The normal and fibrotic mouse lung classified by spatial proteomic analysis

Single cell classification is elucidating homeostasis and pathology in tissues and whole organs. We applied in situ spatial proteomics by multiplex antibody staining to routinely processed mouse lung, healthy and during a fibrosis model. With a limited validated antibody panel (24) we classify the normal constituents (alveolar type I and II, bronchial epithelia, endothelial, muscular, stromal and hematopoietic cells) and by quantitative measurements, we show the progress of lung fibrosis over a 4 weeks course, the changing landscape and the cell-specific quantitative variation of a multidrug transporter. An early decline in AT2 alveolar cells and a progressive increase in stromal cells seems at the core of the fibrotic process

Proteomic Fingerprint of Lung Fibrosis Progression and Response to Therapy in Bleomycin-Induced Mouse Model

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by the aberrant accumulation of extracellular matrix in the lungs. nintedanib is one of the two FDA-approved drugs for IPF treatment; however, the exact pathophysiological mechanisms of fibrosis progression and response to therapy are still poorly understood. In this work, the molecular fingerprint of fibrosis progression and response to nintedanib treatment have been investigated by mass spectrometry-based bottom-up proteomics in paraffin-embedded lung tissues from bleomycin-induced (BLM) pulmonary fibrosis mice. Our proteomics results unveiled that (i) samples clustered depending on the tissue fibrotic grade (mild, moderate, and severe) and not on the time course after BLM treatment; (ii) the dysregulation of different pathways involved in fibrosis progression such as the complement coagulation cascades, advanced glycation end products (AGEs) and their receptors (RAGEs) signaling, the extracellular matrix-receptor interaction, the regulation of actin cytoskeleton, and ribosomes; (iii) Coronin 1A (Coro1a) as the protein with the highest correlation when evaluating the progression of fibrosis, with an increased expression from mild to severe fibrosis; and (iv) a total of 10 differentially expressed proteins (padj-value ≤ 0.05 and Fold change ≤-1.5 or ≥1.5), whose abundance varied in the base of the severity of fibrosis (mild and moderate), were modulated by the antifibrotic treatment with nintedanib, reverting their trend. Notably, nintedanib significantly restored lactate dehydrogenase B (Ldhb) expression but not lactate dehydrogenase A (Ldha). Notwithstanding the need for further investigations to validate the roles of both Coro1a and Ldhb, our findings provide an extensive proteomic characterization with a strong relationship with histomorphometric measurements. These results unveil some biological processes in pulmonary fibrosis and drug-mediated fibrosis therapy

BIRTH-WEIGHT OF ITALIAN INFANTS OF 30 WEEKS GESTATION OR LESS

Mean birth weights and percentile charts are given for 161 singleton infants born between 24 and 30 weeks' gestation at the 2nd School of Medicine of Naples. This chart is the first for a Mediterranean population. Our data are similar to those reported from a United Kingdom population and from Japan, suggesting that ethnic differences in birth weight at this gestational age are unimportant

A potential physiological involvement of nesfatin-1 in regulating swine granulosa cell functions

Nesfatin-1 has recently been indicated as a pleiotropic molecule, mainly involved in the metabolic regulation of reproductive functions acting at hypothalamic level. The aim of the present study was to explore a local action of nesfatin-1 in swine ovarian follicle. Firstly, the expression of NUCB-2 by qRT-PCR was verified in swine granulosa cells from different size follicles and nesfatin-1 was localized by immunohistochemistry in sections of the whole porcine ovary. In addition, the in vitro effect of different nesfatin-1 concentrations was tested on cell growth, steroidogenesis and redox status of granulosa cells. Finally, the effect of nesfatin-1 was also studied in an angiogenesis bioassay since vessel growth is essential for ovarian follicle function. Immunohistochemistry revealed intense positivity for nesfatin-1 in swine granulosa cells in all developmental stage follicles. The precursor protein nucleobindin-2 (NUCB-2) showed a higher expression in granulosa cells from large follicles compared to medium and small follicles. In addition, our results demonstrated that nesfatin-1 stimulates cell proliferation and progesterone production and interferes with redox status by modifying NO production and non enzymatic scavenging activity in granulosa cells from large follicles. Moreover, nesfatin-1 displays a stimulatory effects on angiogenesis. Present study demonstrates, for the first time, that nesfatin-1 is physiologically present in the swine ovarian follicle where it could impair granulosa cell functions