Rapid single-step FCGR3A genotyping based on SYBR Green I fluorescence in real-time multiplex allele-specific PCR

A polymorphism at position 559 in the cDNA of the FCGR3A gene encoding the FcgammaRIIIa generates two allotypes with either a valine (Val) or a phenylalanine (Phe) at amino acid position 158. This polymorphism is of major importance in immunopathology and in pharmacogenetics, especially monoclonal antibody treatments. In this study, we report a single-step and single-tube method for FCGR3A-158V/F genotyping by real-time multiplex allele-specific PCR and melting curve analysis in the presence of SYBR Green I fluorescent dye. Results obtained from 113 samples showed 100% accuracy compared to those obtained with a conventional PCR-based allele-specific restriction assay. Although this method requires expensive equipment, it is inexpensive in terms of consumables. It is also very rapid, reliable and suitable for large-scale analysis.     

Is angiotensin II a proliferative factor of cardiac fibroblasts?

Angiotensin II has been implicated as an important factor in cardiac remodeling, particularly in the development of pathological left ventricular hypertrophy. It is generally assumed that angiotensin II is able to alter the phenotype of cardiac myocytes and fibroblasts, and several experiments have suggested that this peptide can particularly affect the proliferation of cardiac fibroblasts. However, a review of the published results indicates that there is no evidence that angiotensin II can directly trigger mitogenesis through activation of the cyclin-dependent pathway. The observed proliferative effect might well be caused by stimulation of the synthesis of growth or inflammatory substances like platelet-derived growth factor and cytokines, by integrin activation due to secreted extracellular matrix proteins, or by a combination of these mechanisms. Angiotensin II thus appears to differentiate cardiac fibroblasts into a growth substance-secreting phenotype.     

Bladder involvement in scleroderma: report of 3 cases

Lower urinary tract involvement is the common manifestation of systemic sclerosis. It may represent a troublesome disturbance affecting the quality of life in systemic sclerosis patients. We report three patients with progressive systemic sclerosis who manifested pathologic abnormalities in the urinary bladder. The first patient manifested a recurrent macroscopic hematuria requiring transfusions. Both other patients developed functional and urodynamic abnormalities. With a review of the literature on this involvement of the bladder in systemic sclerosis we try to more understand urinary symptoms and urodynamics features of vesical involvement in systemic sclerosis patients.     

Critical clinicopathologic analysis of 23 cases of fine-needle breast sampling initially recorded as false-positive. The 44-year experience of the Institut Curie

BACKGROUND: Because false-positive cytologic diagnoses in breast tumors are rare, few cases have been reported, although their consequences may be highly detrimental to the patient. The authors report the Institut Curie's experience, by using a multidisciplinary approach. METHODS: Of 9334 benign breast tumors examined preoperatively for cytologic diagnosis by fine-needle sampling (FNS), the 23 (0.25%) FNS cases considered to be false-positive were retrospectively reviewed and analyzed. RESULTS: Tumors were situated close to the nipple in 7 cases and away from the nipple in 16 cases. Tumor stage was T0 for 1 case, T1 for 18 cases, and T2 for 4 cases. Radiologically, six tumors were classified as malignant, seven as indeterminate or suspicious, and nine as benign. Three of six tumors studied by flow cytometry were DNA aneuploid. Based on a multidisciplinary clinicopathologic review, 20 FNS cases were finally classified as false-positive, and the remaining 3 tumors with malignant FNS and subsequent benign histology were classified as true-positive, because local and/or metastatic progression was observed in the short term. CONCLUSIONS: The authors' review suggests two categories of false-positive cases: the first in which cytologic benign patterns are overdiagnosed, and the second in which atypical morphologic criteria were present. Nevertheless, as shown by the malignant course in three cases, patients with malignant preoperative FNS and corresponding benign histology always require close clinical follow-up. Finally, surgical overtreatment rate could be decreased if all radiologically benign tumors with positive/suspicious FNS were subject to intraoperative frozen section examination.     

Comparative effect of ANF and various diuretics on isolated nephron segments

The effect of the synthetic form of ANF 0.1 to 10 microgram/ml (peptide 101-126), a diuretic and natriuretic peptide isolated from rat heart atria, on the metabolism of dog and rat kidney tubules was studied in vitro and compared to that of furosemide (0.1 to 1 mM), hydrochlorothiazide (0.5 mM) or amiloride (0.1 mM). In order to pinpoint eventual site(s) of ANF action along the nephron, proximal tubules, thick ascending limbs and papillary collecting ducts were isolated from dog kidneys as well as proximal tubules from rat kidneys. The substrate uptake (O2, lactate, glutamine, glucose) and production of metabolites (glutamate, ammonium, alanine, glucose) by these nephron segments were measured in absence or presence of the diuretic agents or the vehicle for ANF (acetate 1 mM). The total ATP turnover and the contribution of identified metabolic pathways for this turnover was calculated. It was expected that a molecule with diuretic properties reducing the permeability of cell membranes to NaCl would secondarily reduce the Na-K-ATPase activity, and therefore the oxygen and substrate utilization by affected cells. It was shown: that each nephron segment used presented the expected specific metabolic characteristics; that furosemide markedly inhibits the oxidative metabolism of thick ascending limbs; that acetate (the vehicle used for ANF) displaces the oxidation of glutamine and lactate in nephron segments with aerobic metabolism; that ANF had no effect on the metabolism of the studied segments despite the presence of specific c'GMP-generating receptors in the distal nephron. It is concluded that ANF must exert its natriuretic effect by a mechanism different from that of classical diuretics.