A new T677C mutation of the aspartoacylase gene encodes for a protein with no enzymatic activity

ObjectiveTo verify the effect of and to date the unknown T677C mutation of the human N-acetylaspartoacylase (hASPA) gene on the function of the mutated enzyme.Design and methodsWild type and I226T-mutated proteins were expressed and purified from a transformed Escherichia coli colony. Enzymatic activities were measured in the presence of varying substrate concentrations.ResultsWhilst kinetic parameters of wild type hASPA were in line with data in literature, I226T-mutated hASPA showed no enzymatic activity.ConclusionData indicated that this new mutation might be responsible in homozygosis for the phenotype corresponding to Canavan disease.     

Comparing fibrin sealant with staples for mesh fixation in laparoscopic transabdominal hernia repair: a case control-study

Background

Laparoscopic hernia repair is not as popular as cholecystectomy. We have performed more than 3,000 laparoscopic herniorrhaphies using the trans-abdominal (TAPP) technique. To prevent recurrences we fix the polypropylene mesh with staples. The use of fibrin glue for graft fixation is a possible alternative.

Methods

We have performed 3,130 laparoscopic hernia repairs over 14 years. For mesh fixation we used titanium clips and observed a small number of complications. In July 2003 we started using fibrin glue (Tissucol®). The purpose of this retrospective longitudinal study was to evaluate if the use of fibrin sealant was as safe and effective as conventional stapling and if there were differences in post-operative pain, complications and recurrences.

Results

From July 2003 to June 2006 we performed 823 laparoscopic herniorrhaphies. Fibrin glue (Tissucol®) was used in 88 cases. Two homogeneous groups of 68 patients (83 cases) treated with fibrin glue and 68 patients (87 cases) where the mesh was fixed with staples, were compared. Patients with relevant associated diseases or large inguino-scrotal hernias were excluded. Operative times were longer in the group treated with fibrin glue with a mean of 35 minutes (range 22–65 mins) compared to the group treated with staples (25 minutes, range 14–50 mins). The time of hospital stay was the same (24 hours). Post-operative complications, that were more frequent in the stapled group, included trocar site pain, hematomas, intra-operative bleedings and incisional hernias. No significant difference was observed concerning seromas, chronic pain and recurrence rate.

Conclusions

Less post-operative pain, and a faster return to usual activities are the main advantages of laparoscopic repair compared to the traditional approach. The use of fibrin sealant reduces in our experience the risk of post- and intra-operative complications such as bleeding and incisional hernia; recurrence rates are similar, but the operative time is longer.

     

A retrospective study of the efficacy of JAK inhibitors or abatacept on rheumatoid arthritis-interstitial lung disease

Inflammopharmacology - To examine the effectiveness of Janus-kinase inhibitors (JAKis) or abatacept (ABA) in patients with rheumatoid arthritis-interstitial lung disease (RA-ILD). Patients with...     

Thalamocortical dysrhythmia and the thalamic reticular nucleus in behaving rats.

OBJECTIVES: The aim of this study was to investigate the effects of bilateral chemical lesion of the rostral pole of the thalamic reticular nucleus on EEG activities in freely moving rats applying quantitative analysis and brain mapping of power spectra distribution.METHODS: Ketamine-sedated Sprague-Dawley rats were implanted to monitor behavioral states with frontoparietal electrodes in a first series of experiments and with multiple electrodes along the antero-posterior axis (F1, F2, F7, F8, T3, T4, P3, P4) in a second series. Monopolar and bipolar recordings were obtained in animals stereotaxically injected with ibotenic acid into both rostral poles of the thalamic reticular nucleus. Long-term video-EEG recordings and brain mapping based on quantitative spectral analysis were made.RESULTS: Two forms of dysrhythmia gradually emerged in the neocortical EEG at 12-24h post-injection: potentiation of theta waves and spontaneous high-voltage spindles (HVS) at 4.5-8Hz frequency. Brain mapping during these dysrhythmia shows highest power posteriorly (parietotemporal) for theta and mesiofrontally for HVS.CONCLUSIONS: Given the lack of inhibitory intrinsic interneurons in the rat thalamus, bilateral destruction of a small part of the solely GABAergic population may promote cortical dysrhythmia (probably by dis-inhibition). The topographic differences in power might indicate different involved structures.     

Epstein-Barr virus latent membrane protein 1 promotes concentration in multivesicular bodies of fibroblast growth factor 2 and its release through exosomes

FGF-2, a potent angiogenic factor that is involved in tumor invasion, is known to be released extracellularly by a nonclassical secretory pathway. Recently it has become clear that Epstein-Barr virus, specifically its oncoprotein LMP1, can induce expression of angiogenic factors. Among these factors is FGF-2. LMP1 not only promotes expression of FGF-2, but also the release extracellularly of its 18-kDa isoform. We analyzed the mechanism of FGF-2 release induced by LMP1. Confocal immunofluorescence microscopy revealed colocalization of FGF-2 with LMP1 in small dots also stained positively for CD63 and cathepsin D, markers of late endosomes or multivesicular bodies. Biochemical analysis and immunoelectron microscopy of purified exosomal fractions from cotransfected cells demonstrated increased release of exosomes and the concentration of LMP1 and FGF-2 in these structures. Moreover, cotransfection appeared to induce partial redistribution of the Na(+)/K(+)-ATPase, which participates in FGF-2 release, from the plasma membrane to the intracellular LMP1/FGF-2 positive dots. Treatment with ouabain, which inhibits Na(+)/K(+)-ATPase activity, partially suppressed FGF-2 secretion via exosomes in a dose-dependent manner. The results suggest that exosomes may represent a previously unrecognized mechanism for FGF-2 release mediated by LMP1, and that this pathway involves the activity of Na(+)/K(+)-ATPase.     

Italian version of ASES questionnaire for shoulder assessment: cross-cultural adaptation and validation

The patient-oriented measures, represented by self-administered questionnaire, have become an important aspect of clinical outcome assessment. To be used with different language groups and in different countries, questionnaires must be translated and adapted to new cultural characteristics and then validated by a widely accepted process to evaluate reliability and validity, fundamental characteristic for each measure. The aim of the study is to perform the cross-cultural adaptation and to assess the Italian version instrument reliability and validity. The study design is a cross-cultural adaptation and cross-sectional study of a sample of patients affected by shoulder disorder with a subsample followed prospectively for retest reliability. The American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form was culturally adapted for Italian-speaking people, following the simplified Guillemin criteria. Reliability and validity were assessed in a cross-sectional study of 50 consecutive patients affected by shoulder disorder. A sub-sample of 20 patients was followed prospectively for retest reliability. The results were compared with other validated patient-oriented measures. The ASES scales showed a high correlation with other patient-oriented measures, as hypothesized, and it also showed good values with regard to reproducibility, consistency and validity, to the original versions published in English. These findings suggest that the evaluation capacities of the Italian version of ASES are equivalent to those of English language version.     

Functional derangement of liver mitochondria from hexachlorobenzene-treated rats

This study investigated impairment of oxidative phosphorylation in mitochondria isolated from the liver of hexachlorobenzene (HCB)-treated rats. Partial and reversible uncoupling of the phosphorylative process was found in liver mitochondria from rats dosed with HCB for 60 days. Pentachlorophenol, endogenously formed by HCB metabolism, was detected in the mitochondria at a concentration of 0.3-0.4 nmol/mg protein. Based on the effect of pentachlorophenol, added in vitro at a concentration similar to that found in vivo, it was concluded that the uncoupling of oxidative phosphorylation under these experimental conditions was almost completely due to the presence of pentachlorophenol.     

The maturation potential of NK cell clones toward autologous dendritic cells correlates with HMGB1 secretion

Interaction of NK cells with autologous immature dendritic cells (iDCs) results in reciprocal activation. We have previously reported that NK cells trigger iDC to polarize and secrete IL-18; in turn, DC-activated NK cells secrete the nuclear protein/proinflammatory cytokine high mobility group box protein 1 (HMGB1), which induces DC maturation and prevents DC from lysis. However, activated NK cells can also kill iDC. To investigate whether effector and maturative properties may coexist or segregate in different NK subsets, human NK cell clones were generated and analyzed for their effects on iDC. We found that the ability of different NK cell clones to induce iDC maturation is unlinked to their phenotypic and cytolytic features but correlates with the relocation of HMGB1 from nucleus to cytoplasm. "Maturative" NK cell clones secrete HMGB1 spontaneously. It is interesting that secretion is strongly enhanced by engagement of the surface molecule NKp30 but only slightly induced by triggering of the activating NK receptor CD16. However, culturing freshly isolated NK cells for 1 week with low doses of anti-CD16 triggers the relocation of HMGB1 from nucleus to cytoplasm and its spontaneous secretion, resulting in a stronger maturation potential of the NK cells. Together, our data indicate that NK cells comprise functionally different subsets, endowed with different capacities to secrete HMGB1 and to induce maturation of autologous iDC. Nonetheless, maturation properties can be modulated by different stimuli. This suggests that depending on the environmental stimuli, NK/iDC interaction can lead to different outcomes, thus influencing immune response.