104 research outputs found
Base excision repair intermediates are mutagenic in mammalian cells
Base excision repair (BER) is the main pathway for repair of DNA damage in mammalian cells. This pathway leads to the formation of DNA repair intermediates which, if still unsolved, cause cell lethality and mutagenesis. To characterize mutations induced by BER intermediates in mammalian cells, an SV-40 derived shuttle vector was constructed carrying a site-specific lesion within the recognition sequence of a restriction endonuclease. The mutation spectra of abasic (AP) sites, 5âČ-deoxyribose-5-phosphate (5âČdRp) and 3âČ-[2,3-didehydro-2,3-dideoxy-ribose] (3âČddR5p) single-strand breaks (ssb) in mammalian cells was analysed by RFLP/PCR and mutation frequency was estimated by quantitative PCR. Point mutations were the predominant events occurring at all BER intermediates. The AP site-induced mutation spectrum supports evidence for the âA-ruleâ and is also consistent with the use of the 5âČ neighbouring base to instruct nucleotide incorporation (5âČ-rule). Preferential adenine insertion was also observed after in vivo replication of 5âČdRp or 3âČddR5p ssb. We provide original evidence that not only the abasic site but also its derivatives âfacelessâ BER intermediates are mutagenic, with a similar mutation frequency, in mammalian cells. Our findings support the hypothesis that unattended BER intermediates could be a constant threat for genome integrity as well as a spontaneous source of mutations
Drug addicted mothers and their empathic reactivity to painful cues.
Introduction: Mother-infant relationship is crucial for offspring\u2019s development. Substance-abuse has been shown to affect adults\u2019 ability to care for their children, in particular in the affective- relational aspects of parenting, such as the attachment bond and the ability to provide protection. Surprisingly, no previous study has explored neural responses associated with empathy towards children\u2019s pain in mothers with such history. Empathy is a more general construct than parenting and deficits in neural empathic responses may better explain failures in caring of addicted
mothers.
Aims of the study: This study was aimed at investigating the empathic response to pain inflicted by a dangerous tool to hands of kids when compared to neutral situations in a sample of mothers with history of drug-abuse. We used Event-Related Potentials (ERPs) technique to explore the time-course of neural empathic responses to pain by virtue of its excellent time resolution. In particular, we asked whether empathic responses to pain of children would differ in drug-addicted mothers compared to control group, during a relatively automatic early stage of processing or during a more controlled delayed one involving mentalizing.
Material and methods: Event-related potentials (ERPs) have been recorded from drug-addicted mothers (i.e., clinical) and control groups to track neural activity in a pain decision task. Stimuli were pictures showing one infant hand in painful (harmful object applied to the hand) and neutral situations (harmful object located close to the hand).
Results: Neural empathic responses towards children in pain differed between the two groups such that ERPs diverged between the painful and neutral stimulation at delayed stages of processing only for the clinical group.
Conclusions: We interpreted these results as indicating that the control group implicitly judged also the neutral situations involving children as potentially painful supporting a lack of mentalizing abilities in the clinical group when compared with controls
Evaluation by environmental monitoring of pesticide absorption in farm workers of 18 Italian tomato cultivations
Tornato cultivation farms of Soutbern Italy were investigated in order to evaluate tbe generai working conditions and tbe levels of exposure of farm workers to pesticides, during tbe mixinglloading and tbe application of pesticides on fields. Information on working modalities, personal protective equipment, etc. was collected using a questionnaire. Inbaling and cutaneous exposure levels were measured, and tbe estimated pesticide total absorbed dose was compared witb Admissible Daily Intakes (ADIs). Field treatments were mainly carried out by using sprayers witb open cab tractors, and, in 57 ~9% of cases, tbe pesticide mixture was manually prepared by mixing pesticides in a paU, often witbout using gloves (59.5%). Tbe estimated pesticides absorbed doses varied in tbe range 0.56-2630.31 mg (mean value, 46.9 mg), and 20% of tbe measured absorbed doses exceeded ADIs. Tbe findings obtained in tbe 18 examined farms sbow a worrying situation, suggesting tbe investigation of many more farms, so tbat a statistically significant picture of tornato cultivations in Soutbern Italy could be formed. Besides, tbe planning of training courses aimed to increase workers consciousness about bealtb risks and bow tbey can be prevented is advisable
Triple Vectors Expand AAV Transfer Capacity in the Retina.
Retinal gene transfer with adeno-associated viral (AAV) vectors holds great promise for the treatment of inherited retinal degenerations (IRDs). One limit of AAV is its transfer capacity of about 5 kb, which can be expanded to about 9 kb, using dual AAV vectors. This strategy would still not suffice for treatment of IRDs such as Usher syndrome type 1D or Alström syndrome type I (ALMS) due to mutations in CDH23 or ALMS1, respectively. To overcome this limitation, we generated triple AAV vectors, with a maximal transfer capacity of about 14 kb. Transcriptomic analysis following triple AAV transduction showed the expected full-length products along a number of aberrant transcripts. However, only the full-length transcripts are efficiently translated in vivo. We additionally showed that approximately 4% of mouse photoreceptors are transduced by triple AAV vectors and showed correct localization of recombinant ALMS1. The low-photoreceptor transduction levels might justify the modest and transient improvement we observe in the retina of a mouse model of ALMS. However, the levels of transduction mediated by triple AAV vectors in pig retina reached 40% of those observed with single vectors, and this bodes well for further improving the efficiency of triple AAV vectors in the retina. Mol Ther 2018 Feb 7 26(2):524-541
Controlled degradability of PCL-ZnO nanofibrous scaffolds for bone tissue engineering and their antibacterial activity
Up to date, tissue regeneration of large bone defects is a clinical challenge under exhaustive study. Nowadays, the most common clinical solutions concerning bone regeneration involve systems based on human or bovine tissues, which suffer from drawbacks like antigenicity, complex processing, low osteoinductivity, rapid resorption and minimal acceleration of tissue regeneration. This work thus addresses the development of nanofibrous synthetic scaffolds of polycaprolactone (PCL) - a long-term degradation polyester - compounded with hydroxyapatite (HA) and variable concentrations of ZnO as alternative solutions for accelerated bone tissue regeneration in applications requiring mid- and long-term resorption. In vitro cell response of human fetal osteoblasts as well as antibacterial activity against Staphylococcus aureus of PCL:HA:ZnO and PCL:ZnO scaffolds were here evaluated. Furthermore, the effect of ZnO nanostructures at different concentrations on in vitro degradation of PCL electrospun scaffolds was analyzed. The results proved that higher concentrations ZnO may induce early mineralization, as indicated by high alkaline phosphatase activity levels, cell proliferation assays and positive Alizarin-Red-S-stained calcium deposits. Moreover, all PCL:ZnO scaffolds particularly showed antibacterial activity against S. aureus which may be attributed to release of Zn2+ ions. Additionally, results here obtained showed a variable PCL degradation rate as a function of ZnO concentration. Therefore, this work suggests that our PCL:ZnO scaffolds may be promising and competitive short-, mid- and long-term resorption systems against current clinical solutions for bone tissue regeneration.Fil: Felice, Betiana. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas. Universidad Nacional de TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas; Argentina. Universidad Nacional de TucumĂĄn. Facultad de Ciencias Exactas y TecnologĂa; ArgentinaFil: Sanchez, Maria Alejandra. Universidad Nacional de TucumĂĄn. Facultad de Ciencias Exactas y TecnologĂa; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas. Universidad Nacional de TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas; ArgentinaFil: Socci, Maria Cecilia. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas. Universidad Nacional de TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas; Argentina. Universidad Nacional de TucumĂĄn. Facultad de Ciencias Exactas y TecnologĂa; ArgentinaFil: Sappia, Luciano David. Universidad Nacional de TucumĂĄn. Facultad de Ciencias Exactas y TecnologĂa; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas. Universidad Nacional de TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas; ArgentinaFil: GĂłmez, MarĂa InĂ©s. Universidad Nacional de TucumĂĄn. Facultad de BioquĂmica, QuĂmica y Farmacia. Instituto de QuĂmica InorgĂĄnica; ArgentinaFil: Cruz, MarĂa Karina. Universidad Nacional de TucumĂĄn. Facultad de BioquĂmica, QuĂmica y Farmacia. Instituto de QuĂmica InorgĂĄnica; ArgentinaFil: Felice, Carmelo Jose. Universidad Nacional de TucumĂĄn. Facultad de Ciencias Exactas y TecnologĂa; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas. Universidad Nacional de TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas; ArgentinaFil: MartĂ, MercĂš. Universitat AutĂČnoma de Barcelona; EspañaFil: Pividori, MarĂa Isabel. Universitat AutĂČnoma de Barcelona; EspañaFil: Simonelli, Gabriela. Universidad Nacional de TucumĂĄn. Instituto de FĂsica del Noroeste Argentino. - Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet Noa Sur. Instituto de FĂsica del Noroeste Argentino; ArgentinaFil: Rodriguez, Andrea Paola. Universidad Nacional de TucumĂĄn. Facultad de Ciencias Exactas y TecnologĂa; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas. Universidad Nacional de TucumĂĄn. Instituto Superior de Investigaciones BiolĂłgicas; Argentin
IgE antibody repertoire in nasal secretions of children and adults with seasonal allergic rhinitis: A molecular analysis
Background:
There is growing interest both in testing IgE in nasal secretions (NS) and in molecular diagnosis of seasonal allergic rhinitis (SAR). Yet, the reliability of nasal IgE detection with the newest molecular assays has never been assessed in a large cohort of pollen allergic patients.
Objective:
To investigate with microarray technology and compare the repertoires of specific IgE (sIgE) antibodies in NS and sera of a large population of children and adults with SAR.
Methods:
Nasal secretions were collected with an absorbent device (Merocel 2000Âź, Medtronic) and a minimal dilution procedure from 90 children and 71 adults with SAR. Total IgE (tIgE) (ImmunoCAP, Thermo Fisher Scientific (TFS)) and sIgE antibodies against 112 allergen molecules (ISAC-112, TFS) were measured in NS and serum.
Results:
Nasal sIgE was detectable in 68.3% of the patients. The detected nasal sIgE antibodies recognized airborne (88%), vegetable (10%), and animal food or other (<1%) allergen molecules. The prevalence and average levels of sIgE in NS and serum were highly interrelated at population level. A positive nasal sIgE antibody to a given molecule predicted the detection of the same antibody in the patient's serum with a specificity of 99.7% and a sensitivity of 40%.
Conclusions:
The concentration of sIgE is much lower in nasal secretions than in the serum. sIgE assays with very high analytical sensitivity and sampling methods with minimal dilution will be therefore needed to validate nasal secretions as alternative to serum in testing the sIgE repertoire
Intraperitoneal drain placement and outcomes after elective colorectal surgery: international matched, prospective, cohort study
Despite current guidelines, intraperitoneal drain placement after elective colorectal surgery remains widespread. Drains were not associated with earlier detection of intraperitoneal collections, but were associated with prolonged hospital stay and increased risk of surgical-site infections.Background Many surgeons routinely place intraperitoneal drains after elective colorectal surgery. However, enhanced recovery after surgery guidelines recommend against their routine use owing to a lack of clear clinical benefit. This study aimed to describe international variation in intraperitoneal drain placement and the safety of this practice. Methods COMPASS (COMPlicAted intra-abdominal collectionS after colorectal Surgery) was a prospective, international, cohort study which enrolled consecutive adults undergoing elective colorectal surgery (February to March 2020). The primary outcome was the rate of intraperitoneal drain placement. Secondary outcomes included: rate and time to diagnosis of postoperative intraperitoneal collections; rate of surgical site infections (SSIs); time to discharge; and 30-day major postoperative complications (Clavien-Dindo grade at least III). After propensity score matching, multivariable logistic regression and Cox proportional hazards regression were used to estimate the independent association of the secondary outcomes with drain placement. Results Overall, 1805 patients from 22 countries were included (798 women, 44.2 per cent; median age 67.0 years). The drain insertion rate was 51.9 per cent (937 patients). After matching, drains were not associated with reduced rates (odds ratio (OR) 1.33, 95 per cent c.i. 0.79 to 2.23; P = 0.287) or earlier detection (hazard ratio (HR) 0.87, 0.33 to 2.31; P = 0.780) of collections. Although not associated with worse major postoperative complications (OR 1.09, 0.68 to 1.75; P = 0.709), drains were associated with delayed hospital discharge (HR 0.58, 0.52 to 0.66; P < 0.001) and an increased risk of SSIs (OR 2.47, 1.50 to 4.05; P < 0.001). Conclusion Intraperitoneal drain placement after elective colorectal surgery is not associated with earlier detection of postoperative collections, but prolongs hospital stay and increases SSI risk
I disturbi della relazione come nuove entitaÌ cliniche nella diagnosi e nell'intervento
In questo capitolo, in continuitaÌ con unâidea di psicopatologia evolutiva che pone lâattenzione sulla dimensione relazionale, si affronta la questione della diagnosi psicopatologica in etaÌ evolutiva, attraverso il riferimento specifico agli strumenti di classificazione della psicopatologia stessa: il Manuale Diagnostico Statistico dei disturbi mentali (DSM) nella sua quarta edizione (DSM-IV-Tr; APA, 2000), lâInternational Classification of Diseases (ICD-10, WMH, 1992) alla sua decima edizione e la Classificazione Diagnostica 0-3 (Zero-to-Three, 1994) di recente introduzione; sistemi che si basano su un approccio che cerca di essere ateoretico, cioeÌ svincolato da un modello concettuale di riferimento rispetto alle cause (eziologia) e al processo fisiopatologico allâorigine del disturbo, teso ad identificare categorie, ovvero sindromi omogenee e distinte, e fondato su criteri diagnostici che permettono di rilevare i sintomi ricorrendo ad un livello di inferenza minimo. La discussione condotta eÌ orientata ad evidenziare che i sistemi/strumenti diagnostici appena indicati, pur proponendosi un approccio innovativo alla classificazione psicopatologica in etaÌ evolutiva, non riescono a riconoscere adeguatamente alcuni aspetti importanti dal punto di vista dello sviluppo, come, per esempio la centralitaÌ della relazione caregiver-bambino nella strutturazione e nel mantenimento della patologia. Ne deriva la necessitaÌ di porre lâenfasi sulla possibilitaÌ di comprendere lâinterazione dinamica tra le caratteristiche del bambino e quelle dellâadulto nella relazione, cosiÌ come sulle influenze della qualitaÌ della relazione stessa sui funzionamenti individuali. Secondo tale prospettiva, lo sforzo teorico e metodologico nella descrizione e nella classificazione dei disturbi in etaÌ evolutiva deve riguardare la possibilitaÌ di determinare se i disturbi della relazione caregiver-bambino siano semplicemente caratteristiche associate o, piuttosto, parte integrante del disturbo dello sviluppo del bambino. In questo senso, sebbene i sistemi di classificazione diagnostica piuÌ recenti abbiano compiuto notevoli sforzi per integrare piuÌ di un focus relazionale, riuscire ad articolare e ridefinire un sistema di classificazione appropriato al modello della relazione resta una sfida tuttora in corso. Sulla base di questo approccio vengono pertanto dimostrati i limiti nella classificazione della psicopatologia evolutiva secondo il DSM-IV-Tr e anche secondo la classificazione O-3. Lâultima parte del capitolo pone lâenfasi su un orientamento transazionale che riconosce la mutua influenza di entrambi i soggetti coinvolti nella relazione, collocando il disturbo nella relazione specifica tra i due soggetti stessi, e sviluppando in modo particolare le diverse tipologie di intervento (riparazione, ridefinizione, rieducazione), in unâottica teorica centrata su unâidea di relazione come costrutto globale
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