Magnetic hyperthermia and oxidative damage to dna of human hepatocarcinoma cells

Nanotechnology is addressing major urgent needs for cancer treatment. We conducted a study to compare the frequency of 3-(2-deoxy-β-d-erythro-pentafuranosyl)pyrimido[1,2-α]purin-10(3H)-one deoxyguanosine (M1dG) and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) adducts, biomarkers of oxidative stress and/or lipid peroxidation, on human hepatocarcinoma HepG2 cells exposed to increasing levels of Fe3O4-nanoparticles (NPs) versus untreated cells at different lengths of incubations, and in the presence of increasing exposures to an alternating magnetic field (AMF) of 186 kHz using 32P-postlabeling. The levels of oxidative damage tended to increase significantly after ≥24 h of incubations compared to controls. The oxidative DNA damage tended to reach a steady-state after treatment with 60 μg/mL of Fe3O4-NPs. Significant dose–response relationships were observed. A greater adduct production was observed after magnetic hyperthermia, with the highest amounts of oxidative lesions after 40 min exposure to AMF. The effects of magnetic hyperthermia were significantly increased with exposure and incubation times. Most important, the levels of oxidative lesions in AMF exposed NP treated cells were up to 20-fold greater relative to those observed in nonexposed NP treated cells. Generation of oxidative lesions may be a mechanism by which magnetic hyperthermia induces cancer cell death

Fatigue as hallmark of Fabry disease: role of bioenergetic alterations

Fabry disease (FD) is a lysosomal storage disorder due to the impaired activity of the α-galactosidase A (GLA) enzyme which induces Gb3 deposition and multiorgan dysfunction. Exercise intolerance and fatigue are frequent and early findings in FD patients, representing a self-standing clinical phenotype with a significant impact on the patient's quality of life. Several determinants can trigger fatigability in Fabry patients, including psychological factors, cardiopulmonary dysfunctions, and primary alterations of skeletal muscle. The “metabolic hypothesis” to explain skeletal muscle symptoms and fatigability in Fabry patients is growing acknowledged. In this report, we will focus on the primary alterations of the motor system emphasizing the role of skeletal muscle metabolic disarrangement in determining the altered exercise tolerance in Fabry patients. We will discuss the most recent findings about the metabolic profile associated with Fabry disease offering new insights for diagnosis, management, and therapy

Women in Zimbabwe: the Last Fifteen Years

This study examines the roles and status of women in Zimbabwe before and after Independence in 1980. It focuses on whether or not women actually have rights now that Independence has occurred. The review of literature examines several research studies all discussing before, during and after Independence in Zimbabwe. The study also discusses several theories including, functionalism, conflict, elite and eclectic feminist theories. The methods used in this study were personal interviews and questionnaires in the mail. The results section includes both FREQUECIES and T-TEST runs. In sum, the paper is a view of the status of black women in Zimbabwe before, during and, especially, after Independence. Results showed that black Zimbabwean women do enjoy a greater economic, political and social power than they enjoyed before Independence. Future research will be undertaken to further this study over a longer period of time in order to obtain a larger sample

Analisi di sbilanciamenti sul cromosoma 1q21.1-q21.2 individuati con il array-CGH

Molti casi di disabilità intellettive, ritardi psicomotori e di sviluppo sono determinati da un’alterazione genomica, che spesso rimane non identificata. Comprendere la causa di un fenotipo patologico è spesso fondamentale per la diagnosi e per la terapia. Nel corso degli anni i citogenetisti hanno avuto a disposizione strumenti di analisi sempre più sofisticati e risolutivi, passando dal cariotipo convenzionale a tecniche di citogenetica molecolare, fino ad arrivare all’array-CGH (Microarray Comparative Genomic Hybridization). Questa metodica, relativamente recente, ha consentito un notevole aumento della capacità di rilevazione di alterazioni genomiche e ha permesso l’individuazione di nuove sindromi da microdelezione e microduplicazione, evidenziando inoltre una variabilità del genoma umano finora non sospettata. Essa si basa sull’ibridazione competitiva del DNA del paziente (test) e del DNA di un campione di riferimento (reference) e consente di evidenziare sbilanciamenti genomici submicroscopici, nell’ordine delle 100 Kb. Durante il tirocinio presso il laboratorio di Genetica Medica dell’Azienda Ospedaliero - Universitaria Pisana sono stati analizzati i DNA di 150 soggetti con la tecnica dell’array-CGH. Tra i pazienti che sono risultati avere uno sbilanciamento, è stato selezionato un gruppo di 8 casi che condividono un’alterazione sul cromosoma 1 nella regione q21.1-q21.2. La regione minima comune coinvolta nello sbilanciamento è di circa 1,2 Mb, che in tre pazienti risulta deleta e in due duplicata; nei restanti casi l’alterazione è più ampia: sono state identificate due sorelle con una delezione di circa 2,8 Mb ed un caso con una duplicazione di circa 8 Mb. La regione minima deleta o duplicata contiene 11 geni. Per cercare di comprendere il meccanismo molecolare alla base di questi sbilanciamenti sono stati esaminati in dettaglio la regione 1q21.1-q21.2 e, in particolare, sono state analizzate le sequenze fiancheggianti la regione minima comune. In esse sono presenti numerose duplicazioni segmentali che probabilmente causano eventi di ricombinazione omologa non allelica (NAHR), generando duplicazioni e delezioni. Sono stati poi analizzati anche i DNA dei genitori per verificare se gli sbilanciamenti fossero de novo o ereditati; nella maggior parte dei casi l’alterazione è ereditata dal padre o dalla madre, che tuttavia non presenta un quadro patologico. Questi pazienti, come quelli con sbilanciamenti 1q21.1 riportati in letteratura, mostrano una notevole variabilità fenotipica non riconducibile ad una sindrome. Questa ampia variabilità può essere attribuita a fenomeni di penetranza incompleta, come pure a fenomeni epigenetici quali l’imprinting, la variazione nell’espressione di geni presenti nella regione coinvolta, e, nel caso di delezioni, allo smascheramento di varianti recessive presenti sull’unico allele rimasto. Le alterazioni della regione 1q21.1-q21.2 evidenziano come un’analisi tramite array-CGH possa essere fondamentale nella diagnosi di pazienti con un fenotipo non sindromico e sono quindi un chiaro esempio di come si possa giungere alla diagnosi con un percorso a ritroso, dal genotipo al fenotipo

Phenotype microarray analysis may unravel genetic determinants of the stress response by Rhodococcus aetherivorans BCP1 and Rhodococcus opacus R7

In the present study, the response of Rhodococcus aetherivorans BCP1 and Rhodococcus opacus R7 to various stress conditions and several antimicrobials was examined by PM in relation with genetic determinants, as revealed by annotation analysis of the two genomes. Comparison between metabolic activities and genetic features of BCP1 and R7 provided new insight into the environmental persistence of these two members of the genus Rhodococcus

Genome and Phenotype Microarray analyses of Rhodococcus sp. BCP1 and Rhodococcus opacus R7: genetic determinants and metabolic abilities with environmental relevance

In this paper comparative genome and phenotype microarray analyses of Rhodococcus sp. BCP1 and Rhodococcus opacus R7 were performed. Rhodococcus sp. BCP1 was selected for its ability to grow on short-chain n-alkanes and R. opacus R7 was isolated for its ability to grow on naphthalene and on o-xylene. Results of genome comparison, includ- ing BCP1, R7, along with other Rhodococcus reference strains, showed that at least 30% of the genome of each strain presented unique sequences and only 50% of the predicted proteome was shared. To associate genomic features with metabolic capabilities of BCP1 and R7 strains, hundreds of different growth conditions were tested through Phenotype Microarray, by using Biolog plates and plates manually prepared with additional xenobiotic compounds. Around one-third of the surveyed carbon sources was utilized by both strains although R7 generally showed higher metabolic activity values compared to BCP1. More- over, R7 showed broader range of nitrogen and sulphur sources. Phenotype Microarray data were combined with genomic analysis to genetically support the metabolic features of the two strains. The genome analysis allowed to identify some gene clusters involved in the metabolism of the main tested xenobiotic compounds. Results show that R7 contains multi- ple genes for the degradation of a large set of aromatic and PAHs compounds, while a lower variability in terms of genes predicted to be involved in aromatic degradation was found in BCP1. This genetic feature can be related to the strong genetic pressure exerted by the two different environment from which the two strains were isolated. According to this, in the BCP1 genome the smo gene cluster involved in the short-chain n-alkanes degradation, is included in one of the unique regions and it is not conserved in the Rhodo- coccus strains compared in this work. Data obtained underline the great potential of these two Rhodococcus spp. strains for biodegradation and environmental decontamination processes

Pool testing on random and natural clusters of individuals: Optimisation of SARS-CoV-2 surveillance in the presence of low viral load samples.

Facing the SARS-CoV-2 epidemic requires intensive testing on the population to early identify and isolate infected subjects. During the first emergency phase of the epidemic, RT-qPCR on nasopharyngeal (NP) swabs, which is the most reliable technique to detect ongoing infections, exhibited limitations due to availability of reagents and budget constraints. This stressed the need to develop screening procedures that require fewer resources and are suitable to be extended to larger portions of the population. RT-qPCR on pooled samples from individual NP swabs seems to be a promising technique to improve surveillance. We performed preliminary experimental analyses aimed to investigate the performance of pool testing on samples with low viral load and we evaluated through Monte Carlo (MC) simulations alternative screening protocols based on sample pooling, tailored to contexts characterized by different infection prevalence. We focused on the role of pool size and the opportunity to develop strategies that take advantage of natural clustering structures in the population, e.g. families, school classes, hospital rooms. Despite the use of a limited number of specimens, our results suggest that, while high viral load samples seem to be detectable even in a pool with 29 negative samples, positive specimens with low viral load may be masked by the negative samples, unless smaller pools are used. The results of MC simulations confirm that pool testing is useful in contexts where the infection prevalence is low. The gain of pool testing in saving resources can be very high, and can be optimized by selecting appropriate group sizes. Exploiting natural groups makes the definition of larger pools convenient and potentially overcomes the issue of low viral load samples by increasing the probability of identifying more than one positive in the same pool