Rheumatoid arthritis and anti-thyroid antibodies

The aim of this study was to evaluate the quality of B cell responses in patients with Inflammatory Bowel Disease (IBD) and healthy individuals of different ages, vaccinated with the pandemic (p)2009 influenza vaccine. The in vivo response was measured by the hemagglutination inhibition (HAI) assay, which represents the most established correlate with vaccine protectiveness. The in vitro response was measured by activation-induced cytidine deaminase (AID) in cultures of vaccine-stimulated PBMC. Both responses are somewhat impaired in IBD patients undergoing anti-TNF-α treatment but these are much more decreased in IBD patients undergoing treatment with anti-TNF-α and immunosuppressive (IS) drugs. These latter patients had in vivo and in vitro B cell responses similar to those of elderly individuals. Moreover, as we have previously demonstrated in healthy subjects, the in vitro response to the polyclonal stimulus CpG may be used as a biomarker for subsequent vaccine response and AID activation is correlated with the serum response in IBD patients, as it is in healthy individuals. These results altogether indicate that IBD patients on anti-TNF-α and IS have significantly impaired in vivo and in vitro B cell responses, as compared to those on monotherapy. This is the first report to demonstrate that B cell defects, as measured by the autonomous AID reporter, in IBD patients contribute to reduced humoral responses to the influenza vaccine, as we have previously shown for elderly individuals.     

Inhibition of VTA neurons activates the centrally projecting Edinger–Westphal nucleus: Evidence of a stress–reward link?

The primary site of urocortin 1 (Ucn1) expression in the brain is the centrally projecting Edinger–Westphal nucleus. The EWcp is innervated by dopaminergic neurons of the ventral tegmental area (VTA). To investigate whether activity of EWcp is regulated by the VTA, we investigated the effects of local pharmacological inhibition of VTA activity on the induction of Fos immunoreactivity in the EWcp of male C57BL/6J mice. A unilateral intracranial administration of the GABA agonist muscimol aimed at the VTA resulted in increased number of Fos-positive cells in the EWcp. This induction was lower than that produced by an intraperitoneal injection of 2.5 g/kg of ethanol. To investigate whether inhibition of dopaminergic neurons was responsible for induction of Fos, a second experiment was performed where the dopamine agonist quinpirole was unilaterally injected targeting the VTA. Injections of quinpirole also significantly induced Fos in the EWcp neurons. The induction occurred only on the side of the EWcp ipsilateral to the VTA injection. These results indicate that activity of EWcp is inhibited by tonic activity of dopaminergic VTA neurons, and that unilateral projections of VTA onto Ucn1-containing EWcp neurons provide a link between systems regulating approach and avoidance behaviors.     

IMMUNOGENICITY AND SPECIFICITY OF THE CANDIDATE MULTI-EPITOPE-VACCINES AGAINST HIV-1

The failure of some candidate HIV-1 vaccines may result from inducing very weak neutralization activity against representative primary viral isolates. Based on our hypothesis that epitope-vaccine may be a new strategy to induce high levels of neutralizing antibodies against HIV-1, we designed two candidate multi-epitope-vaccines, EP1 [C-G-(ELDKWA-GPGRAFY)₂-K] and EP2 (CG-GPGRAFY-G-ELDKWA-G-RILAVERYLKD), containing three neutralizing epitopes (GPGRAFY, ELDKWA and RILAVERYLKD) on HIV-1 envelope protein, and expected them to induce epitope-specific antibodies of predefined epitope-specificity. The two peptides were conjugated to carrier protein bovine serum albumin (BSA) and used for immunization of rabbits. Proteins were purified from the rabbit sera induced by both candidate multi-epitope-vaccines (EP1-BSA and EP2-BSA) through affinity chromatography with epitope-peptide-conjugated sepharose-column, and identified as antibodies in silver-staining and immunoblotting. These antibodies were demonstrated to recognize three neutralizing epitopes on peptides and the recombinant gp41 in ELISA-assay and immunoblotting. These results indicated that both candidate multi-epitope-vaccines could induce high levels of antibodies of predefined epitope-specificity which recognized a few of neutralizing epitopes on peptides and protein, providing experimental evidence for the new strategy to develop an effective neutralizing-antibody-based multi-epitope-vaccine against HIV-1.     

Novel light-curable materials containing experimental bioactive micro-fillers remineralise mineral-depleted bonded-dentine interfaces

This study aimed at evaluating the therapeutic remineralising effects of innovative light-curable materials (LCMs) containing two experimental calcium silicate-based micro-fillers (TCS) modified with β-TCP only or β-TCP, zinc oxide (ZnO)/polyacrylic acid (PAA) on mineral-depleted bonded-dentine interfaces in simulated body fluids (SBFS). Three experimental LCMs were formulated: (1) resin A, containing a β-TCP-modified TCS (βTCS) micro-filler; (2) resin B, containing a polycarboxylated β-TCP/ZnO-modified TCS (βZn-TCS) micro-filler; and 3) resin C, containing no filler (control). Acid-etched (35% H₃PO₄) dentine specimens were bonded using the three LCMs and submitted to atomic force microscope (AFM)/nano-indentation analysis to evaluate the modulus of elasticity (Ei) and hardness (Hi) across the interface after SBFS storage (24?h/1?m/3?m). The ultramorphology and micropermeability of the resin–dentine interface were evaluated using confocal laser microscopy. Resin–dentine sticks were created and submitted to microtensile bond strength (μTBS) test (SBFS: 24?h/3?m). Scanning electron microscopy (SEM) was performed after de-bonding. The LCMs containing the experimental bioactive micro-fillers reduced the micropermeability and induced a significant increase of the Ei and Hi along the bonding interface. The specimens created using the resin B (βZn-TCS) attained the highest μTBS values both after 24?h and 3?m of SBFS storage. In conclusion, the innovative bioactive light-curable materials tested in this study are able to induce a therapeutic remineralising effect on the nano-mechanical properties and on the sealing ability of mineral-depleted resin–dentine interfaces. The contemporary idea of minimally invasive operative treatment, where therapeutic restorations are performed to combat the carious process and remineralise the dental hard tissues, may be satisfied by using such resin-base systems, containing βTCS or βZn-TCS bioactive micro-fillers.     

Specific interpretation of hair concentrations in 2 fatal metformin intoxication cases

Hair analysis is very useful for toxicological investigations since, by providing a wider detection window, it gives the possibility to perform a retrospective study on the historical consumption of a substance. Unfortunately, there are no data available for hair concentrations in metformin-related deaths. In this study, the authors present 2 cases of fatal metformin intoxication in which, for the first time, hair analysis was performed using a specific GC–MS/MS method. Metformin was tested positive in femoral blood (112.3 mg/L and 64.7 mg/L respectively) and cardiac blood (226.9 and 203.2 mg/L) of the two subjects. For case 1, other samples were also tested positive, including vitreous humor (31.1 mg/L) and gastric contents (773.5 mg/L). In case 2, metformin was measured at 844.9 mg/L in urine. Metformin hair concentrations were 28.3–44.8 and 22.5 ng/mg for both cases, respectively. The concentrations found in the 2 fatal cases are clearly higher than those obtained in a previous study with subjects under treatment (0.3–3.8 ng/mg) or those found in 3 post-mortem cases where metformin death was excluded (0.6–1.4 ng/mg). Excessive sweating during the agonal phase due to fatal hypoglycemia could explain these elevated concentrations as sweat can have contaminated the hair.     

Body size and growth: The significance of chronic malnutrition among the Casiguran Agta

Primary objective: The Agta are a short statured, hunter–gatherer population who have been living under demographic and environmental stress. This article shows the influence of these factors and the resulting nutritional deficiency on the anthropometric measurements and mortality of Agta individuals.

Research design: The joint analysis of cross-sectional measurements of Agta children and adults and of the mortality schedules in this population aims to stress the influence of environmental pressures on the ongoing evolution of short stature.

Methods and procedures: Recumbent length, height, weight, mid-upper arm circumference, and triceps skinfolds were taken using standard methods from a total of 253 Agta individuals. Exact or nearly exact ages were taken from a long-term demographic database. z-Scores and growth curves were calculated using the NutStat program and 2000 CDC reference. Mortality schedules are from published material, improved through interviews in the field.

Main outcomes and results: Agta individuals are both short and thin when compared to other populations. Thirty-four per cent of the adults are under-nourished, while 17% of the children are wasted, according to international standards. A major and delayed peak of mortality in early infancy overlaps with a period of average decrease in body length in relation to the reference.

Conclusion: Demographic indicators of poor health-related quality of life are consistent with slowed patterns of growth observed, stressing the importance of environmental pressure in maintaining the short stature of the Agta population.