Free light chain UV quantification compared with immunochemical measurement: How dimers and monomers may influence the results

Serum κ and λ free light chain (FLC) levels are important for the management of plasma cell disorders. Immunochemical measurements on automated platforms with different reagents occasionally return different results that make them not interchangeable. The reasons for this behaviour are not clear and it is not known which result is the most accurate. The aim of the study is to quantify naturally occurring FLCs with a reference method (UV absorbance) in a sample devoid of other sources of UV absorbance. This was possible on a particular urine sample containing only lambda FLC proteins, dialyzed to clear it from low molecular weight UV absorbing compounds. The sample was submitted to Fast Protein Liquid Chromatography separation with a size-exclusion column in order to separate the FLC monomers and dimers. FLCs were also measured with the Freelite and N Latex FLC methods and the results were compared. The results demonstrated that the amount of FLC calculated on the basis of UV absorbance was overestimated by both immunochemical methods, and that the amount measured by the two reagents was affected by the different proportions of dimers or monomers. The present findings may be useful for the comprehension of the immunochemical measurement of FLC

<b><i>Topoisomerase 1</i></b> Promoter Variants and Benefit from Irinotecan in Metastatic Colorectal Cancer Patients

Objective: Topoisomerase 1 (topo-1) is an important target for the treatment of metastatic colorectal cancer (CRC). The aim of the present study was to evaluate the correlation between topo-1 single-nucleotide polymorphisms (SNPs) and clinical outcome in metastatic CRC (mCRC) patients. Methods: With the use of specific software (PROMO 3.0), we performed an in silico analysis of topo-1 promoter SNPs; the rs6072249 and rs34282819 SNPs were included in the study. DNA was extracted from 105 mCRC patients treated with FOLFIRI ± bevacizumab in the first line. SNP genotyping was performed by real-time PCR. Genotypes were correlated with clinical parameters (objective response rate, progression-free survival, and overall survival). Results: No single genotype was significantly associated with clinical variables. The G allelic variant of rs6072249 topo-1 SNP is responsible for GC factor and X-box-binding protein transcription factor binding. The same allelic variant showed a nonsignificant trend toward a shorter progression-free survival (GG, 7.5 months; other genotypes, 9.3 months; HR 1.823, 95% CI 0.8904-3.734; p = 0.1). Conclusion: Further analyses are needed to confirm that the topo-1 SNP rs6072249 and transcription factor interaction could be a part of tools to predict clinical outcome in mCRC patients treated with irinotecan-based regimens

A kinetic study of gamma-glutamyltransferase (GGT)-mediated S-nitrosoglutathione catabolism.

S-Nitrosoglutathione (GSNO) is a nitric oxide (NO) donor compound which has been postulated to be involved in transport of NO in vivo. It is known that c-glutamyl transpeptidase (GGT) is one of the enzymes involved in the enzyme-mediated decomposition of GSNO, but no kinetics studies of the reaction GSNO-GGT are reported in literature. In this study we directly investigated the kinetics of GGT with respect to GSNO as a substrate and glycyl- glycine (GG) as acceptor co-substrate by spectrophotometry at 334 nm. GGT hydrolyses the c-glutamyl moiety of GSNO to give S-nitroso-cysteinylglycine (CGNO) and c-glutamyl-GG. However, as both the substrate GSNO and the first product CGNO absorb at 334 nm, we optimized an ancillary reaction coupled to the enzymatic reaction, based on the copper-mediated decomposition of CGNO yielding oxidized cysteinyl-glycine and NO. The ancillary reaction allowed us to study directly the GSNO/GGT kinetics by following the decrease of the characteristic absorbance of nitrosothiols at 334 nm. A Km of GGT for GSNO of 0.398 ± 31 mM was thus found, comparable with Km values reported for other c-glutamyl substrates of GGT

Unusual onset of a case of chronic recurrent multifocal osteomyelitis

Background: Chronic recurrent multifocal osteomyelitis (CRMO) is a rare condition that commonly affects the clavicle and pelvis. Case presentation: We report here a case a 12 years old girl with CRMO arising with recurrent episodes of left supraorbital headache, followed by the appearance of a periorbital dyschromia. Magnetic resonance imaging (MRI) of the skull and orbits revealed an important subacute inflammatory process. Few months after, the child presented a painful swelling of the left clavicle; the histological examination of the related biopsy allowed to establish the diagnosis of CRMO. Conclusion: CRMO presenting as acute headache involving neurocranium is rare; to our knowledge this is the first recognized case in the world literature. This pathological condition is frequently misdiagnosed as infection or neoplasm and needs a deep investigation for the differential diagnosis. The physical, laboratoristic and instrumental diagnostic investigations of the patient and the treatment employed are described in detail

CYTOCHROME P450 3A13 AND ENDOTHELIN JOINTLY MEDIATE DUCTUS ARTERIOSUS CONSTRICTION TO OXYGEN IN MICE

The fetal ductus arteriosus (DA) contracts to oxygen, and this feature, maturing through gestation, is considered important for its closure at birth. We have previously obtained evidence of the involvement of cytochrome P-450, possibly of the 3A subfamily (CYP3A), in oxygen sensing and have also identified endothelin (ET)-1 as the attendant effector for the contraction. Here, we examined comparatively wild-type (WT) and CYP3Anull (Cyp3a(-/-)) mice for direct validation of this concept. We found that the CYP3A subfamily is represented only by CYP3A13 in the WT DA. CYP3A13 was also detected in the DA by immunofluorescence microscopy, being primarily colocalized with the endoplasmic reticulum in both endothelial and muscle cells. However, a distinct signal was also evident in the plasma membrane. Isolated DAs from term WT animals developed a sustained contraction to oxygen with transient contractions superimposed. Conversely, no tonic response occurred in Cyp3a(-/-) DAs, whereas the phasic response persisted unabated. Oxygen did not contract the preterm WT DA but caused a full-fledged contraction after retinoic acid (RA) treatment. RA also promoted an oxygen contraction in the Cyp3a(-/-) DA. However, responses of RA-treated WT and Cyp3a(-/-) mice differed in that only the former abated with ET-1 suppression. This implies the existence of an alternative target for RA responsible for the oxygen-induced contraction in the absence of CYP3A13. In vivo, the DA was constricted in WT and Cyp3a(-/-) newborns, although with a tendency to be less narrowed in the mutant. We conclude that oxygen acts primarily through the complex CYP3A13 (sensor)/ET-1 (effector) and, in an accessory way, directly onto ET-1. However, even in the absence of CYP3A13, the DA may close postnatally thanks to the contribution of ET-1 and the likely involvement of compensating mechanism(s) identifiable with an alternative oxygen-sensing system and/or the withdrawal of relaxing influence(s) operating prenatally

Physical Investigation of the Potentially Hazardous Asteroid (144898) 2004 VD17

In this paper we present the observational campaign carried out at ESO NTT and VLT in April and May 2006 to investigate the nature and the structure of the Near Earth Object (144898) 2004 VD17. In spite of a great quantity of dynamical information, according to which it will have a close approach with the Earth in the next century, the physical properties of this asteroid are largely unknown. We performed visible and near--infrared photometry and spectroscopy, as well as polarimetric observations. Polarimetric and spectroscopic data allowed us to classify 2004 VD17 as an E-type asteroid. A good agreement was also found with the spectrum of the aubrite meteorite Mayo Belwa. On the basis of the polarimetric albedo (p_v=0.45) and of photometric data, we estimated a diameter of about 320 m and a rotational period of about 2 hours. The analysis of the results obtained by our complete survey have shown that (144898) 2004 VD17 is a peculiar NEO, since it is close to the breakup limits for fast rotator asteroids, as defined by Pravec and Harris (2000). These results suggest that a more robust structure must be expected, as a fractured monolith or a rubble pile in a "strength regime" (Holsapple 2002).Comment: 32 pages, 7 figure, paper accepted for publication in Icaru

Prospective qualitative and quantitative non-invasive evaluation of intestinal acute GVHD by contrast-enhanced ultrasound sonography.

Intestinal acute GVHD (I-aGVHD) is a life-threatening complication after allografting. Non-invasive bed-side procedures to evaluate extension and treatment response are still lacking. We hypothesized that, during I-aGVHD, contrast-enhanced ultrasound sonography (CEUS) could detect microcirculation changes (MVC) of the bowel wall (BW) and help to monitor treatment response. We prospectively employed CEUS in 83 consecutive patients. Of these, 14 patients with biopsy-proven intestinal GVHD (I-GVHD) were defined as the study group, whereas 16 patients with biopsy-proven stomach GVHD (U-GVHD) without intestinal symptoms, 6 normal volunteers and 4 patients with neutropenic enterocolitis were defined as the control group. All patients were evaluated with both standard ultrasonography (US) and CEUS at the onset of intestinal symptoms, during clinical follow-up and at flare of symptoms. Standard US revealed BW thickening of multiple intestinal segments, useful to determine the extension of GVHD. CEUS showed MVC, which correlated with GVHD activity, treatment response, and predicted flare of intestinal symptoms. US and CEUS findings were superimposable at diagnosis and in remission. CEUS was, however, more sensitive and specific to identify subclinical activity in patients with clinical relevant improvement. These findings were not observed in the control groups. CEUS is a non-invasive, easily reproducible bed-side tool useful to monitor I-aGVHD

Detección de anticuerpos séricos en toros inmunizados contra campylobacteriosis

En el presente ensayo se evaluó la repuesta sérica vacunando 169 toros jóvenes para carne, vírgenes de 12–24 meses de edad, inmunizados con vacunas a célula entera de Campylobacter fetus. El trabajo se realizó en cuatro establecimientos libres de campylobacteriosis de la provincia de Buenos Aires (Argentina). Se utilizaron tres vacunas comerciales nacionales, conteniendo cepas inactivadas de C. fetus y sus subspecies: vacunas A (n= 84), B (n=19) y C (n=58) (suspensión de C. fetus subsp. fetus, intermedius y venerealis, inactivadas con formol). Las vacunas A y B poseían como vehículo hidróxido de aluminio, la vacuna C presentaba adyuvante oleoso. También se utilizó una vacuna experimental (D) (n=8) dual, oleosa, elaborada por el INTA Balcarce, conteniendo antígenos inactivados de C. fetus subsp. fetus, C. fetus subsp. venerealis y Tritrichomonas foetus. Todas las vacunas se aplicaron por vía subcutánea, dos dosis con intervalo de 21 días. Los animales se sangraron a los días 0, 21, 42, 93 y 123 post primera dosis vacunal (DPV). Los sueros fueron procesados por ELISA indirecto. Los valores de absorbancia expresados en densidad óptica fueron transformados a valores ELISA (VE). La vacuna experimental dual D demostró un incremento importante en los VE de los anticuerpos séricos con respecto las vacunas A, B y C (p&lt; 0,05). Las vacunas B y C tuvieron mejor respuesta a los 21 y 42 días DPV respecto a la vacuna A (p&lt; 0,05). A los 93 DPV, los animales del grupo C tuvieron un débil incremento de VE en comparación con aquellos animales de los grupos A y B (p&lt; 0,05). El desempeño general tendió a ser mayor en los animales inmunizados con la vacuna experimental que aquellos del grupo de vacunas comerciales. Se observaron VE muy bajos en general en las vacunas comerciales utilizadas. El método ELISA fue adecuado para la evaluación de la respuesta inmune sistémica en los toros vacunados.