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Supplementary Figure 1 from The Tyrosine Phosphatase Shp2 Interacts with NPM-ALK and Regulates Anaplastic Lymphoma Cell Growth and Migration

Anaplastic large cell lymphomas (ALCL) are mainly characterized by the reciprocal translocation t(2;5)(p23;q35) that involves the anaplastic lymphoma kinase (ALK) gene and generates the fusion protein NPM-ALK with intrinsic tyrosine kinase activity. NPM-ALK triggers several signaling cascades, leading to increased cell growth, resistance to apoptosis, and changes in morphology and migration of transformed cells. To search for new NPM-ALK interacting molecules, we developed a mass spectrometry–based proteomic approach in HEK293 cells expressing an inducible NPM-ALK and identified the tyrosine phosphatase Shp2 as a candidate substrate. We found that NPM-ALK was able to bind Shp2 in coprecipitation experiments and to induce its phosphorylation in the tyrosine residues Y542 and Y580 both in HEK293 cells and ALCL cell lines. In primary lymphomas, antibodies against the phosphorylated tyrosine Y542 of Shp2 mainly stained ALK-positive cells. In ALCL cell lines, Shp2-constitutive phosphorylation was dependent on NPM-ALK, as it significantly decreased after short hairpin RNA (shRNA)–mediated NPM-ALK knock down. In addition, only the constitutively active NPM-ALK, but not the kinase dead NPM-ALKK210R, formed a complex with Shp2, Gab2, and growth factor receptor binding protein 2 (Grb2), where Grb2 bound to the phosphorylated Shp2 through its SH2 domain. Shp2 knock down by specific shRNA decreased the phosphorylation of extracellular signal-regulated kinase 1/2 and of the tyrosine residue Y416 in the activation loop of Src, resulting in impaired ALCL cell proliferation and growth disadvantage. Finally, migration of ALCL cells was reduced by Shp2 shRNA. These findings show a direct involvement of Shp2 in NPM-ALK lymphomagenesis, highlighting its critical role in lymphoma cell proliferation and migration. [Cancer Res 2007;67(9):4278–86]

The seagrass Halophila stipulacea is native to the Red Sea. It invaded the Mediterranean over the past century and most of the Caribbean over the last two decades. Understanding the main drivers behind the successful invasiveness of H. stipulacea has become crucial. We performed a comprehensive study including field measurements, a mesocosm experiment, and a literature review to identify ‘superior traits’ that can potentially explain the success story of H. stipulacea. We assessed meadow characteristics and plant traits of three invasive H. stipulacea populations growing off the Island of Sint Eustatius (eastern Caribbean). We compared similar parameters between native (Eilat, Red Sea) and invasive H. stipulacea plants in a common-garden mesocosm. Lastly, we also compared our field measurements with published data. The newly arrived H. stipulacea plants from St. Eustatius were characterized by higher percent cover, higher below- and above-ground biomasses, more apical shoots, and faster leaf turnover rates than those measured in both native and older invaded habitats. These results were further confirmed by mesocosm experiments where the invasive H. stipulacea plants grew faster and developed more apical shoots than the native plants. Results suggest that increased growth vigour is one of the main invasive traits that characterize successful invasive H. stipulacea populations in the Caribbean and potentially in other invaded areas. We encourage long-term monitoring of H. stipulacea in both native and invaded habitats to better understand the future spread of this species and its impacts on communities and their ecosystem functions and services.

Background Breast cancer (BC) is the most commonly diagnosed cancer and the second leading cause of cancer-related deaths among women in Africa after cervical cancer. Even if the epidemiological data are now aligned with those relating to industrialized countries, the knowledge concerning breast cancer in Africa, particularly in Western Africa, still lack clinical data, medical treatments, and the evaluation of genetic and non-genetic factors implicated in the etiology of the disease. The early onset and the aggressiveness of diagnosed breast cancers in patients of African ancestry strongly suggest that the genetic risk factor may be a key component, but so far, very few studies on the impact of germ line mutations in breast cancer in Africa have been conducted, with negative consequences on prevention, awareness and patient management. Through Next Generation sequencing (NGS), we analyzed all of the coding regions and the exon–intron junctions of BRCA1 and BRCA2 genes—the two most important genes in hereditary breast cancer—in fifty-one women from Burkina Faso with early onset of breast cancer with or without a family history. Results We identified six different pathogenic mutations (three in BRCA1, three in BRCA2), two of which were recurrent in eight unrelated women. Furthermore, we identified, in four other patients, two variants of uncertain clinical significance (VUS) and two variants never previously described in literature, although one of them is present in the dbSNP database. Conclusions This is the first study in which the entire coding sequence of BRCA genes has been analyzed through Next Generation Sequencing in Burkinabe young women with breast cancer. Our data support the importance of genetic risk factors in the etiology of breast cancer in this population and suggest the necessity to improve the genetic cancer risk assessment. Furthermore, the identification of the most frequent mutations of BRCA1 and BRCA2 in the population of Burkina Faso will allow the development of an inexpensive genetic test for the identification of subjects at high genetic cancer risk, which could be used to design personalized therapeutic protocols.

Background: Breast cancer (BC) is the most commonly diagnosed cancer and the second leading cause of cancer-related deaths, after cervical cancer, among women in Africa. Even if the epidemiological data are now aligned with those relating to industrialized countries, the knowledge concerning breast cancer in Africa, in particularly in Western Africa still lack clinical data, medical treatments, and the evaluation of genetic and non-genetic factors implicated in the etiology of the disease.The early onset and the aggressiveness of diagnosed breast cancers in patients of African ancestry strongly suggest that the genetic risk factor may play an important role but up to date very few studies have been done concerning the impact of germ line mutations in breast cancer in Africa, with a negative impact on prevention, awareness and patient management.We have performed by Next Generation sequencing (NGS) the analysis of all coding regions and the exon-intron junctions of BRCA1 and BRCA2 genes, the two most important genes in hereditary breast cancer, in fifty-one women from Burkina Faso with early onset of breast cancer and or without a family history.Results: We identified six different pathogenic mutations (3 in BRCA1, 3 in BRCA2), two of which have been found to be recurrent , in 8 unrelated women.In addition, we identified, in 4 other patients, two variants of uncertain clinical significance (VUS) and two variants never previously described in literature, although one of which is present in the dbSNP database.Conclusions: The present study is the first in which the entire coding sequence of BRCA genes have been analyzed by Next Generation Sequencing in Burkinabe young women with breast cancer. Our data support the importance of genetic risk factors in the etiology of breast cancer in this population and suggests the necessity to improve the genetic cancer risk assessment. Furthermore, the identification of the most frequent mutations of BRCA1 and BRCA2 in the population of Burkina Faso will allow the development of an inexpensive genetic test for the identification of subjects at high genetic cancer risk, which could be used to design personalized therapeutic protocols.

Seagrasses and associated microbial communities constitute a functional unit (holobiont) which responds as a whole to environmental changes. However, it is still unclear how the microbial colonizers are selected. In this study we compared the epiphytic microbial communities associated with Posidonia oceanica and Halophila stipulacea, Mediterranean native and exotic seagrass species, respectively, growing side by side in monospecific patches within the port of Limassol (Cyprus, Eastern Mediterranean Sea). To evaluate whether the environment rather than the host species and/or its physiological condition play a role in shaping the seagrass epiphytic microbial community, the environmental microbial communities (seawater and sediment) and seagrass associated ones were determined by using 16S rRNA gene amplicon sequencing. Plant ecological status was evaluated by morphological (biometry), structural (density) and biochemical (pigment/phenol content) descriptors. In both species, leaf associated microbial communities are clearly similar to seawater microbes; conversely, microbes associated with H. stipulacea roots/rhizomes differ from the microbial communities in surrounding sediment. In both seagrasses, Pseudomonadaceae was the most abundant family on leaves, but each species harboured unique microbial families. To our best knowledge, this is the first study on these two neighbouring seagrass species, coupling plant ecological status with associated microbial communities. Results demonstrated that each seagrass responded differently to the same environmental conditions and selected different epiphytic microbial communities, supporting their putative use as ecological indicators.

The microbial communities associated with plant's compartments and the expression of the chloroplast chaperonin Hsp60 have been simultaneously analyzed during the diel cycle in the marine seagrass Thalassia hemprichii from the lagoon of Magoodhoo island (Maldives), characterized by remarkable daily shifts in temperature and light. Plants showed a significant up-regulation of Hsp60 expression from 8.00 a.m. to 2.00 p.m., in correspondence with the increase in temperature and light, confirming their role as defense mechanism against photoinhibition and oxidative damage. However, a further significant increase of the Hsp60 level was also observed when irradiance and temperature dropped, suggesting that the cellular stress was still in progress. The plant-associated microbial communities showed differences by plant compartment and sampling time, with the aboveground compartment (leaves) being much more dynamic than the belowground one (roots/rhizomes). In the phyllosphere, a progressive shift during the day from the absolute dominance of the Gammaproteobacteria class, mainly constituted of Enterobacteraceae family, to the increase of the biodiversity due to the rise of Alphaproteobacteria was observed. Belowground, the microbial diversity was much lower than aboveground, being Gammaproteobacteria the most represented class throughout all sampling times. Vibrionaceae family was the most abundant at 8:00 a.m. and 6.00 p.m. decreasing slightly at 2.00 p.m., partially replaced by Halomonadaceae. The combined biochemical and microbial markers allows to assess the plant stress response and to deepen the knowledge on the seagrasses adaptation to harsh and changing environmental conditions, resulting useful to detect early signs of change in an organism's physiological state. Furthermore, the variation of Hsp60 expression and associated bacterial communities in response to light and temperature fluctuations support the ‘holobiont’ theory, which considers the plant/microorganisms association as a functional unit, as suggested for corals.

One major issue in the aftermath of a strong earthquake is the emergency management, which cannot be guaranteed if the physical components necessary for the operability of the contingency plan are either damaged, unusable or inaccessible. In order to assess the physical efficiency of the emergency system on which a contingency plan is based, the I.OPà.CLE method (Indices for evaluation of the Operational efficiency of Limit Condition Emergency) has been set up by the Italian Civil Protection Department. It is basically aimed at supporting civil protection decision makers in evaluating emergency systems and establishing strategies and priorities concerning strengthening interventions at national, regional or municipality level. The method must be potentially applied to any of the around 8000 municipalities all over Italy, and then it must refer to a minimum information dataset readily available for any municipality. Therefore, I.OPà.CLE has been specifically tailored for the minimum standard information that is provided by the analysis of Limit Condition for Emergency, as formulated in 2012 by the Italian Civil Protection Department. This analysis defines the standard framework of the physical layout of a municipal seismic emergency plan, relying on a limited number of information. Notwithstanding the numerically extensive target and limited amount of information available, the ambition of I.OPà.CLE is to perform simplified analyses without renouncing to rigor in the methodology. The present paper illustrates the analytic formulation of I.OPà.CLE and its capabilities, as well as two real examples. In the conclusion, future upgrading of the model are discussed, even in relation to the availability of further pieces of information.

The Italian National Seismic Prevention Plan (NSPP) was issued following the destructive 2009 L'Aquila earthquake, with a total endowment of 965 million euros granted by law, under the coordination of the Italian Civil Protection Department (ICPD). The NSPP addresses seismic risk reduction according to a broad view, through multi-task programs. Among the measures granted by NSPP, one is devoted to the seismic retrofit of strategic or critical public buildings and infrastructures, whose damage or collapse may affect emergency management as well as produce high consequences in terms of human lives. The paper provides a preliminary assessment of such seismic prevention measure, through some statistical elaborations on a sample of around 1000 interventions, 375 of which so far concluded. The analyses carried out on this sample provide interesting information to verify the effectiveness of the measure as well as to enhance further actions similarly purposed. Analyses on seismic risk indices of critical buildings included in the NSPP are presented and discussed in the paper, together with an early analysis of costs and benefits associated with different structural types, and intervention types. The results obtained highlight interesting aspects in terms of seismic safety increases and marginal costs for the masonry and reinforced concrete buildings of the considered sample. These preliminary results, though requiring further calibrations on the larger datasets foreseen in the future, provide useful hints to envisage how they could be used in the definition of future strategies for seismic risk reduction. To this aim, a final paragraph provides an exemplification and a simplified procedure based on the findings shown in the paper, for comparing costs and benefits of different strategies aimed at reducing the risk of a given portfolio of buildings.

The procedure for pump qualification and verification for the European market is now in force. In this paper, tests performed in the Hydro-Energy Laboratory (HELab), at the University of Naples Federico II in Italy, are described and the results discussed.

Background Marfan Syndrome (MFS) is a chronic, life-threatening, autosomal dominant connective tissue disorder caused by mutations in the FBN1 gene, coding for fibrillin-1. All organ systems may be affected, but particularly the cardiovascular system, eyes, and skeleton. Mortality generally results from cardiovascular complications, mainly aortic dissection. Currently, the diagnosis of MFS is based on the revised Ghent nosology. Molecular analysis of the FBN1 gene reduces diagnostic uncertainty in patients with suspected MFS or MFS-related disorders (MFS-RD). To date, more than 2700 FBN1 mutations are known. Methods Using Next Generation Sequencing (NGS) followed by Multiplex Ligation-dependent Probe Amplification on NGS-negative samples, we screened FBN1 gene on 124 unrelated patients (101 MFS fulfilling revised Ghent criteria, 20 suspected MFS, 3 MFS-RD) enrolled from 2008 to 2018 at the Multidisciplinary Marfan Clinic, Tor Vergata Hospital, Rome. Results An FBN1 variant was identified in 107/124 (86.3%) patients, including 48 novel variants (46 pathogenic/likely pathogenic, 2 VUS). A pathogenic/likely pathogenic variant was detected in 90/101 (89.1%) MFS patients. Our approach allowed early diagnosis for 10 young patients (age 3–19 years) with suspected MFS. Conclusions This study broadens the mutation spectrum of FBN1, providing a full update of the molecular basis of MFS in Italy.

A seguito del terremoto che ha colpito la Regione Abruzzo il 6 aprile 2009, il Governo Italiano ha emanato un nuovo provvedimento per dare maggiore impulso alla prevenzione sismica. L'articolo 11 del decreto legge n. 39 del 28 aprile 2009 prevede che siano finanziati interventi per la prevenzione del rischio sismico su tutto il territorio nazionale sancendo in questo modo l’avvio di un Piano Nazionale di prevenzione sismica. Il Fondo istituito allo scopo, 965 milioni di euro ripartiti in 7 anni, rappresenta tuttavia solo una minima percentuale del fabbisogno necessario in Italia per la mitigazione del rischio. La gestione del Piano è stata affidata al Dipartimento della Protezione Civile, attraverso il coinvolgimento a livello territoriale delle amministrazioni regionali, responsabili dell’attuazione sul territorio dei programmi di prevenzione. Tra le linee di finanziamento avviate, una di esse è dedicata agli interventi di riqualificazione sismica di edifici di interesse strategico e delle opere infrastrutturali la cui funzionalità durante gli eventi sismici assume rilievo fondamentale per le finalità di protezione civile, nonché degli edifici e delle opere rilevanti in relazione alle conseguenze di un collasso. Ad oggi il numero complessivo di interventi programmati tra quelli finanziati (rafforzamento locale, miglioramento sismico, demolizione e ricostruzione) ammonta a circa 1000, ma si prevede che termine del Piano questo numero cresca ulteriormente. A dieci anni dal provvedimento, si ritiene utile produrre un quadro di sintesi di quanto ad oggi realizzato nell’ambito di tale misura, attraverso alcune elaborazioni statistiche sul campione di interventi disponibile. L’articolo, in particolare, indaga la vulnerabilità e il deficit di sicurezza degli edifici strategici e rilevanti del campione, insieme a una prima analisi dei costi e dei tempi medi necessari per la realizzazione dei relativi interventi, con l'obiettivo finale di fornire informazioni utili anche per meglio indirizzare le future attività di prevenzione strutturale.

New experimental data on pressure-pipe flow of slurries, together with some literature data, are used to evaluate the performance of four resistance models based on different interpretations of the process. The experiments were performed on a mixture of water and uniform sediment showing a shear-thinning rheological behavior. The literature datasets refer to a Bingham plastic mixture and to a shear-thickening mixture. Two of the tested models assume a fully stratified flow and express the total resistances as the sum of the bed and of the vehicle contribution. The third and the fourth models are based on a rheological approach that considers an equivalent homogenous fluid.

Mutations within EDA1 gene, which encodes for the ectodysplasin, cause X-linked anhidrotic ectodermal dysplasia. In this study, 23 Italian patients with anhidrotic ectodermal dysplasia were analyzed for mutations in EDA1 gene. We set up a rapid protocol through denaturing high-performance liquid chromatography, followed by sequencing, that allowed the characterization of 18 mutations, 14 novel and 4 recurrent: 8 missense mutations (p.L51Q, p.H54R, p.R156H twice, p.C332F, p.D316H, p.T378M, and p.A349T), 3 in-frame deletions (p.G82_P84del, p.A179_P191del, and p.L354del), 1 gross deletion (p.G168_G265del, identified through direct sequencing and PCR), 4 altered splicing (c.949-13T > C, c.741 + 1G/T, c.793 + 4A > T, and c.924 + 1G/T), 1 nonsense (p.Y3X), and 1 synonymous mutation (c.741G > A). Moreover, structural analysis of three missense mutations shows that alteration of the electrostatic surface of the protein (p.D316N), the break of intermonomer interactions (p.A349T) and destabilization of the single monomer structure (p.T378M), may irreversibly invalidate the EDA-A1 binding properties. Our data confirm and extend the large spectrum of EDA1 mutations and provide a rapid and efficient molecular protocol for testing EDA1 mutations in EDA patients.

Anaplastic large cell lymphomas (ALCL) are mainly characterized by the reciprocal translocation t(2;5)(p23;q35) that involves the anaplastic lymphoma kinase (ALK) gene and generates the fusion protein NPM-ALK with intrinsic tyrosine kinase activity. NPM-ALK triggers several signaling cascades, leading to increased cell growth, resistance to apoptosis, and changes in morphology and migration of transformed cells. To search for new NPM-ALK interacting molecules, we developed a mass spectrometry–based proteomic approach in HEK293 cells expressing an inducible NPM-ALK and identified the tyrosine phosphatase Shp2 as a candidate substrate. We found that NPM-ALK was able to bind Shp2 in coprecipitation experiments and to induce its phosphorylation in the tyrosine residues Y542 and Y580 both in HEK293 cells and ALCL cell lines. In primary lymphomas, antibodies against the phosphorylated tyrosine Y542 of Shp2 mainly stained ALK-positive cells. In ALCL cell lines, Shp2-constitutive phosphorylation was dependent on NPM-ALK, as it significantly decreased after short hairpin RNA (shRNA)–mediated NPM-ALK knock down. In addition, only the constitutively active NPM-ALK, but not the kinase dead NPM-ALKK210R, formed a complex with Shp2, Gab2, and growth factor receptor binding protein 2 (Grb2), where Grb2 bound to the phosphorylated Shp2 through its SH2 domain. Shp2 knock down by specific shRNA decreased the phosphorylation of extracellular signal-regulated kinase 1/2 and of the tyrosine residue Y416 in the activation loop of Src, resulting in impaired ALCL cell proliferation and growth disadvantage. Finally, migration of ALCL cells was reduced by Shp2 shRNA. These findings show a direct involvement of Shp2 in NPM-ALK lymphomagenesis, highlighting its critical role in lymphoma cell proliferation and migration. [Cancer Res 2007;67(9):4278–86]

The use of pumps operating as a turbines (PATs) is an alternative cheap solution for the conversion of dissipations along the distribution networks, but a little information about their performance is available. The turbomachinery affinity law can be applied for the evaluation of the performances curves, but can produce relevant errors that can be reduced with a modification of the affinity law. This research, which is based on experimental collected data, proposes a modification of the turbomachinery affinity law in order to minimize the differences between experimental data and the predicted curves.

Background Treacher Collins syndrome (TCS) is one of the most severe autosomal dominant congenital disorders of craniofacial development and shows variable phenotypic expression. TCS is extremely rare, occurring with an incidence of 1 in 50.000 live births. The TCS distinguishing characteristics are represented by down slanting palpebral fissures, coloboma of the eyelid, micrognathia, microtia and other deformity of the ears, hypoplastic zygomatic arches, and macrostomia. Conductive hearing loss and cleft palate are often present. TCS results from mutations in the TCOF1 gene located on chromosome 5, which encodes a serine/alanine-rich nucleolar phospho-protein called Treacle. However, alterations in the TCOF1 gene have been implicated in only 81-93% of TCS cases. Methods In this study, the entire coding regions of the TCOF1 gene, including newly described exons 6A and 16A, were sequenced in 46 unrelated subjects suspected of TCS clinical indication. Results Fifteen mutations were reported, including twelve novel and three already described in 14 sporadic patients and in 3 familial cases. Moreover, seven novel polymorphisms were also described. Most of the mutations characterised were microdeletions spanning one or more nucleotides, in addition to an insertion of one nucleotide in exon 18 and a stop mutation. The deletions and the insertion described cause a premature termination of translation, resulting in a truncated protein. Conclusion This study confirms that almost all the TCOF1 pathogenic mutations fall in the coding region and lead to an aberrant protein.

Back Cockayne syndrome (CS; MIM 133540-216400) is a rare autosomal recessive neurodegenerative disorder characterized by progressive growth failure, microcephaly, mental retardation, retinal pigmentary degeneration, deafness, photosensitivity, accelerated systemic degeneration of somatic tissue, and premature death. Complementation assays have defined Cockayne syndrome group A (CSA) and Cockayne syndrome group B (CSB), caused by mutations in ERCC8 and ERCC6. The aim of this work was to perform a molecular analysis in a family with an affected son, who died at the age of 12, presenting clinical features typical of CSA. Molecular analysis of ERCC8 allowed us to characterize two novel mutations: a maternally inherited deletion encompassing exons 5 and 6, and a nonsense mutation located in exon 4, segregating from the father. Based on this molecular characterization, we successively performed a prenatal diagnosis on chorionic villus sampling, at 11th week of pregnancy. Molecular prenatal analysis of the ERCC8 was done by analyzing fetal DNA and RNA, looking for both mutations identified in the proband. A linkage analysis was performed using microsatellite markers located on chromosome 5q11 with the purpose to follow the segregation of the mutated alleles within the family. The fetal genotype at CSA locus resulted wild type and was confirmed at birth on biological material isolated from placenta. This study documents for the first time a molecular prenatal diagnosis of CSA, which results in the preferred approach if the mutation within the family is identified in a timely manner.

Sixty-one heterozygotes harboring the germline V804L mutation of the RET protooncogene were identified in five independent families. A total of 31 subjects underwent surgery. Histology identified C cell hyperplasia in 30 cases, isolated in 12 and associated with medullary thyroid carcinoma (MTC) in 18. Six patients with MTC had lymph node metastases. Among the 14 patients with basal detectable calcitonin (CT) level, 12 had MTC and 2 had isolated C cell hyperplasia. In most individuals carrying 804 RET mutation, C cell disease displayed late onset and an indolent course; a pentagastrin test was negative in the majority of heterozygotes during the first 2 decades and was positive in only half of them during the third and fourth decades of life. Interestingly, concomitant somatic M918T was detected in a 12-yr-old girl with MTC and was likely to be responsible for both the early clinical appearance and the aggressiveness of the disease. Our data show that in these gene carriers, surgery may be postponed to the fourth decade of life or until the pentagastrin stimulation test becomes positive. Indeed, our data should be confirmed on a larger series of V804L carriers, but may offer a balanced strategy to keep under control and prevent development of the full disease phenotype.

Background Multinodular goitre (MNG) is a common disorder characterised by an enlargement of the thyroid, occurring as a compensatory response to hormonogenesis impairment. The incidence of MNG is dependent on sex (female:male ratio 5:1) and several reports have documented a genetic basis for the disease. Last year we mapped a MNG locus to chromosome Xp22 in a region containing the peroxiredoxin IV (Prx-IV) gene. Since Prx-IV is involved in the removal of H2O2 in thyroid cells, we hypothesize that mutations in Prx-IV gene are involved in pathogenesis of MNG. Methods Four individuals (2 affected, 2 unrelated unaffected) were sequenced using automated methods. All individuals were originated from the original three-generation Italian family described in previous studies. A Southern blot analysis using a Prx-IV full-length cDNA as a probe was performed in order to exclude genomic rearrangements and/or intronic mutations. In addition a RT-PCR of PRX-IV was performed in order to investigate expression alterations. Results No causative mutations were found. Two adjacent nucleotide substitutions were detected within introns 1 and 4. These changes were also detected in unaffected individuals, suggesting that they were innocuous polymorphisms. No gross genomic rearrangements and/or restriction fragment alterations were observed on Southern analysis. Finally, using RT-PCR from tissue-specific RNA, no differences of PRX-IV expression-levels were detected between affected and unaffected samples. Conclusions Based on sequence and genomic analysis, Prx-IV is very unlikely to be the MNG2 gene.