La vaghezza dell’oggetto di diagnosi e la proliferazione del linguaggio diagnostico. Sviluppo storico

A partire dal Settecento la malattia viene concettualmente precisata in quanto “lesione locale”; parallelamente nasce e si diffonde la nosologia, che include nomi di malattie e di varie situazioni esistenziali. Verso la fine del Novecento si assiste a una forte proliferazione del linguaggio diagnostico. L’articolo ne dà una prima sistemazione teorica. Historical research shows that the concept of disease, i.e. the prototypical object of diagnosis, has always been rather vague, and only during the 18th and 19th centuries it acquired a more specific connotation as “local lesion”. The same period saw the development of nosology, which, in addition to the names of the various diseases, also included terms referring to different existential conditions, feelings and behaviours, thus encouraging the further establishment of an ambiguous concept of object of diagnosis. Starting from the end of the 19th century an international medical language began to spread, while during the 20th century the concept of “local lesion” no longer appeared as a prerequisite for the definition of a disease. 

MENINGKATKAN KEMAMPUAN BERPIKIR KRITIS IPA MELALUI MODEL PEMBELAJARAN Children Learning In Science (CLIS) PADA SISWA KELAS IV SDN MENTENG ATAS 06 PAGI JAKARTA SELATAN

Penelitian tindakan kelas ini bertujuan mengetahui penerapan model pembelajaranChildren Learning In Science (CLIS) terhadap kemampuan berpikir kritis IPA pada siswa kelas IV SDN Menteng Atas 06 Pagi Jakarta Selatan. Penelitian ini dilaksanakn di SDN Menteng Atas 06 Pagi Jakarta Selatan pada bulan Januari 2017, dengan subjek penelitian siswa kelas IV yang berjumlah 27 siswa. Model penelitian yang digunakan adalah penelitian tindakan kelas dari Kemmis dan Taggart dengan empat tahap yaitu, tahap perencanaan, pelaksanaan, tindakan, pengamatan observasi dan refleksi. Penelitian ini dilakukan sebanyak dua siklus. Penelitian dilakukan dengan menggunakan lembar observasi aktivitas guru, siswa, instrumen tes avaluasi berupa essy dan dokumen aktivitas pembelajaran. Berdasarkan data dan hasil kemampuan berpikir krtitis siswa dalam menyelesaikan tes evaluasi dan beberapa percobaan menunjukan bahwa hasil pada siklus I diperoleh nilai ketuntasan sebesar 55,00% siklus II mencapai 88,89% dari jumlah siswa. Adapun aktivitas siswa siklus I adala 76,66% dan siklus II mencapai 90%. Maka dikatakn bahwa jumlah siswa yang mencapai target sebanyak 20 siswa mencapai target skor kemampuan berpikir kritis IPA ≥ 70. Berdasarkan hasil penelitian ini dapat disimpulkan bahwa penerapan model pembelajaraan Children Learning In Science (CLIS) pada pembelajaran IPA tentang Energi dapat meningkatkan kemampuan berpikir kritis siswa dari tes tertulis berupa tes essy dan percobaan. Keaktivitas siswa dan kualitas siswa mengikuti pembelajaran di kelas SDN Menteng Atas 06 Pagi Jakarta Selatan, implikasi dari hasil penelitian adalah model pembelajaran Children Learning In Science (CLIS) dapat dijadikan sebagai salah satu model belajar untuk mempermudahkan siswa dalam memahami mata pelajaran IPA melalui materi Energi yang berkaitan dengan kehidupan sehari-hari. This classroom action research aims to determine the application of the model learning Children Learning In Science (CLIS) on the ability of critical thinking IPA in grade IV SDN 06 Menteng Morning South Jakarta. This study held at SDN 06 Menteng top Morning South Jakarta in January 2017, with research subjects fourth graders totaling 27 students. Model of research is classroom action research of Kemmis and Taggart with four stages, namely, planning, implementation, action, observation and reflection observation. This study was conducted by two cycles. The study was conducted by using observation sheet activities of teachers, students, test instruments essay evaluation form and document learning activities. Based on the data and the results of students' ability to think critical in completing the evaluation tests and several experiments show that the results obtained in the first cycle completeness value of 55.00% cycle II reached 88.89% of the total number of students. The student activity is 76.66% the first cycle and the second cycle was 90%. So said that the number of students who achieve a target of 20 students reached the target score of critical thinking skills IPA ≥ 70. Based on these results it can be concluded that the application of the learning model Children Learning In Science (CLIS) in science teaching on Energy can improve students' critical thinking skills of essy written test is test and trial. Creativity students and the quality of students attend classroom learning SDN Menteng top 06 Morning South Jakarta, the implications of the study are learning models Children Learning In Science (CLIS) could serve as a model to learn to facilitate students in understanding science subjects through the material energy related to everyday life

All about alles: The syntax of wh-quantifier float in German

This thesis offers an in-depth investigation of “wh-quantifier float” of the quantifying particle ‘alles’ in German. 'Alles' (etymologically, ‘all’) appears in wh-questions like 'Wen alles hat die Mare eingeladen?' (‘Who-all did Mare invite?’). The thesis focuses on the syntactic distribution of 'alles'. 'Alles' enjoys a wide distribution in the clause. It can occur both ‘adjacent’ to its ‘associate’ wh-phrase, and ‘distant’ from it, in various positions of the clause. I address three questions: What determines the distribution of 'alles'? Are adjacent 'alles' and ‘distal alles’ the same category? What licenses distal 'alles'? I answer these questions by arguing for a stranding analysis of distal 'alles': 'alles' and its associate form a first-Merge constituent, which is optionally separated in the course of the derivation through a process that involves movement ([WH alles] ⇒ [WH. . . [[WH alles]. . . ]]). The conclusion is compatible with prior analyses that argued for or assumed (a) constituency, and (b) a movement dependency in overt syntax. The conclusion is at odds with adverbial analyses, which assume that distal 'alles' is an adverbial. I provide two main empirical arguments. First, I argue against the idea that distal 'alles' and adjacent 'alles' are separate lexical items, or have different lexical content. Second, I argue that the “Chain Link Generalization” is the most accurate generalization for the distribution of 'alles': Given a derivation involving 'alles' and a licit associate, 'alles' may appear in any position which hosts an Abar-chain link of the associate, and in no other position. I show that 'alles' has “no distribution of its own in the clause”. Rather, the distribution of 'alles' depends on the potential distribution of its associate and can be predicted by the associate’s category, the associate’s base-position, the derivation that the associate undergoes in a given sentence. Conceptually, I argue that a stranding analysis is favored by simplicity as most generalizations established in this dissertation are directly entailed by it

Avian neural crest cell migration is diversely regulated by the two major hyaluronan-binding proteoglycans PG-M/versican and aggrecan

It has been proposed that hyaluronan-binding proteoglycans play an important role as guiding cues during neural crest (NC) cell migration, but their precise function has not been elucidated. In this study, we examine the distribution, structure and putative role of the two major hyaluronan-binding proteoglycans, PG-M/versicans and aggrecan, during the course of avian NC development. PG-M/versicans V0 and V1 are shown to be the prevalent isoforms at initial and advanced phases of NC cell movement, whereas the V2 and V3 transcripts are first detected following gangliogenesis. During NC cell dispersion, mRNAs for PG-M/versicans V0/V1 are transcribed by tissues lining the NC migratory pathways, as well as by tissues delimiting nonpermissive areas. Immunohistochemistry confirm the deposition of the macromolecules in these regions and highlight regional differences in the density of these proteoglycans. PG-M/versicans assembled within the sclerotome rearrange from an initially uniform distribution to a preferentially caudal localization, both at the mRNA and protein level. This reorganization is a direct consequence of the metameric NC cell migration through the rostral portion of the somites. As suggested by previous in situ hybridizations, aggrecan shows a virtually opposite distribution to PG-M/versicans being confined to the perinotochordal ECM and extending dorsolaterally in a segmentally organized manner eventually to the entire spinal cord at axial levels interspacing the ganglia. PG-M/versicans purified from the NC migratory routes are highly polydispersed, have an apparent M(r) of 1,200-2,000 kDa, are primarily substituted with chondroitin-6-sulfates and, upon chondroitinase ABC digestion, are found to be composed of core proteins with apparent M(r)of 360–530, 000. TEM/rotary shadowing analysis of the isolated PG-M/versicans confirmed that they exhibit the characteristic bi-globular shape, have core proteins with sizes predicted for the V0/V1 isoforms and carry relatively few extended glycosaminoglycan chains. Orthotopical implantation of PG-M/versicans immobilized onto transplantable micromembranes tend to ‘attract’ moving cells toward them, whereas similar implantations of a notochordal type-aggrecan retain both single and cohorts of moving NC cells in close proximity of the implant and thereby perturb their spatiotemporal migratory pattern. NC cells fail to migrate through three-dimensional collagen type I-aggrecan substrata in vitro, but locomote in a haptotactic manner through collagen type I-PG-M/versican V0 substrata via engagement of HNK-1 antigen-bearing cell surface components. The present data suggest that PG-M/versicans and notochordal aggrecan exert divergent guiding functions during NC cell dispersion, which are mediated by both their core proteins and glycosaminoglycan side chains and may involve ‘haptotactic-like’ motility phenomena. Whereas aggrecan defines strictly impenetrable embryonic areas, PG-M/versicans are central components of the NC migratory pathways favoring the directed movement of the cells

Isolation and Characterization of EMILIN-2, a New Component of the Growing EMILINs Family and a Member of the EMI Domain-containing Superfamily

EMILIN (elastin microfibril interfase located Protein) is an elastic fiber-associated glycoprotein consisting of a self-interacting globular C1q domain at the C terminus, a short collagenous stalk, an extended region of potential coiled-coil structure, and an N-terminal cysteine-rich domain (EMI domain). Using the globular C1q domain as a bait in the yeast two-hybrid system, we have isolated a cDNA encoding a novel protein. Determination of the entire primary structure demonstrated that this EMILIN-binding polypeptide is highly homologous to EMILIN. The domain organization is superimposable, one important difference being a proline-rich (41%) segment of 56 residues between the potential coiled-coil region and the collagenous domain absent in EMILIN. The entire gene (localized on chromosome 18p11.3) was isolated from a BAC clone, and it is structurally almost identical to that of EMILIN (8 exons, 7 introns with identical phases at the exon/intron boundaries) but much larger (about 40 versus 8 kilobases) than that of EMILIN. Given these findings we propose to name the novel protein EMILIN-2 and the prototype member of this family EMILIN-1 (formerly EMILIN). The mRNA expression of EMILIN-2 is more restricted compared with that of EMILIN-1; highest levels are present in fetal heart and adult lung, whereas, differently from EMILIN-1, adult aorta, small intestine, and appendix show very low expression, and adult uterus and fetal kidney are negative. Finally, the EMILIN-2 protein is secreted extracellularly by in vitro-grown cells, and in accordance with the partial coexpression in fetal and adult tissues, the two proteins shown extensive but not absolute immunocolocalization in vitro

Structure, Chromosomal Localization, and Promoter Analysis of the Human Elastin MicrofibrilInterfase Located proteIN (EMILIN) Gene

Abstract Elastinmicrofibril interfase-located protein (EMILIN) is an extracellular matrix glycoprotein abundantly expressed in elastin-rich tissues such as the blood vessels, skin, heart, and lung. It occurs with elastic fibers at the interface between amorphous elastin and microfibrils. In vitroexperiments suggested a role for EMILIN in the process of elastin deposition. This multimodular protein consists of 995 amino acids; the domain organization includes a C1q-like globular domain at the C terminus, a short collagenous stalk, a region containing two leucine zippers, and at least four heptad repeats with a high potential for forming coiled-coil α-helices and, at the N terminus, a cysteine-rich sequence characterized by a partial epidermal growth factor-like motif and homologous to a region of multimerin. Here we report the complete characterization of the human and murine EMILIN gene, their chromosomal assignment, and preliminary functional data of the human promoter. A cDNA probe corresponding to the C terminus of EMILIN was used to isolate two genomic clones from a human BAC library. Sequencing of several derived subclones allowed the characterization of the whole gene that was found to be about 8 kilobases in size and to contain 8 exons and 7 introns. The internal exons range in size from 17 base pairs to 1929 base pairs. All internal intron/exon junctions are defined by canonical splice donor and acceptor sites, and the different domains potentially involved in the formation of a coiled-coil structure are clustered in the largest exon. The 3′-end of the EMILIN gene overlaps with the 5′-end of the promoter region of the ketohexokinase gene, whose chromosomal position is between markers D2S305 and D2S165 on chromosome 2. A 1600-base pair-long sequence upstream of the translation starting point was evaluated for its promoter activity; five deletion constructs were assayed after transfection in primary chicken fibroblasts and in a human rhabdomyosarcoma cell line. This analysis indicates the existence of two contiguous regions able to modulate luciferase expression in both cell types used, one with a strong activatory function, ranging from positions −204 to −503, and the other, ranging from positions −504 to −683, with a strong inhibitory function

Multiplex staining depicts the immune infiltrate in colitis-induced colon cancer model

Assessment of the host immune response pattern is of increasing importance as highly prognostic and diagnostic, in immune-related diseases and in some types of cancer. Chronic inflammation is a major hallmark in colon cancer formation, but, despite the extent of local inflammatory infiltrate has been demonstrated to be extremely informative, its evaluation is not routinely assessed due to the complexity and limitations of classical immunohistochemistry (IHC). In the last years, technological advance helped in bypassing technical limits, setting up multiplex IHC (mIHC) based on tyramide signal amplification (TSA) method and designing software suited to aid pathologists in cell scoring analysis. Several studies verified the efficacy of this method, but they were restricted to the analysis of human samples. In the era of translational medicine the use of animal models to depict human pathologies, in a more complete and complex approach, is really crucial. Nevertheless, the optimization and validation of this method to species other than human is still poor. We took advantage of Multispectral Imaging System to identify the immunoprofile of Dextran Sulphate Sodium (DSS)-treated mouse colon. We optimized a protocol to sequentially stain formalin fixed paraffin embedded murine colon samples for CD3, CD8a, CD4, and CD4R5B0 antigens. With this approach we obtained a detailed lymphocyte profile, while preserving the morphological tissue context, generally lost with techniques like gene expression profiling or flow cytometry. This study, comparing the results obtained by mIHC with immunophenotyping performed with cytofluorimetric and standard IHC methods validates the potentiality and the applicability of this innovative approach

Role of extracellular matrix in gastrointestinal cancer-associated angiogenesis

Gastrointestinal tumors are responsible for more cancer-related fatalities than any other type of tumors, and colorectal and gastric malignancies account for a large part of these diseases. Thus, there is an urgent need to develop new therapeutic approaches to improve the patients\u2019 outcome and the tumor microenvironment is a promising arena for the development of such treatments. In fact, the nature of the microenvironment in the different gastrointestinal tracts may significantly influence not only tumor development but also the therapy response. In particular, an important microenvironmental component and a potential therapeutic target is the vasculature. In this context, the extracellular matrix is a key component exerting an active effect in all the hallmarks of cancer, including angiogenesis. Here, we summarized the current knowledge on the role of extracellular matrix in affecting endothelial cell function and intratumoral vascularization in the context of colorectal and gastric cancer. The extracellular matrix acts both directly on endothelial cells and indirectly through its remodeling and the consequent release of growth factors. We envision that a deeper understanding of the role of extracellular matrix and of its remodeling during cancer progression is of chief importance for the development of new, more efficacious, targeted therapies

Integrin binding site within the gC1q domain orchestrates EMILIN-1-induced lymphangiogenesis.

Lymphatic vessels (LVs) play a pivotal role in the control of tissue homeostasis and also have emerged as important regulators of immunity, inflammation and tumor metastasis. EMILIN-1 is the first ECM protein identified as a structural modulator of the growth and maintenance of LV; accordingly, Emilin1-/- mice display lymphatic morphological alterations leading to functional defects as mild lymphedema, leakage and compromised lymph drainage. Many EMILIN-1 functions are exerted by the binding of its gC1q domain with the E933 residue of α4 and α9β1 integrins. To investigate the specific regulatory role of this domain on lymphangiogenesis, we generated a transgenic mouse model expressing an E933A-mutated EMILIN-1 (E1-E933A), unable to interact with α4 or α9 integrin. The mutant resulted in abnormal LV architecture with dense, tortuous and irregular networks; moreover, the number of anchoring filaments was reduced and collector valves had aberrant narrowed structures. E933A mutation also affected lymphatic function in lymphangiography assays and made the transgenic mice more prone to lymph node metastases. The finding that the gC1q/integrin interaction is crucial for a correct lymphangiogenesis response was confirmed and reinforced by functional in vitro tubulogenesis assays. In addition, ex vivo thoracic-duct ring assays revealed that E1-E933A-derived lymphatic endothelial cells had a severe reduction in sprouting capacity and were unable to organize into capillary-like structures. All these data provide evidence that the novel "regulatory structural" role of EMILIN-1 in the lymphangiogenic process is played by the integrin binding site within its gC1q domain

Engineering novel complement activity into a pulmonary surfactant protein

Complement neutralizes invading pathogens, stimulates inflammatory and adaptive immune responses, and targets non- or altered-self structures for clearance. In the classical and lectin activation pathways, it is initiated when complexes composed of separate recognition and activation subcomponents bind to a pathogen surface. Despite its apparent complexity, recognition-mediated activation has evolved independently in three separate protein families, C1q, mannose-binding lectins (MBLs), and serum ficolins. Although unrelated, all have bouquet-like architectures and associate with complement-specific serine proteases: MBLs and ficolins with MBL-associated serine protease-2 (MASP-2) and C1q with C1r and C1s. To examine the structural requirements for complement activation, we have created a number of novel recombinant rat MBLs in which the position and orientation of the MASP-binding sites have been changed. We have also engineered MASP binding into a pulmonary surfactant protein (SP-A), which has the same domain structure and architecture as MBL but lacks any intrinsic complement activity. The data reveal that complement activity is remarkably tolerant to changes in the size and orientation of the collagenous stalks of MBL, implying considerable rotational and conformational flexibility in unbound MBL. Furthermore, novel complement activity is introduced concurrently with MASP binding in SP-A but is uncontrolled and occurs even in the absence of a carbohydrate target. Thus, the active rather than the zymogen state is default in lectin·MASP complexes and must be inhibited through additional regions in circulating MBLs until triggered by pathogen recognition