Glycogen storage disease type III: A novel Agl knockout mouse model

AbstractGlycogen storage disease type III is an autosomal recessive disease characterized by a deficiency in the glycogen debranching enzyme, encoded by AGL. Essential features of this disease are hepatomegaly, hypoglycemia, hyperlipidemia, and growth retardation. Progressive skeletal myopathy, neuropathy, and/or cardiomyopathy become prominent in adults. Currently, there is no available cure. We generated an Agl knockout mouse model by deletion of the carboxy terminus of the protein, including the carboxy end of the glucosidase domain and the glycogen-binding domain. Agl knockout mice presented serious hepatomegaly, but we did not observe signs of cirrhosis or adenomas. In affected tissues, glycogen storage was higher than in wild-type mice, even in the central nervous system which has never been tested in GSDIII patients. The biochemical findings were in accordance with histological data, which clearly documented tissue impairment due to glycogen accumulation. Indeed, electron microscopy revealed the disruption of contractile units due to glycogen infiltrations. Furthermore, adult Agl knockout animals appeared less prompt to move, and they exhibited kyphosis. Three-mo-old Agl knockout mice could not run, and adult mice showed exercise intolerance. In addition, older affected animals exhibited an accelerated respiratory rate even at basal conditions. This observation was correlated with severe glycogen accumulation in the diaphragm. Diffuse glycogen deposition was observed in the tongues of affected mice. Our results demonstrate that this Agl knockout mouse is a reliable model for human glycogenosis type III, as it recapitulates the essential phenotypic features of the disease

Identifying Critical Nutrient Intake in Groups at Risk of Poverty in Europe: The CHANCE Project Approach

The aim of the CHANCE project is to develop novel and affordable nutritious foods to optimize the diet and reduce the risk of diet-related diseases among groups at risk of poverty (ROP). This paper describes the methodology used in the two initial steps to accomplish the project's objective as follows: 1. a literature review of existing data and 2. an identification of ROP groups with which to design and perform the CHANCE nutritional survey, which will supply new data that is useful for formulating the new CHANCE food. Based on the literature review, a low intake of fruit and vegetables, whole grain products, fish, energy, fiber, vitamins B1, B2, B3, B6, B12 and C, folate, calcium, magnesium, iron, potassium and zinc and a high intake of starchy foods, processed meat and sodium were apparent. However, the available data appeared fragmented because of the different methodologies used in the studies. A more global vision of the main nutritional problems that are present among low-income people in Europe is needed, and the first step to achieve this goal is the use of common criteria to define the risk of poverty. The scoring system described here represents novel criteria for defining at-risk-of-poverty groups not only in the CHANCE-participating countries but also all over Europe

Effect of Sprouting on Biomolecular and Antioxidant Features of Common Buckwheat (Fagopyrum esculentum)

Buckwheat is a pseudo-cereal widely grown and consumed throughout the world. Buckwheat is recognized as a good source of nutrients and, in combination with other health-promoting components, is receiving increasing attention as a potential functional food. Despite the high nutritional value of buckwheat, a variety of anti-nutritional features makes it difficult to exploit its full potential. In this framework, sprouting (or germination) may represent a process capable of improving the macromolecular profile, including reducing anti-nutritional factors and/or synthesizing or releasing bioactives. This study addressed changes in the biomolecular profile and composition of buckwheat that was sprouted for 48 and 72 h. Sprouting increased the content of peptides and free-phenolic compounds and the antioxidant activity, caused a marked decline in the concentration of several anti-nutritional components, and affected the metabolomic profile with an overall improvement in the nutritional characteristics. These results further confirm sprouting as a process suitable for improving the compositional traits of cereals and pseudo-cereals, and are further steps towards the exploitation of sprouted buckwheat as a high-quality ingredient in innovative products of industrial interest

Evaluation of human gene variant detection in amplicon pools by the GS-FLX parallel Pyrosequencer

<p>Abstract</p> <p>Background</p> <p>A new priority in genome research is large-scale resequencing of genes to understand the molecular basis of hereditary disease and cancer. We assessed the ability of massively parallel pyrosequencing to identify sequence variants in pools. From a large collection of human PCR samples we selected 343 PCR products belonging to 16 disease genes and including a large spectrum of sequence variations previously identified by Sanger sequencing. The sequence variants included SNPs and small deletions and insertions (up to 44 bp), in homozygous or heterozygous state.</p> <p>Results</p> <p>The DNA was combined in 4 pools containing from 27 to 164 amplicons and from 8,9 to 50,8 Kb to sequence for a total of 110 Kb. Pyrosequencing generated over 80 million base pairs of data. Blind searching for sequence variations with a specifically designed bioinformatics procedure identified 465 putative sequence variants, including 412 true variants, 53 false positives (in or adjacent to homopolymeric tracts), no false negatives. All known variants in positions covered with at least 30× depth were correctly recognized.</p> <p>Conclusion</p> <p>Massively parallel pyrosequencing may be used to simplify and speed the search for DNA variations in PCR products. Our results encourage further studies to evaluate molecular diagnostics applications.</p

Long Term Assessment of Anti-SARS-CoV-2 Immunogenicity after mRNA Vaccine in Persons Living with HIV

(1) Background: Waning of neutralizing and cell-mediated immune response after the primary vaccine cycle (PVC) and the first booster dose (BD) is of concern, especially for PLWH with a CD4 count ≤200 cells/mm3. (2) Methods: Neutralizing antibodies (nAbs) titers by microneutralization assay against WD614G/Omicron BA.1 and IFNγ production by ELISA assay were measured in samples of PLWH at four time points [2 and 4 months post-PVC (T1 and T2), 2 weeks and 5 months after the BD (T3 and T4)]. Participants were stratified by CD4 count after PVC (LCD4, ≤200/mm3; ICD4, 201–500/mm3, and HCD4, &gt;500/mm3). Mixed models were used to compare mean responses over T1–T4 across CD4 groups. (3) Results: 314 PLWH on ART (LCD4 = 56; ICD4 = 120; HCD4 = 138) were enrolled. At T2, levels of nAbs were significantly lower in LCD4 vs. ICD4/HCD4 (p = 0.04). The BD was crucial for increasing nAbs titers above 1:40 at T3 and up to T4 for WD614G. A positive T cell response after PVC was observed in all groups, regardless of CD4 (p = 0.31). (4) Conclusions: Waning of nAbs after PVC was more important in LCD4 group. The BD managed to re-establish higher levels of nAbs against WD614G, which were retained for 5 months, but for shorter time for Omicron BA.1. The T cellular response in the LCD4 group was lower than that seen in participants with higher CD4 count, but, importantly, it remained above detectable levels over the entire study period

Local site effects estimation at Amatrice (Central Italy) through seismological methods

We present the results of seismological and geophysical investigations performed by the “Istituto Nazionale di Geofisica e Vulcanologia” team operating in Amatrice village (Central Italy), in the emergency phases following the Mw 6.0 event of August 24th 2016, that caused severe damage in downtown and surrounding areas. Data from seven seismic stations equipped with both weak and strong motion sensors are analyzed in terms of standard spectral ratio to empirically define amplification function using a bedrock reference site. Ambient vibration spectral ratios between horizontal and vertical component of motion are also evaluated in a large number of sites, spread out in the investigated area, to recover the resonance frequency of the soft soil outcropping layers and to generalize the results obtained by earthquake data. Ambient noise vibration are also used for applying a 2D array approach based on surface waves techniques in order to define the near-surface velocity model and to verify its lateral variation. The results allows to better understand the amplification factors in the investigated area, showing spatial variation of site effects despite of the homogeneous shallow geological condition indicated by the microzonation studies available at moment of the described field campaign. The analysis reveals a diffuse amplification effect which reaches its maximum values in downtown area with a resonant frequency of about 2 Hz. The obtained results were used to integrate the microzonation studies and they can be used for urban planning and reconstruction activities.Published5713–57394T. Sismicità dell'ItaliaJCR Journa

Site effects estimation and their effects on strong ground motion at Amatrice village (Central Italy)

We present a summary of seismological and geophysical investigations at Amatrice (Central Italy), a village seated on an alluvial terrace and severely stroke by the Mw 6.0 event of August 24th 2016. The high vulnerability alone could not explain the heavy damage (X-XI MCS), whereas the vicinity of the seismic source and the peculiar site effects should be claimed to understand the ground motion variability. After the first mainshock, we investigated the Amatrice terrace for microzonation purposes together with several Italian institutions (Priolo et al., Bull. Earthquake Eng. 2019). In particular: (i) we installed 7 seismic stations as a part of the 3A network (DOI: 10.13127/SD/ku7Xm12Yy9; Cara et al., Sci. Data 2019); we performed (ii) an extensive campaign of 60 single-station ambient noise measurements (downtown stations recorded also few earthquakes), and (iii) several 2D passive seismic arrays aimed at obtaining Vs profiles down to a depth of few tens of meters (Milana et al., Bull. Earthquake Eng. 2019). Earthquake recordings were used to empirically evaluate ground motion amplification effects through spectral ratio approaches, and noise data were collected for defining the spatial distribution of the resonance frequencies. Data analysis reveals a diffuse amplification effect that reaches its maximum values in downtown area with a resonant frequency (f0) of about 2 Hz. Seismic amplification is also characterized by spatial variation and directional amplification, mainly in downtown to the west side of the alluvial terrace, and related to both stratigraphic and topographic effects. This effect tends to decrease and almost vanishes in the central part of the terrace, and it increases again moving towards its eastern edge with a clear shift of f0 towards higher frequencies. Empirical transfer functions were then used to recover the ground motion that could have hit the historical center of Amatrice during the August 24th mainshock, through the convolution with the only record in the vicinity (IT.AMT station experienced a PGA of 0.87 g). The reconstructed peak values are much greater than expected from ground motion models, showing that detailed studies on local site response can largely modify the seismic hazard assessment.PublishedSan Francisco, California (USA)4T. Sismicità dell'Italia5T. Sismologia, geofisica e geologia per l'ingegneria sismic

Local variability of the ground shaking during the 2009 L’Aquila earthquake (April 6, 2009—Mw 6.3): the case study of Onna and Monticchio villages

The 2009 Mw 6.3 L’Aquila event caused extensive damage in the city of L’Aquila and in some small towns in its vicinity. The most severe damage was recognized SE of L’Aquila town along the Aterno river valley. Although building vulnerability and near-source effects are strongly responsible for the high level of destruction, site effects have been invoked to explain the damage heterogeneities and the similarities between the 2009 macroseismic field with the intensities of historical earthquakes. The small village of Onna is settled on quaternary alluvium and suffered during the L’Aquila event an extremely heavy damage in the masonry structures with intensity IX–X on the Mercalli-Cancani-Sieberg (MCS) scale. The village of Monticchio, far less than 1.3 km from Onna, is mostly situated on Meso- zoic limestone and suffered a smaller level of damaging (VI MCS). In the present paper, we analyze the aftershock recordings at seismic stations deployed in a small area of the middle-Aterno valley including Onna and Monticchio. The aim is to investigate local ampli-fication effects caused by the near-surface geology. Because the seismological stations are close together, vulnerability and near-source effects are assumed to be constant. The wave- form analysis shows that the ground motion at Onna is systematically characterized by large high-frequency content. The frequency resonance is varying from 2 to 3 Hz and it is related to alluvial sediments with a thickness of about 40 m that overlay a stiffer Pleistocene substrate. The ground motion recordings of Onna are well reproduced by the predictive equation for the Italian territory.Published783-8072T. Sorgente SismicaJCR Journalreserve

PMN-MDSC frequency discriminates active versus latent tuberculosis and could play a role in counteracting the immune-mediated lung damage in active disease

: Tuberculosis (TB), due to Mycobacterium tuberculosis infection, is still the principal cause of death caused by a single infectious agent. The balance between the bacillus and host defense mechanisms reflects the different manifestations of the pathology. Factors defining this variety are unclear and likely involve both mycobacterial and immunological components. Myeloid derived suppressor cells (MDSC) have been shown to be expanded during TB, but their role in human TB pathogenesis is not clear. We evaluated the frequency of circulating MDSC by flow-cytometry in 19 patients with active TB, 18 with latent TB infection (LTBI), and 12 healthy donors (HD) as control. Moreover, we investigated the capacity of MDSC to modulate the mycobactericidal activity of monocytes. The association between MDSC level and TB chest X-ray severity score was analyzed. We observed that, unlike active TB, polymorphonuclear (PMN)-MDSC are not expanded in LTBI patients, and, by performing a receiver operating characteristic (ROC) curve analysis, we found that PMN-MDSC frequency supported the discrimination between active disease and LTBI. Interestingly, we observed an association between PMN-MDSC levels and the severity of TB disease evaluated by chest X-ray. Specifically, PMN-MDSC frequency was higher in those classified with a low/mild severity score compared to those classified with a high severity score. Moreover, PMN-MDSC can impact mycobacterial growth by inducing ROS production in Bacillus Calmette et Guerin (BCG)-infected monocytes. This effect was lost when tested with M. tuberculosis (MTB), In conclusion, our data indicate that the elevated frequency of PMN-MDSC in IGRA-positive individuals is associated with active TB. Our findings also pointed out a beneficial role of PMN-MDSC during human active TB, most likely associated with the limitation of inflammation-induced tissue damage

Site effect studies following the 2016 Mw 6.0 Amatrice Earthquake (Italy): the Emersito Task Force activities

On August 24, 2016, at 01:36 UTC a MW 6.0 earthquake struck an extensive area of the Central Apennines (Italy) be-tween the towns of Norcia and Amatrice. Due to the mainshock magnitude and the widespread damaging level of build-ings in the epicentral area, the Emersito task force has been mobilized by the Istituto Nazionale di Geofisica e Vulcanologia (INGV). The aim of Emersito is to carry out and coordinate the monitoring of local site effects, caused by geological and geomorphological settings. During the first days of the seismic emergency, Emersito installed a tempo-rary seismic network for site effect studies at 4 municipalities close to the epicentral area (Amandola, Civitella del Tronto, Montereale and Capitignano), using 22 stations equipped with both velocimetric and accelerometric sensors. The selection of the sites where stations have been installed was mainly driven by the proximity to the epicentral area (without interfere with the rescue operations) and by peculiar geologic and geomorphologic settings (topographic irregu-larities, fault zones, alluvial plains). Preliminary analyses performed on ambient noise and aftershocks signals show that directional amplification effects may have occurred at stations installed on the top of topographic irregularities. We also observed the lengthening and amplification of the seismograms and a variability of the peaked frequency across the sedi-mentary basin between Montereale and Capitignano, probably related to a different thickness of the deposits. Further analyses are necessary to assess the correlation with surface geology.Published4T. Sismologia, geofisica e geologia per l'ingegneria sismica1SR. TERREMOTI - Servizi e ricerca per la Società1IT. Reti di monitoraggioJCR Journa