Knock‐down of HEXA and HEXB genes correlate with the absence of the immunostimulatory function of HSC‐derived dendritic cells

In an attempt to investigate whether the genetic defect in the HEXA and HEXB genes (which causes the absence of the lysosomal β‐N‐acetyl‐hexosaminidase), are related to the wide inflammation in GM2 gangliosidoses (Tay‐Sachs and Sandhoff disease), we have chosen the dendritic cells (DCs) as a study model. Using the RNA interference approach, we generated an in vitro model of HEXs knock‐down immunogenic DCs (i‐DCs) from CD34⁺‐haemopoietic stem cells (CD34⁺‐HSCs), thus mimicking the Tay‐Sachs (HEXA?/?) and Sandhoff (HEXB?/?) cells. We showed that the absence of β‐N‐acetyl‐hexosaminidase activity does not alter the differentiation of i‐DCs from HSCs, but it is critical for the activation of CD4⁺T cells because knock‐down of HEXA or HEXB gene causes a loss of function of i‐DCs. Notably, the silencing of the HEXA gene had a stronger immune inhibitory effect, thereby indicating a major involvement of β‐N‐acetyl‐hexosaminidase A isoenzyme within this mechanism. Copyright © 2011 John Wiley & Sons, Ltd.     

Plasma renin activity and urinary kallikrein excretion in lead-exposed workers as related to hypertension and nephropathy

Urinary kallikrein activity and plasma renin activity (PRA) in supine and erect positions were determined in 22 men occupationally exposed to lead. In eight of them, suffering from hypertension and/or nephropathy, urinary kallikrein activity was low or absent, while PRA was normal or reduced. The other fourteen non-symptomatic workers showed normal or reduced urinary kallikrein activity and variable PRA. Urinary kallikrein activity of lead-exposed workers was significantly correlated with upright PRA.The relation between lead-exposure and essential hypertension is discussed.     

Comparative growth of the thermal alga Cyanidium caldarium on nitrate and ammonia at different temperatures

Summary The growth of Cyanidium caldarium on nitrate and ammonia as nitrogen sources was studied at different temperatures from 21 to 54°C.Algal growth occurred at temperatures of 24° C or above when ammonia was the nitrogen source, whereas with nitrate, growth occurred at 30° C or above. The optimum and the maximum growth temperatures were 45 and 54° C respectively on both substrates.Arrhenius plots show that the logarithm of the growth rate is not linear with the reciprocal of absolute temperature, but exhibit sharply defined breaks at 30° C on ammonia and at 40° C on nitrate.It is assumed that below 40° C, when Cyanidium grows on nitrate, the utilization of this substrate represents the master reaction which controls the growth rate of the alga.     

Effects of pentobarbital on serum levels of LH and FSH, in normal conditions and after GnRH stimulation: preliminary report

The authors have tested serum levels of LH and FSH in healthy males anaesthetized with 20 mg/Kg body weight of pentobarbital, and stimulation test with GnRH. LH has reached levels higher than the stimulation test has been practised in the anaesthetized volunteers than in subjects awakes. The AA. have also showed that there is no effect on psychic stress and of atropine, used as preanaesthetic drug, on serum levels of the pituitary hormones. At the end, PB has no effect on serum levels of FSH.     

HPLC method to characterize cyanogen bromide collagen fractions containing pyridinoline groups.

The HPLC method here described allows to separate CNBr collagen peptides within 2.5 h by reversed phase and gradient elution. The method is useful to determine both peptide bond and pyridinoline groups by absorbance spectophotometry. The fractions can be recovered and then submitted to other characterization techniques.     

Isothermal and non-isothermal bioreactors in the detoxification of waste waters polluted by aromatic compounds by means of immobilised laccase from Rhus vernicifera

Laccase from Rhus vernicifera was immobilised on a nylon membrane chemically grafted with glycidyl methacrylate (GMA). Hexamethylenediamine (HMDA) and glutaraldehyde (GLU) were used as spacer and bifunctional coupling agent, respectively. Quinol was used as substrate.

To know how the immobilisation procedures affected the enzyme reaction rate the catalytic behaviour of soluble and insoluble laccase was studied under isothermal conditions as a function of pH, temperature and substrate concentration. From these studies, two main singularities emerged from the experimental data: (i) the narrower pH-activity profile of the insoluble enzyme in comparison to that of the soluble counterpart; (ii) the increase of the affinity of the immobilised enzyme for its substrate.

The behaviour of the catalytic membrane was also studied in a non-isothermal bioreactor as a function of substrate concentration and size of the applied transmembrane temperature difference. It was found that, under non-isothermal conditions and keeping constant the average temperature of the bioreactor, the enzyme reaction rate linearly increases with the increase of the temperature difference. These results have been discussed in the frame of reference of the process of thermodialysis driving thermodiffusive transmembrane substrate fluxes, which add to the diffusive ones.

The advantages of the catalytic process carried out under non-isothermal conditions have been thrown in relief through the evaluation of the reduction of the production times and of the percentage increases of the enzyme activity.     

IGFBP-3 and IGFBP-10 (CYR61) up-regulation during the development of Barrett's oesophagus and associated oesophageal adenocarcinoma: potential biomarkers of disease risk.

Dys-regulation of the insulin-like growth factor (IGF) system increases the risk of a number of malignancies. The aim of this study was to investigate the role of members of the IGF binding protein (IGFBP) superfamily in the development of oesophageal adenocarcinoma (EAC) and their possible use as markers of disease risk. Expression of IGFBP-2, IGFBP-3, IGFBP-4, and IGFBP-10/CYR61 was assessed using Real-Time-polymerase chain reaction (PCR) and immunohistochemistry in oesophageal tissues from Barrett's oesophagus (BE) patients with and without associated EAC, and in control subjects. IGFBP-3, IGFBP-4, and IGFBP-10/CYR61 mRNA levels were up-regulated in Barrett's (n=17) and tumour tissue of EAC patients (n=18) compared with normal tissue of control subjects without BE or EAC (n=18) (p<0.001). Over-expression of IGFBP-3 and IGFBP-10/CYR61 proteins was observed in Barrett's, dysplastic and tumour tissue of EAC cases (n=47 for IGFBP-10; n=39 for IGFBP-3) compared with adjacent normal epithelium (p<0.050). Notably, IGFBP-3, IGFBP-4, and IGFBP-10/CYR61 expression in Barrett's tissue of EAC cases (n=17) was significantly (p<0.001) higher than in Barrett's tissue of BE patients with no sign of progression to cancer (n=15). Overall, the results suggest that members of the IGFBP superfamily are up-regulated during oesophageal carcinogenesis and merit further investigation as markers of EAC risk.