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The mitochondrial Human Proteome Project aims at understanding the function of the mitochondrial proteome and its crosstalk with the proteome of other organelles. Being able to choose a suitable and validated enrichment protocol of functional mitochondria, based on the specific needs of the downstream proteomics analysis, would greatly help the researchers in the field. Mitochondrial fractions from ten model cell lines were prepared using three enrichment protocols and analyzed on seven different LC-MS/MS platforms. All data were processed using neXtProt as reference database. The data are available for the Human Proteome Project purposes through the ProteomeXchange Consortium with the identifier PXD007053. The processed datasets were analysed using a suite of R routines to perform a statistical analysis and to retrieve subcellular and sub-mitochondrial localizations. Although the overall number of identified total and mitochondrial proteins was not significantly dependent on the enrichment protocol, specific line to line differences were observed. Moreover, the protein lists were mapped to a network representing the functional mitochondrial proteome, encompassing mitochondrial proteins and their first interactors. More than 80% of the identified proteins resulted nodes of this network but with a different ability in co-isolating mitochondria-associated structures for each enrichment protocol/cell line pair.

Postpartum haemorrhage (PPH) is one of the main causes of mortality and severe maternal morbidity and its incidence is increasing also in Western countries. Aim of this study is to estimate the incidence and the trend of PPH in the Umbrian population using the validated Umbrian health database and to identify possible determinants for the development of PPH.The source of the data was the regional Healthcare Database of the Umbria Region. The population of interest was represented by women who gave birth in Umbria between 2006 and 2017. The PPH was identified from the hospital data using the ICD-9-CM 666.x codes. Demographic data, principal and secondary diagnoses and data on maternal morbidity and blood component transfusion were collected. The incidence of PPH was calculated taking into account cases of PPH over the total number of births. The determinants of PPH, the associated morbidity and the variation in the severity of the PPH over time have been identified by logistic regression models.In Umbria, between 2006 and 2017, 93,403 births were registered (69% by vaginal delivery and 31% by caesarean section) and the rate of caesarean sections decreased by about 4%. The incidence of PPH increased three-fold during this period with an increase (p0.001) of women with PPH who received transfusions. Regarding the caesarean sections, the PPH trend increased by 53% (p=0.3), while in the vaginal deliveries the PPHs increased by 233% (p0.001). Logistic regression analysis showed that possible risk factors for the occurrence of PPH are maternal morbidity (OR 22.8, 95% CI 18.5-30.0), twin birth (OR 2.0, 95% CI 1.3-3.2) and antepartum haemorrhage (OR 5.7, 95% CI 3.1-10.4).The incidence of PPH has increased in recent years, while the morbidity associated with PPH has remained substantially unchanged. The study identified several risk factors responsible for PPH that can be used in the monitoring of pregnant women and for planning prevention strategies such as Patient Blood Management.

Zollo et al. report that mutations in PRUNE1, a phosphoesterase superfamily molecule, underlie primary microcephaly and profound global developmental delay in four unrelated families from Oman, India, Iran and Italy. The study highlights a potential role for prune during microtubule polymerization, suggesting that prune syndrome may be a tubulinopathy., PRUNE is a member of the DHH (Asp-His-His) phosphoesterase protein superfamily of molecules important for cell motility, and implicated in cancer progression. Here we investigated multiple families from Oman, India, Iran and Italy with individuals affected by a new autosomal recessive neurodevelopmental and degenerative disorder in which the cardinal features include primary microcephaly and profound global developmental delay. Our genetic studies identified biallelic mutations of PRUNE1 as responsible. Our functional assays of disease-associated variant alleles revealed impaired microtubule polymerization, as well as cell migration and proliferation properties, of mutant PRUNE. Additionally, our studies also highlight a potential new role for PRUNE during microtubule polymerization, which is essential for the cytoskeletal rearrangements that occur during cellular division and proliferation. Together these studies define PRUNE as a molecule fundamental for normal human cortical development and define cellular and clinical consequences associated with PRUNE mutation.

Brain-derived neurotrophic factor (BDNF) exerts multiple biological functions in the central nervous system (CNS). It regulates neuronal survival, differentiation and plasticity by activating the receptor TrkB. BDNF levels have been found to be reduced in brain samples of Alzheimer's disease (AD) patients. Alterations in cholesterol homeostasis have also been suggested to play a role in AD. An open question is whether BDNF regulates lipid biosynthesis and metabolism. It was previously reported that BDNF elicits neuronal cholesterol biosynthesis for the development of presynaptic functions in the CNS, but its ability to modulate cholesterol trafficking between astrocytes and neurons was not yet clarified. Major aims of our work were to study BDNF effect on cholesterol efflux and Apolipoprotein E production from astrocytes, and cholesterol incorporation into neurons. The results shows that BDNF significantly increases the synthesis of Apolipoprotein E, which plays a key role in modulating lipid and beta amyloid homeostasis. Also BDNF stimulated cholesterol efflux from astrocytes, by enhancing the expression of ABC transporters. On the other hand, the uptake of cholesterol proteoliposomes in neuronal cells was downregulated by BDNF. These data highlight an additional role of BDNF in cholesterol homeostasis within CNS. A better understanding of the mechanisms by which neurotrophins regulate lipid metabolism is an intriguing issue that will provide a framework for future studies.

Background: Non-pharmacological intervention (e.g. multidisciplinary interventions, music therapy, bright light therapy, educational interventions etc.) are alternative interventions that can be used in older subjects. There are plenty reviews of non-pharmacological interventions for the prevention and treatment of delirium in older patients and clinicians need a synthesized, methodologically sound document for their decision making. Methods and Findings: We performed a systematic overview of systematic reviews (SRs) of comparative studies concerning non-pharmacological intervention to treat or prevent delirium in older patients. The PubMed, Cochrane Database of Systematic Reviews, EMBASE, CINHAL, and PsychINFO (April 28th, 2014) were searched for relevant articles. AMSTAR was used to assess the quality of the SRs. The GRADE approach was used to assess the quality of primary studies. The elements of the multicomponent interventions were identified and compared among different studies to explore the possibility of performing a meta-analysis. Risk ratios were estimated using a random-effects model. Twenty-four SRs with 31 primary studies satisfied the inclusion criteria. Based on the AMSTAR criteria twelve reviews resulted of moderate quality and three resulted of high quality. Overall, multicomponent non-pharmacological interventions significantly reduced the incidence of delirium in surgical wards [2 randomized trials (RCTs): relative risk (RR) 0.71, 95% Confidence Interval (CI) 0.59 to 0.86, I-2=0%; (GRADE evidence: moderate)] and in medical wards [2 CCTs: RR 0.65, 95% CI 0.49 to 0.86, I-2=0%; (GRADE evidence: moderate)]. There is no evidence supporting the efficacy of non-pharmacological interventions to prevent delirium in low risk populations (i.e. low rate of delirium in the control group)[1 RCT: RR 1.75, 95% CI 0.50 to 6.10 ( GRADE evidence: very low)]. For patients who have developed delirium, the available evidence does not support the efficacy of multicomponent non-pharmacological interventions to treat delirium. Among single component interventions only staff education, reorientation protocol (GRADE evidence: very low)] and Geriatric Risk Assessment MedGuide software [hazard ratio 0.42, 95% CI 0.35 to 0.52, (GRADE evidence: moderate)] resulted effective in preventing delirium. Conclusions: In older patients multi-component non-pharmacological interventions as well as some single-components intervention were effective in preventing delirium but not to treat delirium.

We have engineered novel fully human anti-ErbB2 immunoagents: Erbicin, a human scFv (1); ERB-hRNase, a human immunoRNase made up of Erbicin fused to a human RNase (2); ERB-hcAb, a human, “compact” antibody, in which two Erbicin molecules are fused to the Fc of a human IgG1(3,4). Both ERB-hRNase and ERB-hcAb severely inhibit the growth of ErbB2-positive tumours sensitive and resistant to Trastuzumab treatment (5). Furthermore, they are not immunogenic and do not display cardiotoxic effects in vitro and in vivo whereas Trastuzumab was strongly toxic (6). An analysis by three independent methodologies of the interactions of Erbicin-Derived-ImmunoAgents (EDIAs) and Trastuzumab with the ErbB2 extracellular domain (ECD) led to the finding that EDIAs bind soluble ECD with a lower affinity than that for the native receptor on tumor cells. Trastuzumab instead shows a higher affinity for soluble ErbB2-ECD. Accordingly, ErbB2-ECD abolished the biological properties of Trastuzumab with no effects on those of EDIAs (7). These results suggest that the fraction of immunoagent neutralized by free ECD shed into the bloodstream is much higher for Trastuzumab than for EDIAs, thus the latter could be used at lower therapeutic doses compared to those employed with Trastuzumab. More interestingly, we found through the use of three different methods, that the epitope recognized by EDIAs is different from those recognized by Trastuzumab and Pertuzumab, and it is located in the domain I of the extracellular region of ErbB2. This finding could lead to the identification of novel epitopes on ErbB2 to be used as potential therapeutic targets to mitigate anti-ErbB2 associated cardiotoxicity and eventually overcome resistance. 1De Lorenzo, C.; Palmer, DB.; Piccoli, R; Ritter, MA.; and D'Alessio, G.; Clin. Cancer Res. 2002, 8, 1710-1719. 2De Lorenzo, C.; Arciello, A.; Cozzolino, R.; Palmer, DB.; Laccetti, P.; Piccoli, R; and D'Alessio, G.; Cancer Res. 2004, 64, 4870-4874. 3De Lorenzo, C.; Tedesco, A.; Terrazzano, G.; Cozzolino, R.; Laccetti, P.; Piccoli, R.; and D'Alessio, G.; Br. J. Cancer 2004, 91, 1200-1204. 4De Lorenzo, C.; Cozzolino, R.; Carpentieri, A.; Pucci, P.; Laccetti, P.; and D'Alessio, G.; Carcinogenesis 2005, 26 ,1890-1895. 5 Gelardi, T.; Damiano, V.; Rosa, R..; Bianco, R..; Cozzolino, R.; Tortora, G.; Laccetti, P.; D'Alessio, G.; and De Lorenzo, C; Br. J. Cancer 2010, 102, 513-519. 6 Riccio, G.; Esposito, G.; Leoncini, E.; Contu, R..; Condorelli, G.; Chiariello, M.; Laccetti, P.; Hrelia, S.; D'Alessio, G.; and De Lorenzo, C.; Faseb J. 2009, 23, 3171-3178 7Troise, F.; Cafaro,V.; Giancola, C.; D’Alessio G.; and De Lorenzo, C.; FEBS J. 2008, 275, 4967–4979.

Two novel human antitumor immunoconjugates, engineered by fusion of a single-chain antibody fragment against human ErbB2 receptor, termed Erbicin, with either a human RNase or the Fc region of a human IgG(1) , are selectively cytotoxic for ErbB2-positive cancer cells in vitro and in vivo. These Erbicin-derived immunoagents (EDIAs) do not show the most negative properties of Herceptin, the only humanized mAb against ErbB2 used in the therapy of breast carcinoma: cardiotoxicity and the inability to act on resistant tumors. These differences are probably attributable to the different ErbB2 epitopes recognized by EDIAs and Herceptin, respectively, as we have previously reported that they induce different signaling mechanisms that control tumor and cardiac cell viability. Thus, to accurately identify the novel epitope recognized by EDIAs, three independent and complementary methodologies were used. They gave coherent results, which are reported here: EDIAs bind to a different ErbB2 epitope than Herceptin and the other human/humanized antibodies against ErbB2 reported so far. The epitope has been successfully located in region 122-195 of extracellular domain I. These findings could lead to the identification of novel epitopes on ErbB2 that could be used as potential therapeutic targets to mitigate anti-ErbB2-associated cardiotoxicity and eventually overcome resistance.

Oral Lichen Planus is a chronic inflammatory pathology with not well defined etiology, characterized by an immunoreactivity directed against the keratinocytes of basal layer and mediated by a cellular infiltrate of T-lymphocytes. This immunoreactivity is caused by a modification of basal keratinocytes surface antigens, with activation of antigen-presenting cells allowing the recognition on non-self antigens by CD4 lymphocytes. The migration of T-cells at the sites of the damage and their interactions with leucocytes and keratinocytes represent a key effect of pathogenesis of OLP, and are mediated by specific cell surface adhesion molecules. Recent studies in molecular biology, with the discovery and the definition of cell adhesion receptors, have greatly contributed to clarify the pathogenic mechanism of OLP; these molecules regulate immune functions such as adhesion to endothelial cells, lymphocytes migration into extravasal tissues at the site of immune response and T-cell interactions with target cell antigens. The adhesion molecules have been classified into various families, differing on the basis of their molecular structure: the Immunoglobulin Superfamily (ICAM 1-3, PECAM-1, VCAM-1), the Selectins (ELAM-1, LECAM-1, GMP-140), the Integrins (LEA-1) and others. In this paper, the principal classes of adhesion molecules which are involved in the immune response are described and their importance in the pathogenesis of Lichen Planus is underlined.