The behaviour of osteocyte dendritic processes in bone under cyclic load (il comportamento dei processi dendritici degli osteociti nel tessuto osseo sottoposti a carico ciclico)

Bone is continually being removed and replaced through the actions of basic multicellular units (BMU). This constant upkeep is necessary to remove microdamage formed naturally due to fatigue and thus maintain the integrity of the bone. The repair process in bone is targeted, meaning that a BMU travels directly to the site of damage and repairs it. It is still unclear how targeted remodelling is stimulated and directed but it is highly likely that osteocytes play a role. A number of theories have been advanced to explain the microcrack osteocyte interaction but no complete mechanism has been demonstrated. Osteocytes are connected to each other by dendritic processes. The “scissors model" proposed that the rupture of these processes where they cross microcracks signals the degree of damage and the urgency of the necessary repair. In its original form it was proposed that under applied compressive loading, microcrack faces will be pressed together and undergo relative shear movement. If this movement is greater than the width of an osteocyte process, then the process will be cut in a “scissors like" motion, releasing RANKL, a cytokine known to be essential in the formation of osteoclasts from pre-osteoclasts. The main aim of this thesis was to investigate this theoretical model with a specific focus on microscopy and finite element modelling. Previous studies had proved that cyclic stress was necessary for osteocyte process rupture to occur. This was a divergence from the original “scissors model" which had proposed that the cutting of cell material occurred in one single action. The present thesis is the first study to show fatigue failure in cellular processes spanning naturally occurring cracks and it's the first study to estimate the cyclic strain range and relate it to the number of cycles to failure, for any type of cell. Rupture due to shear movement was ruled out as microcrack closing never occurred, as a result of plastic deformation of the bone. Fatigue failure was found to occur due to cyclic tensile stress in the locality of the damage. The strain range necessary for osteocyte process rupture was quantified. It was found that the lower the process strain range the greater the number of cycles to cell process failure. FEM modelling allowed to predict stress in the vicinity of an osteocyte process and to analyse its interaction with the bone surrounding it: simulations revealed evident creep effects in bone during cyclic loading. This thesis confirms and dismisses aspects of the “scissors model". The observations support the model as a viable mechanism of microcrack detection by the osteocyte network, albeit in a slightly modified form where cyclic loading is necessary and the method of rupture is fatigue failure due to cyclic tensile motion. An in depth study was performed focusing on microscopy analysis of naturally occurring cracks in bone and FEM simulation analysis of an osteocyte process spanning a microcrack in bone under cyclic load.

Safety factors in sheet pile wall design: considerations by using traditional methods and FEM analysis

The purpose of the work is: define and calculate a factor of collapse related to traditional method to design sheet pile walls. Furthermore, we tried to find the parameters that most influence a finite element model representative of this problem. The text is structured in this way: from chapter 1 to 5, we analyzed a series of arguments which are usefull to understanding the problem, while the considerations mainly related to the purpose of the text are reported in the chapters from 6 to 10. In the first part of the document the following arguments are shown: what is a sheet pile wall, what are the codes to be followed for the design of these structures and what they say, how can be formulated a mathematical model of the soil, some fundamentals of finite element analysis, and finally, what are the traditional methods that support the design of sheet pile walls. In the chapter 6 we performed a parametric analysis, giving an answer to the second part of the purpose of the work. Comparing the results from a laboratory test for a cantilever sheet pile wall in a sandy soil, with those provided by a finite element model of the same problem, we concluded that:in modelling a sandy soil we should pay attention to the value of cohesion that we insert in the model (some programs, like Abaqus, don’t accept a null value for this parameter), friction angle and elastic modulus of the soil, they influence significantly the behavior of the system (structure-soil), others parameters, like the dilatancy angle or the Poisson’s ratio, they don’t seem influence it. The logical path that we followed in the second part of the text is reported here. We analyzed two different structures, the first is able to support an excavation of 4 m, while the second an excavation of 7 m. Both structures are first designed by using the traditional method, then these structures are implemented in a finite element program (Abaqus), and they are pushed to collapse by decreasing the friction angle of the soil. The factor of collapse is the ratio between tangents of the initial friction angle and of the friction angle at collapse. At the end, we performed a more detailed analysis of the first structure, observing that, the value of the factor of collapse is influenced by a wide range of parameters including: the value of the coefficients assumed in the traditional method and by the relative stiffness of the structure-soil system. In the majority of cases, we found that the value of the factor of collapse is between and 1.25 and 2. With some considerations, reported in the text, we can compare the values so far found, with the value of the safety factor proposed by the code (linked to the friction angle of the soil).

Stock boundaries of Parapenaeus longirostris (Lucas, 1846) populations in the Italian seas: Development of genomic 2b-RAD markers and biological variation analysis.

This thesis is developed in the contest of Ritmare project WP1, which main objective is the development of a sustainable fishery through the identification of populations boundaries in commercially important species in Italian Seas. Three main objectives are discussed in order to help reach the main purpose of identification of stock boundaries in Parapenaeus longirostris: 1 -Development of a representative sampling design for Italian seas; 2 -Evaluation of 2b-RAD protocol; 3 -Investigation of populations through biological data analysis. First of all we defined and accomplished a sampling design which properly represents all Italian seas. Then we used information and data about nursery areas distribution, abundance of populations and importance of P. longirostris in local fishery, to develop an experimental design that prioritize the most important areas to maximize the results with actual project funds. We introduced for the first time the use of 2b-RAD on this species, a genotyping method based on sequencing the uniform fragments produced by type IIB restriction endonucleases. Thanks to this method we were able to move from genetics to the more complex genomics. In order to proceed with 2b-RAD we performed several tests to identify the best DNA extraction kit and protocol and finally we were able to extract 192 high quality DNA extracts ready to be processed. We tested 2b-RAD with five samples and after high-throughput sequencing of libraries we used the software “Stacks” to analyze the sequences. We obtained positive results identifying a great number of SNP markers among the five samples. To guarantee a multidisciplinary approach we used the biological data associated to the collected samples to investigate differences between geographical samples. Such approach assures continuity with other project, for instance STOCKMED, which utilize a combination of molecular and biological analysis as well.

Double Dyke System - Natural engineering solutions

According to various studies, the effects of climate change will be a danger to ecosystems and the population, especially in coastal areas, increasing the risk of floods. Authorities are taking action to prevent future disasters using traditional engineering solutions. These solutions can have high environmental and economic costs, fixing the coastline, increasing the salinization of aquifers, and can be subject to failure mechanisms. For this reason, studies were made to use natural engineering solutions for coastal protection, instead of traditional solutions, to achieve the UN SDGs. Coastal ecosystems have the natural ability to repair and restore themselves, increasing soil elevation, and attenuating waves. One of these solutions is the Double Dyke System, consisting of creating a salt marsh between the first dyke and a second inland. The goal is to protect the coasts and to restore ecosystems. The purpose of this study is to compare the costs of natural engineering solutions with traditional ones. It is assumed that these solutions may be more effective and less expensive in the long run. For this evaluation, a suitability analysis of the polders in the Dutch Zeeland region to assess the costs and benefits under different SLR scenarios was made. A saline intrusion model was also created to analyze the effects of a salt marsh on the aquifers. From the analyzes conducted, the implementation of the DDS turns out to be the cheapest coastal defense system in all SLR scenarios. The presence of a salt marsh could also have a positive impact on the prevention of saline intrusion in the various scenarios considered. The DDS could have a positive economic and environmental impact in the long term, reducing the investment costs for coastal defense and bringing important benefits for the protection of man and nature. Despite the results, more studies are needed on the efficiency of this defense system and on the economic evaluation of non-marketable ecosystem services.

Steel hopper silos filled with granular material: experimental tests on the tensile resistance of corrugated sheets and on the filling and discharging pressures

This thesis addresses various aspects related to silos, from the strength of some structural parts to internal actions due to grain. Two hopper silo models were mainly studied, so the thesis is divided into two parts. The first part focuses only on the silo cylinder and deals with the collapse of a silo due to failure of the vertical walls. We had the opportunity to access data from a real silo and perform tensile tests on corrugated sheets. The theoretical and experimental resistance of the corrugated sheet forming the silo cylinder was studied. The resistance was then compared with the internal actions due to grain prescribed by various standards. The second part, however, focused on the hopper of a silo in which a load test (loading and unloading of the silo) was performed. Through the test data, an attempt was made to reproduce the pressures normal to the hopper through analytical reasoning. The experimental pressures were then compared with the theoretical pressures predicted by the standards. In addition, with mathematical reasoning, an attempt was made to reproduce the horizontal pressure on the vertical walls of the silo from the experimental normal pressure in the hopper. In fact, the test was related only to the hopper part and not to the silo cylinder.

Determination of microplastics in the digestive content of pelagic fish of commercial interest

The study aimed to determine the amount of microplastics (MPs) found in the digestive content of some pelagic tuna fish species of commercial interest caught along the Spanish coasts. In total, 601 individuals belonging to eight tuna species, and one species of the Corypheanidae family, were examined for the presence of MPs in their stomachs. Fish were collected from 9 different locations along the Spanish coasts by commercial fishers. A total of 170 MPs were extracted from the fish stomach of 75 individuals. The number of particles present in the stomach ranged from 1 to 20 (mean 0.3± 1.2 SD MPs per individual). The species with the highest amount of MPs was Auxix rochei whereas the species that did not present any MPs was Coryphaena hippurus, the only species not belonging to the tuna group. In terms of location, the highest ratio of MPs (71%) was found in the fish collected from Azohia, and the lowest in Mazagon (7%). In general, the fish that ingested more MPs were those caught from sites that were closer to the coasts (Canarias Islands stations and Azohia). No correlation between the number of ingested MPs and the size and weight of the fish were detected nor for the whole sample nor for each species separately. Analyzing the MPs in terms of colour, green colour MPs were the most dominant (52%) followed by blue colour (25%). Analyzing the MPs in terms of size, 11 categories were considered and the most dominant size was 0-0.5 micron. Fibers were excluded from the analysis because no protocol was adopted to prevent their contamination. Furthermore, 100% of the MPs found were fragments. The data did not show a high presence of MPs in the analyzed study areas. A bias could probably be due to the method used, allowing to loose a certain quantity of very small microplastics, and also to ethological and biological factors of the species, as documented by other studies: very often MPs do not accumulate in the stomach and are expelled from the individual.