MEMRI and tumors: a method for the evaluation of the contribution of Mn(II) ions in the extracellular compartment

The purpose of the work was to set‐up a simple method to evaluate the contribution of Mn²⁺ ions in the intra‐ and extracellular tumor compartments in a MEMRI experiment. This task has been tackled by “silencing” the relaxation enhancement arising from Mn²⁺ ions in the extracellular space. In vitro relaxometric measurements allowed assessment of the sequestering activity of DO2A (1,4,7,10‐tetraazacyclododecane‐1,7‐diacetic acid) towards Mn²⁺ ions, as the addition of Ca‐DO2A to a solution of MnCl₂ causes a drop of relaxivity upon the formation of the highly stable and low‐relaxivity Mn‐DO2A. It has been proved that the sequestering ability of DO2A towards Mn²⁺ ions is also fully effective in the presence of serum albumin. Moreover, it has been shown that Mn‐DO2A does not enter cell membranes, nor does the presence of Ca‐DO2A in the extracellular space prompt migration of Mn ions from the intracellular compartment. On this basis the in vivo, instantaneous, drop in SE% (percent signal enhancement) in T₁‐weighted images is taken as evidence of the sequestration of extracellular Mn²⁺ ions upon addition of Ca‐DO2A. By applying the method to B16F10 tumor bearing mice, T₁ decrease is readily detected in the tumor region, whereas a negligible change in SE% is observed in kidneys, liver and muscle. The relaxometric MRI results have been validated by ICP‐MS measurements. Copyright © 2015 John Wiley & Sons, Ltd.     

Buprenorphine in balanced anesthesia

Buprenorphine was administered for the balanced general anaesthesia in 300 patients during several operations. The results obtained, the good features of analgesia, morphine-like, the absence of cardiovascular and respiratory side effects are presented.     

Dog-ears: a useful artifice in the closure of extensive wounds

BACKGROUND: Wounds too extensive to permit primary repair by suturing can be closed using a skin graft or skin flap and the choice of method depends on a series of factors. Practice and personal experience play a role, as well as the characteristics of the lesion and its site. Each case poses special problems, so it is not possible to establish firm rules. OBJECTIVE: To present a surgical technique for the closure of a circular defect located on the dorsum of the foot, which resulted from the extirpation of a melanoma by surgery. METHODS: In this case we used a mixed procedure consisting of skin flaps and a double full-thickness skin graft. For this operation we used dog-ears resulting from the circular exeresis performed for the extirpation of the tumour. RESULTS: The reconstructive results were good and there were no postoperative complications. CONCLUSIONS: We consider this procedure particularly suitable for certain anatomical sites, such as the dorsum of the foot, where the skin is less elastic and thinner; and for patients who are not willing to accept skin removal from other sites or for flaps to be made with more invasive operations and more constructed scars.     

Critical evaluation of serum thyroglobulin (Tg) levels during thyroid hormone suppression therapy versus Tg levels after hormone withdrawal and total body scan: results in 291 patients with thyroid cancer

Serum thyroglobulin (Tg) levels were measured during thyroid-hormone suppressive therapy in 291 patients who had been treated for well-differentiated thyroid carcinoma. The findings were compared with those of a subsequent total body scan (TBS) and with Tg levels measured after thyroid-hormone withdrawal. Of the patients with low Tg levels during suppressive therapy, 91.6% were subsequently shown to be free of disease or to have only remnants in the thyroid bed, whereas 8.4% had metastases (false-negative). Of the patients with false-negative findings, 89.3% had nodal metastases; 60.8% of the patients with nodal metastases exhibiting radioiodine uptake and only 23% of those with nonfunctioning nodal metastases had low Tg levels during suppression therapy. After thyroid-hormone withdrawal, all but 1 of the patients with nodal metastases had high Tg levels. All but 2 of the patients with distant metastases had high Tg levels during suppression therapy; 1 of these 2 patients exhibited high Tg levels after T3 withdrawal. No differences between Tg levels in patients with functioning and non-functioning metastases were found. Our study indicates that Tg assays performed during suppressive therapy have a fairly good predictive value and can be used as a general guide in the follow-up of thyroid cancer. However, since most differentiated metastases produce Tg only when stimulated by thyroid-stimulating hormone, measurements of Tg levels after the discontinuation of suppressive therapy would also seem to be necessary.     

Development of a real-time PCR assay for detection of Plasmodium falciparum, Plasmodium vivax, and Plasmodium ovale for routine clinical diagnosis

A TaqMan-based real-time PCR qualitative assay for the detection of three species of malaria parasites-Plasmodium falciparum, P. ovale, and P. vivax-was devised and evaluated using 122 whole-blood samples from patients who had traveled to areas where malaria is endemic and who presented with malaria-like symptoms and fever. The assay was compared to conventional microscopy and to an established nested-PCR assay. The specificity of the new assay was confirmed by sequencing the PCR products from all the positive samples and by the lack of cross-reactivity with Toxoplasma gondii and Leishmania infantum DNA. Real-time PCR assay showed a detection limit (analytical sensitivity) of 0.7, 4, and 1.5 parasites/ micro l for P. falciparum, P. vivax, and P. ovale, respectively. Real-time PCR, like nested PCR, brought to light errors in the species identification by microscopic examination and revealed the presence of mixed infections (P. falciparum plus P. ovale). Real-time PCR can yield results within 2 h, does not require post-PCR processing, reduces sample handling, and minimizes the risks of contamination. The assay can therefore be easily implemented in routine diagnostic malaria tests. Future studies are warranted to investigate the clinical value of this technique.     

Analysis of susceptibility of mature human T lymphocytes to dexamethasone-induced apoptosis

We present evidence that dexamethasone (Dex), a synthetic glucocorticosteroid, causes apoptosis in mature human T cells, similarly to what has been reported for murine T lymphocytes. Human T cell clones and short-term activated T lymphocytes treated with Dex show the characteristic pattern of apoptotic cells, such as hypodiploid nuclei, chromatin condensation and DNA fragmentation into oligonucleosomal fragments. However, Dex susceptibility of T cells to apoptosis is cell cycle-dependent. The progression in the proliferative cell cycle (G1 versus S) rescues Dex-treated T cells from apoptosis. Moreover, occupancy of the T cell receptor reverses Dex-induced apoptotic phenomena. These observations suggest that glucocorticoids contribute to the regulation of the proliferative or the suicidal response of antigen-activated human T cells.     

Mutational diversity and hot spots in the alpha-sarcoglycan gene in autosomal recessive muscular dystrophy (LGMD2D).

Sarcoglycanopathies are a genetically heterogeneous group of autosomal recessive muscular dystrophies in which the primary defect may reside in any of the genes coding for the different partners of the sarcolemmal sarcoglycan (SG) complex: the alpha-SG (LGMD2D at 17q21.2), the beta-SG (LGMD2E at 4q12), the gamma-SG (LGMD2C at 13q12), and the delta-SG (LGMD2F at 5q33). We report a series of 20 new unrelated families with 14 different mutations in the alpha-SG gene. Along with the mutations that we previously reported this brings our cohort of patients with alpha-sarcoglycanopathy to a total of 31 unrelated patients, carrying 25 different mutations. The missense mutations reside in the extracellular domain of the protein. Five of 15 missense mutations, carried by unrelated subjects on different haplotype backgrounds and of widespread geographical origins, account for 58% of the mutated chromosomes, with a striking prevalence of the R77C substitution (32%). The severity of the disease varies strikingly and correlates at least in part with the amount of residual protein and the type of mutation. The recurrent R284C substitution is associated with a benign disease course.