Ultrastructural localization of tyrosine hydroxylase in human peripheral blood mononuclear cells: effect of stimulation with phytohaemagglutinin

Using immunocytochemistry coupled to fluorescence and electron microscopy, we investigated the expression and ultrastructural localization of tyrosine hydroxylase (TH, EC 1.14.16.2), the rate-limiting enzyme in the biosynthesis of catecholamines, in human peripheral blood mononuclear cells (PBMCs), with PC12 cells as positive controls. In unstimulated PBMCs, TH-specific immunoreactivity was localized to the plasma membrane. However, after stimulation with the polyclonal mitogen phytohaemagglutinin (PHA), TH immunoreactivity was almost completely localized to electron-dense cytoplasmic granules, which resembled those found in PC12. TH-positive granules, however, were larger (300-500 nm) than in PC12 cells (100-200 nm). Flow cytometry analysis of TH expression showed about 46-50% positive cells in unstimulated PBMCs and in PHA-stimulated PBMCs in the G0/G1 phase of the cell cycle, but more than 80% positive cells in PHA-stimulated PBMCs in the S+G2/M phase. In agreement with previous observations, PHA stimulation also induced de novo expression of TH mRNA as well as increased intracellular catecholamine content, suggesting the occurrence of TH upregulation at the level of both gene expression and enzyme activity. The ultrastructural localization of TH in human PBMCs seems therefore regulated by cell stimulation and related to the functional activity of the enzyme

Studio della quota di volo mediante GNSS, altimetro radar e barometro per rilievi di spettroscopia gamma da velivolo

Lo studio della distribuzione dei radionuclidi terrestri (238U, 232Th e 40K) realizzato mediante tecniche di spettroscopia gamma da velivolo è influenzato dalla quota a cui il rivelatore si trova rispetto al suolo. Un'incertezza del 10% a 100 m di altezza origina un errore nella stima del segnale gamma del 208Tl, appartenente alla catena di decadimenti del 232Th, dell’ordine del 7%. L'impiego di una nuova classe di spettrometri montati a bordo di UAV (Unmanned Aerial Vehicle) per raffinate misure in contesti ostili o remoti rende necessaria un'accurata stima in real time della quota di volo. Il Radgyro è un velivolo dedicato a survey multiparametrici, capace di trasportare strumentazione pari ad un payload massimo di 120 kg, tra cui quattro spettrometri gamma NaI(Tl). Una stazione inerziale con ricevitore integrato GNSS (Global Navigation Satellite System) restituisce l'assetto del velivolo con una frequenza massima di 400 Hz. Il velivolo è dotato di un network di tre ricevitori GNSS posizionati alle estremità della carena del velivolo. Un altimetro radar a 24 GHz rileva la quota con una frequenza di 60 Hz. La misura di pressione e temperatura consente di ricavare la quota barometrica a 2 Hz. Con l'obiettivo di studiare le incertezze associate alle misure della quota di volo acquisite dagli altimetri in relazione ai dati GNSS, sono stati realizzati tre voli sul mare in un range di altezze comprese tra 31 m e 249 m, per un totale di 4702 secondi di volo effettivo. Al termine dello studio è possibile concludere che l'errore complessivo delle abbondanze di K, U e Th aumenta di 7.7%, 0.5% e 2.7% rispettivamente, a causa delle incertezze della quota di volo

Growth and mechanical correlations of calcified cartilage in Batoidea: A histomorphological study using the Raja asterias model

This study investigates the growth and calcification of the appendicular skeleton in Raja asterias (Delaroche, 1809), a member of the Batoidea, to explore the relationship between histomorphology and the mechanics of batoid locomotion within the water column. Although much prior research has focused on the "tessellated pattern" in these fishes, the variable structure of the appendicular skeleton provides fresh insights into the understudied interplay between skeletal histomorphology and the mechanical functions of Batoidea fins. The shape and initial growth of fin cartilage are influenced by the orientation of chondrocyte mitoses prior to mineral deposition, with subsequent calcification playing a pivotal role in shaping skeletal architecture. This study documents two distinct growth patterns: "crustal" and "catenated." The crustal pattern is predominantly observed in larger skeletal elements, such as the central body structures (skull, rostrum, and jaws), girdles, pterygia, and compound radials, whereas fin radials follow the catenated growth pattern. Notably, early-stage chondrichthyan cartilage shares similarities with mammalian metaphyseal growth plate cartilage, though in chondrichthyans, the calcified matrix is not resorbed or replaced by bone. Additionally, a previously unrecognized calcification pattern is identified in the pelvic-fin radials of R. asterias, indicating that the mechanical demands of locomotion in the water column may have driven the evolution of variable fin flexibility in Batoidea. This flexibility is achieved through joint mobility (diarthroses and amphiarthroses), specialized fin structures, and the distinct calcification patterns of the pectoral and pelvic fins

Comparative morphology of skeletal development in homo sapiens and raja asterias: divergent stiffening patterns due to different matrix calcification processes.

Before calcification begins, the early embryonic and fetal skeletal development of both mammalian Homo sapiens and the chondrichthyan fish Raja asterias consists exclusively of cartilage. This cartilage is formed and shaped through processes involving tissue segmentation and the frequency, distribution, and orientation of chondrocyte mitoses. In the subsequent developmental phase, mineral deposition in the cartilage matrix conditions the development further. The stiffness and structural layout of the mineralized cartilage have a significant impact on the shape of the anlagen (early formative structure of a tissue, a scaffold on which the new bone is formed) and the mechanical properties of the skeletal segments. The fundamental difference between the two studied species lies in how calcified cartilage serves as a scaffold for osteoblasts to deposit bone matrix, which is then remodeled. In contrast, chondrichthyans retain the calcified cartilage as the definitive skeletal structure. This study documents the distinct mineral deposition pattern in the cartilage of the chondrichthyan R. asterias, in which calcification progresses with the formation of focal calcification nuclei or “tesserae”. These are arranged on the flat surface of the endo-skeleton (crustal pattern) or aligned in columns (catenated pattern) in the radials of the appendicular skeleton. This anatomical structure is well adapted to meet the mechanical requirements of locomotion in the water column. Conversely, in terrestrial mammals, endochondral ossification (associated with the remodeling of the calcified matrix) provides limb bones with the necessary stiffness to withstand the strong bending and twisting stresses of terrestrial locomotion. In this study, radiographs of marine mammals (reproduced from previously published studies) document how the endochondral ossification in dolphin flippers adapts to the mechanical demands of aquatic locomotion. This adaptation includes the reduction in the length of the stylopodium and zeugopodium and an increase in the number of elements in the autopodium’s central rays

The combined cartilage growth – calcification patterns in the wing-fins of Rajidae (Chondrichthyes): A divergent model from endochondral ossification of tetrapods

The relationship between cartilage growth – mineralization patterns were studied in adult Rajidae with X-ray morphology/morphometry, undecalcified resin-embedded, heat-deproteinated histology and scanning electron microscopy. Morphometry of the wing-fins, nine central rays of the youngest and oldest specimens documented a significant decrement of radials mean length between inner, middle and outer zones, but without a regular progression along the ray. This suggests that single radial length growth is regulated in such a way to align inter-radial joints parallel to the wing metapterygia curvature. Trans-illumination and heat-deproteination techniques showed polygonal and cylindrical morphotypes of tesserae, whose aligned pattern ranged from mono-columnar, bi-columnar, and multi-columnar up to the crustal-like layout. Histology of tessellated cartilage allowed to identify of zones of the incoming mineral deposition characterized by enhanced duplication rate of chondrocytes with the formation of isogenic groups, whose morphology and topography suggested a relationship with the impending formation of the radials calcified column. The morphotype and layout of radial tesserae were related to mechanical demands (stiffening) and the size/mass of the radial cartilage body. The cartilage calcification pattern of the batoids model shares several morphological features with tetrapods' endochondral ossification, that is, (chondrocytes' high duplication rate, alignment in rows, increased volume of chondrocyte lacunae), but without the typical geometry of the metaphyseal growth plates

The collagenic architecture of human dura mater: Laboratory investigation

Object. Human dura mater is the most external meningeal sheet surrounding the CNS. It provides an efficient protection to intracranial structures and represents the most important site for CSF turnover. Its intrinsic architecture is made up of fibrous tissue including collagenic and elastic fibers that guarantee the maintenance of its biophysical features. The recent technical advances in the repair of dural defects have allowed for the creation of many synthetic and biological grafts. However, no detailed studies on the 3D microscopic disposition of collagenic fibers in dura mater are available. The authors report on the collagenic 3D architecture of normal dura mater highlighting the orientation, disposition in 3 dimensions, and shape of the collagen fibers with respect to the observed layer. Methods. Thirty-two dura mater specimens were collected during cranial decompressive surgical procedures, fixed in 2.5% Karnovsky solution, and digested in 1 N NaOH solution. After a routine procedure, the specimens were observed using a scanning electron microscope. Results. The authors distinguished the following 5 layers in the fibrous dura mater of varying thicknesses, orientation, and structures: bone surface, external median, vascular, internal median, and arachnoid layers. Conclusions. The description of the ultrastructural 3D organization of the different layers of dura mater will give us more information for the creation of synthetic grafts that are as similar as possible to normal dura mater. This description will be also related to the study of the neoplastic invasion

Metagenome-sourced microbial chitinases as potential insecticide proteins

Microbial chitinases are gaining interest as promising candidates for controlling plant pests. These enzymes can be used directly as biocontrol agents as well as in combination with chemical pesticides or other biopesticides, reducing their environmental impact and/or enhancing their efficacy. Chitinolytic enzymes can target two different structures in insects: the cuticle and the peritrophic matrix (PM). PM, formed by chitin fibrils connected to glycoproteins and proteoglycans, represents a physical barrier that plays an essential role in midgut physiology and insect digestion, and protects the absorptive midgut epithelium from food abrasion or pathogen infections. In this paper, we investigate how two recently discovered metagenomesourced chitinases (Chi18H8 and 53D1) affect, in vitro and in vivo, the PM integrity of Bombyx mori, a model system among Lepidoptera. The two chitinases were produced in Escherichia coli or, alternatively, in the unconventional \u2013 but more environmentally acceptable \u2013 Streptomyces coelicolor. Although both the proteins dramatically altered the structure of B. mori PM in vitro, when administered orally only 53D1 caused adverse and marked effects on larval growth and development, inducing mortality and reducing pupal weight. These in vivo results demonstrate that 53D1 is a promising candidate as insecticide protein

Il Radgyro: un autogiro dedicato ad acquisizioni airborne multiparametriche

This study summarizes the main characteristic of the Radgyro, an auotgyro dedicated to multiparametric airborne acquisitions. This aircraft was imagined, designed and realized to host two cameras, an infrared camera, a thermographic camera, five gamma-ray spectrometers as well as a GPS antenna network, a inertial navigation system, a radar altimeter and the electronic system necessary to the automatic, simultaneous and independent use of the entire instrumentation. Thanks to a carrying capacity of 130 kg, 4 hours of flight autonomy and a low overhead, the Radgyro is the ideal vehicle for multiparametric surveys of wide areas, where the use of drones is limited by the carrying capacity and by the flight autonomy. Finally the results of preliminary photogrammetric surveys and the possible future applications of the aircraft are presented