Detection of Trypanosoma cruzi and Trypanosoma rangeli infection by duplex PCR assay based on telomeric sequences

We used the species specificity and repetitious nature of subtelomeric kinetoplastida sequences to generate a duplex PCR assay for the simultaneous detection of Trypanosoma cruzi and Trypanosoma rangeli in experimentally and naturally infected triatomine (Reduviid) bugs and in infected human subjects. The assay was species specific and was capable of detecting 1/20th of T. cruzi and 1/4th of T. rangeli cell equivalents without complementary hybridization. In addition, the PCR-based assay was robust enough for direct application to difficult biological samples such as Reduviid feces or guts and was capable of recognizing all T. cruzi and T. rangeli strains and lineages. Because the assay primers amplify entirely different target sequences, no reaction interference was observed, facilitating future adaptation of this assay to an automated format.     

Development of the anatomical alteration of the cerebellar fissura prima

The development of the naturally occurring malformation of the cerebellar fissura prima was monitored in rats starting from 4 days of life to the adulthood. The first sign of the malformation was evident at 10 days of life and consisted of an interruption of the pia mater and the fusion of the external granular layers on the two sides of the fissura. Later, nests of apparently mature granule cells could be seen to be encircled by cells of the external granular layer and to be connected to the granule cell layer by thin bridges of cells. Calretinin immunoreactive fibers followed the bridges of cells to reach the ectopic masses of cells. Towards the end of histogenesis and in adult animals, brush cells and Golgi cells were present in the ectopic masses of granule cells. The latter appeared to contribute to the formation of normal glomeruli, as in the orthotopic granule cell layer. In addition, bundles of parallel fibers crossed the boundary between the molecular layers on the two side of the fissure, thus suggesting that parallel fibers can contact Purkinje cells of the opposite folium.     

Damage and repair of the immature rat cerebellum after cis-dichlorodiammineplatinum II (cis-DDP) treatment. An ultrastructural study

Summary The aim of this electron microscopy study was to further investigate the effects of cis-dichlorodiammineplatinum (cis-DDP) on the cerebellum of the immature rat. Ten-day-old animals were treated with cis-DDP subcutaneously and killed after 1, 7, 15 or 21 days. On postinjection day 1, cis-DDP effects were evident mainly in the external granular layer, with nuclear damage in many dividing cells, while their cytoplasm appeared to be less affected. Some binucleate cells were also present. On the contrary, in postmitotic or more differentiated cells, only cytoplasmic alterations were found. At later stages (postinjection day 7), the frequency of damaged cells in the external granular layer decreased, but there was a cellular deficit in the internal granular layer. Many postmitotic neurons underwent coagulative necrosis. Finally (postinjection days 15 and 21), the cellular deficit was partly compensated for by reactive structures, e.g., glial cell fibers, which underwent hypertrophy after initial edema. Moreover, packing densities of Bergmann astrocytes and oligodendrocytes were higher.This study is a part of the scientific exchange program Proliferation and differentiation of normal and tumor cells between the Italian National Research Council and the Czechoslovak Academy of Sciences     

Neuroprotective effects of α-dihydroergocryptine against damages in the substantia nigra caused by severe treatment with 1-methyl-4-phenyl-1,3,3,6-tetrahydropyridine

Summary The effects on the substantia nigra of -dihydroergocryptine (DEK), a drug with strong dopaminomimetic activity, were tested with a severe 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment in monkeys. Compared with monkeys treated with MPTP alone, the animals which received DEK plus MPTP showed reduced neuronal death in the substantia nigra. The reactive astrocytes were increased in number. Moreover, several axons which were immunopositive to phosphorylated neurofilament proteins and with features similar to those of control animals were seen in the pars compacta. The findings suggest that DEK preserves neuronal morphology and brain architecture.supported by Italian M.U.R.S.T. grants (to G.B.) and by Poli S. P. A., Rozzano, Milano     

Internet Addiction Test (IAT): Which is the Best Factorial Solution?

Background

The Internet Addiction Test (IAT) by Kimberly Young is one of the most utilized diagnostic instruments for Internet addiction. Although many studies have documented psychometric properties of the IAT, consensus on the optimal overall structure of the instrument has yet to emerge since previous analyses yielded markedly different factor analytic results.

Objective

The objective of this study was to evaluate the psychometric properties of the Italian version of the IAT, specifically testing the factor structure stability across cultures.

Methods

In order to determine the dimensional structure underlying the questionnaire, both exploratory and confirmatory factor analyses were performed. The reliability of the questionnaire was computed by the Cronbach alpha coefficient.

Results

Data analyses were conducted on a sample of 485 college students (32.3%, 157/485 males and 67.7%, 328/485 females) with a mean age of 24.05 years (SD 7.3, range 17-47). Results showed 176/485 (36.3%) participants with IAT score from 40 to 69, revealing excessive Internet use, and 11/485 (1.9%) participants with IAT score from 70 to 100, suggesting significant problems because of Internet use. The IAT Italian version showed good psychometric properties, in terms of internal consistency and factorial validity. Alpha values were satisfactory for both the one-factor solution (Cronbach alpha=.91), and the two-factor solution (Cronbach alpha=.88 and Cronbach alpha=.79). The one-factor solution comprised 20 items, explaining 36.18% of the variance. The two-factor solution, accounting for 42.15% of the variance, showed 11 items loading on Factor 1 (Emotional and Cognitive Preoccupation with the Internet) and 7 items on Factor 2 (Loss of Control and Interference with Daily Life). Goodness-of-fit indexes (NNFI: Non-Normed Fit Index; CFI: Comparative Fit Index; RMSEA: Root Mean Square Error of Approximation; SRMR: Standardized Root Mean Square Residual) from confirmatory factor analyses conducted on a random half subsample of participants (n=243) were satisfactory in both factorial solutions: two-factor model (χ² ₁₃₂= 354.17, P<.001, χ²/df=2.68, NNFI=.99, CFI=.99, RMSEA=.02 [90% CI 0.000-0.038], and SRMR=.07), and one-factor model (χ² ₁₆₉=483.79, P<.001, χ²/df=2.86, NNFI=.98, CFI=.99, RMSEA=.02 [90% CI 0.000-0.039], and SRMR=.07).

Conclusions

Our study was aimed at determining the most parsimonious and veridical representation of the structure of Internet addiction as measured by the IAT. Based on our findings, support was provided for both single and two-factor models, with slightly strong support for the bidimensionality of the instrument. Given the inconsistency of the factor analytic literature of the IAT, researchers should exercise caution when using the instrument, dividing the scale into factors or subscales. Additional research examining the cross-cultural stability of factor solutions is still needed.     

In vitro evaluation of permeation,toxicity and effect of praziquantel-loaded solid lipid nanoparticles against Schistosoma mansoni as a strategy to improve efficacy of the schistosomiasis treatment

Solid lipid nanoparticles (SLN) are a promising drug delivery system for oral administration of poorly-water soluble drugs because of their capacity to increase the solubility of drug molecules when loaded in their lipid matrices, with the resulting improvement of the drug bioavailability. In the present work, we have developed praziquantel (PZQ)-loaded SLN and explored the biological applications of this system for intestinal permeation of PZQ. The effect in vitro on Schistosoma mansoni culture and the cytotoxicity in HepG2 line cell were also evaluated. The results showed a significant decrease in the intestinal absorption of PZQ loaded in SLN compared to free PZQ, suggesting that the SLN matrix could act as reservoir system. In culture of S. mansoni, we observed that PZQ-loaded SLN were more effective than free PZQ, leading the death of the parasites in less time. The result was proportional to doses of PZQ (25 and 50 μg mL⁻¹) and lipid concentration. Regarding cytotoxicity, the encapsulation of PZQ into SLN decreased the toxicity in HepG2 cells in comparison to the free PZQ. From the obtained results, PZQ-loaded SLN could be a new drug delivery system for the schistosomiasis treatment especially in marginalized communities, improving the therapeutic efficacy and reducing the toxic effects of PZQ.     

The non-alcoholic fraction of beer increases stromal cell derived factor 1 and the number of circulating endothelial progenitor cells in high cardiovascular risk subjects: A randomized clinical trial

Rationale

Moderate alcohol consumption is associated with a decrease in cardiovascular risk, but fermented beverages seem to confer greater cardiovascular protection due to their polyphenolic content. Circulating endothelial progenitor cells (EPC) are bone-marrow-derived stem cells with the ability to repair and maintain endothelial integrity and function and are considered as a surrogate marker of vascular function and cumulative cardiovascular risk. Nevertheless, no study has been carried out on the effects of moderate beer consumption on the number of circulating EPC in high cardiovascular risk patients.

Objective

To compare the effects of moderate consumption of beer, non-alcoholic beer and gin on the number of circulating EPC and EPC-mobilizing factors.

Methods

In this crossover trial, 33 men at high cardiovascular risk were randomized to receive beer (30 g alcohol/d), the equivalent amount of polyphenols in the form of non-alcoholic beer, or gin (30 g alcohol/d) for 4 weeks. Diet and physical exercise were carefully monitored.

Results

The number of circulating EPC and EPC-mobilizing factors were determined at baseline and after each intervention. After the beer and non-alcoholic beer interventions, the number of circulating EPC significantly increased by 8 and 5 units, respectively, while no significant differences were observed after the gin period. In correlation, stromal cell derived factor 1 increased significantly after the non-alcoholic and the beer interventions.

Conclusions

The non-alcoholic fraction of beer increases the number of circulating EPC in peripheral blood from high cardiovascular risk subjects.

Clinical trial registration

http://www.controlled-trials.com/ISRCTN95345245 ISRCTN95345245     

Information and communication technology in chronic diseases: a patient’s opportunity

The increasing aging population, the prevalence of chronic diseases and rising costs have brought about some unique health care challenge to our global society. In response to the unmet health care needs, researchers are actively seeking for innovative solutions that target for (1) prevention of diseases and (2) personalized diagnosis and treatment. It is envisaged that by taking preventive measures for health monitoring, diagnosing and treating patients with a personalized approach at an early stage of disease development, health care will be more cost effective and sustainable. The authors provide an overview of the advancements in Information and Communication Technology (ICT), and explain how some innovative health solutions, through the use of Telemedicine, can now be an opportunity for patients and their family.     

Prolidase enzyme is required for extracellular matrix integrity and impacts on postnatal cerebellar cortex development

The extracellular matrix is essential for brain development, lamination, and synaptogenesis. In particular, the basement membrane below the pial meninx (pBM) is required for correct cortical development. The last step in the catabolism of the most abundant protein in pBM, collagen Type IV, requires prolidase, an exopeptidase cleaving the imidodipeptides containing pro or hyp at the C-terminal end. Mutations impairing prolidase activity lead in humans to the rare disease prolidase deficiency characterized by severe skin ulcers and mental impairment. Thus, the dark-like (dal) mouse, in which the prolidase is knocked-out, was used to investigate whether the deficiency of prolidase affects the neuronal maturation during development of a brain cortex area. Focusing on the cerebellar cortex, thinner collagen fibers and disorganized pBM were found. Aberrant cortical granule cell proliferation and migration occurred, associated to defects in brain lamination, and in particular in maturation of Purkinje neurons and formation of synaptic contacts. This study deeply elucidates a link between prolidase activity and neuronal maturation shedding new light on the molecular basis of functional aspects in the prolidase deficiency.