pressure and in healthy control individuals. No significant differences among groups were observed for von Willebrand factor and homocysteine plasma levels. The type 2 diabetes and high blood pressure group presented a significant difference to the other groups for D-dimer and also presented high values for plasminogen activator inhibitor-1. The high blood pressure group and type 2 diabetes group presented separately higher values of plasminogen activator inhibitor-1 compared with the control group. High levels of D-dimer and plasminogen activator inhibitor-1 in patients with type 2 diabetes and high blood pressure with normoalbuminuria therefore indicate a state of hypercoagulability and hypofibrinolysis, despite no evident microvascular injury supported by normal levels of von Willebrand factor and homocysteine.     

Diagnostic accuracy of fetal renal pelvis anteroposterior diameter as a predictor of uropathy: a prospective study.

OBJECTIVE: The purpose of this study was to assess the accuracy of prenatal ultrasound measurement of anteroposterior renal pelvis diameter (APD) to discriminate between significant uropathy and idiopathic renal pelvis dilatation. METHODS: One-hundred-and-three neonates who were found to have fetal renal pelvis dilatation, defined as presence of an APD > or = 5 mm, underwent systematic investigation for uropathies and were prospectively followed. An ultrasound scan was performed after the first week of postnatal life and all infants underwent a voiding cystourethrogram. Neonates with an APD larger than 10 mm underwent renal scintigraphy. Ultrasound scans, clinical examination and laboratory reviews were scheduled at 6-month intervals. Receiver-operating characteristics (ROC) curves were constructed to determine the best cut-offs for APD to identify renal units with significant uropathy as well as those requiring surgical intervention. Significant uropathy was defined as the presence of well-established urinary tract abnormalities or when there was abnormal renal scintigraphy. RESULTS: The estimated area under the curve for APD was 0.900 (95% CI, 0.841-0.942) indicating excellent power to discriminate between idiopathic pelvis dilatation and significant uropathy. The sensitivity and specificity for the 7.5 mm cut-off point were 97.9% and 40.6%, respectively. To identify infants who required surgical intervention, the calculated area under the curve was 0.953 (95% CI, 0.908-0.980). CONCLUSION: Our results suggest that measurement of APD is an excellent test to identify fetuses with significant uropathy, as well as those requiring postnatal intervention.     

Acute Antibody-Mediated Rejection Following Heart Transplantation

Acute antibody-mediated rejection (AMR) in heart transplantation is often associated with hemodynamic compromise, and is associated with increased mortality and development of accelerated transplant coronary artery disease (TCAD). The diagnosis of AMR has historically been controversial and outcomes with aggressive immunosuppressive therapy including plasmapheresis and cyclophosphamide are poor. Advances in diagnostic techniques like the demonstration of immunopathologic evidence for antibody-mediated rejection by deposition of the complement split product C4d in tissue and detection of anti-HLA antibodies by flow cytometry will assist in further characterizing AMR. Immunosuppression targeting B-lymphocytes and use of m-TOR inhibitors to alter the predilection to develop TCAD and improve survival in AMR remains to be proven.     

Frequent ongoing T-cell receptor rearrangements in childhood B- precursor acute lymphoblastic leukemia: implications for monitoring minimal residual disease

Crosslineage T-cell receptor delta (TCR delta) rearrangements are widely used as tumor markers for the follow up of minimal residual disease in childhood B-precursor acute lymphoblastic leukemia (ALL) by polymerase chain reaction (PCR). The major drawback of this approach is the risk of false-negative results due to clonal evolution. We investigated the stability of V delta 2D delta 3 rearrangements in a group of 56 childhood B-precursor ALL patients by PCR and Southern blot analysis. At the PCR level, V delta 2D delta 3-to-J alpha rearranged subclones (one pathway for secondary TCR delta recombination) were demonstrated in 85.2% of V delta 2D delta 3-positive patients tested, which showed that small subclones are present in the large majority of patients despite apparently monoclonal TCR delta Southern blot patterns. Sequence analysis of V delta 2D delta 3J alpha rearrangements showed a biased J alpha gene usage, with HAPO5 and J alpha F in 26 of 32 and 6 of 32 clones, respectively. Comparison of V delta 2D delta 3 rearrangement status between diagnosis and first relapse showed differences in seven of eight patients studied. In contrast, from first relapse onward, no clonal changes were observed in six patients studied. To investigate the occurrence of crosslineage TCR delta rearrangements in normal B and T cells, fluorescence-activated cell sorter-sorted peripheral blood CD19+/CD3- and CD19-/CD3+ cell populations from three healthy donors were analyzed. V delta 2D delta 3 rearrangements were detected at low frequencies in both B and T cells, which suggests that V delta 2-to-D delta 3 joining also occurs during normal B-cell differentiation. A model for crosslineage TCR delta rearrangements in B-precursor ALL is deduced that explains the observed clonal changes between diagnosis and relapse and is compatible with multistep leukemogenesis of B-precursor ALL.     

P64Shoe contact dermatitis

Because of its numerous etiologies, foot dermatitis can be difficult to diagnose despite of thorough history and physical examination. The differential diagnoses are challenging and include allergic contact dermatitis, dyshidrosis, juvenile plantar dermatosis, atopic dermatitis, lichen planus.
Methods:  In the Cutaneous Allergology Unit of the Department of Dermatovenereology of Pedro Hispano Hospital, between 1999 and 2003, 804 patients were observed with suspicion of contact dermatitis. In 9,3% there was a history of recurrent foot dermatitis. All these patients were patch tested with GPEDC standard and shoe series and shoe fragments.
Results:  Twenty‐five patients revealed shoe contact dermatitis with positive reactions to the series tested, with an average age of 40 years. The dermatitis involved the dorsal aspect of the foot in 20 patients and the volar aspect in 8, the lateral aspects in 4, typically sparing the instep and flexural creases of the toes. The most common allergen were para‐tertiary‐butylphenol formaldehyde resin – PTBFR (12 patients‐48%), mercapto mix (7 patients‐28%), potassium dichromate (24%), mercaptobenzothiazole (24%).
Conclusions:  The most common causes of shoe contact dermatitis were glues, followed by rubber components and chromated leather. As in other studies the most common allergen in shoe dermatitis in Portugal is PTBFR in neoprene adhesives. Rubber components (accelerators) were the second more frequent allergen in this study, probably a direct result of improved fixation of chrome and a change in footwear style and chrome sensitivity explains leather allergy. Other causes of foot contact dermatitis are iatrogenic complication, clothing (socks), cosmetics, adhesive tape and professional etiology.     

Schistosoma mansoni: impairment of the cell-mediated immune response in mice.

Skin-graft rejection in mice experimentally infected with Schistosoma mansoni is delayed when grafting is performed 60 days after the infection. In mice infected 30 days prior to the grafting, the grafts were rejected at the same time both in infected and in control animals. This observation indicates that impairment of cell-mediated immune response occurs in mice with mature S. mansoni infections.     

Salivary immunoglobulins in a patient with IgA deficiency

The saliva of an individual with selective IgA deficiency was found to contain IgG and IgM, with some of the IgM linked to secretory component. Some specimens showed evidence of low molecular weight immunoglobulin fragments, presumed to be the result of proteolysis.