Optimization of spray-dried hyaluronic acid microspheres to formulate drug-loaded bone substitute materials

Wepresent here our first results concerning the evaluation of hyaluronic acid (HA) as a candidate to formulate an organic–mineral cement with sustained release properties. Incorporating drug-loaded microspheres in mineral bone cements is an alternative strategy to improve their ability as drug delivery materials. To synthesize microspheres according to a reproducible process and control at the same time their morphology and their encapsulation efficiency is one of the main challenges of the conception of such drug-loaded bone substitute. In this context, we investigated the potentialities of two HA, differing by theirmolecular weight, to form microspheres by a spray-drying technique. Erythrosin B (EB) was encapsulated as a model drug and spray-drying process conditionswere optimized. To performthis, the rheological behavior and viscosity of HA solutions have been related to their spray-drying ability, and then to the resulting microparticles morphological properties and size distribution. Reproducible microspheres, answering to the requirements in terms of size and encapsulation efficiency, have been obtained from both HA. However the HA exhibiting the lowest molecular weight, HA600, led to smaller microparticles, with a higher polydispersity index. Their swelling ability, related to their stability upon rehydration, also appeared reduced. In this context, HA850, with the highest molecular weight, was selected and the possibility to modulate drug release by HA850 microspheres incorporation into a mineral cement was explored. EB release kinetics from HA microspheres, HA microspheres loaded cement and reference cement were followed at 37 °C, in Tris buffer at pH 7.4, using European Pharmacopoeia flow-through cells. Results showed that HA microspheres incorporation into a mineral cement permitted to modify the cement drug release profile and led to a sustained release

Daily rhythms of behavioral and hormonal patterns in male dromedary camels housed in boxes

Background. Daily rhythmicity has been observed for a number of hormonal and behavioral variables in mammals. It can be entrained by several external factors, such as light-dark cycle and scheduled feeding. In dromedary camels, daily rhythmicity has been documented only for melatonin secretion and body temperature. In this study, the daily rhythmicity of behavioral repertoire, cortisol and testosterone levels was investigated in captive male camels. Methods. Six clinically healthy male dromedary camels (Camelus dromedarius) were used. The animals were housed in single boxes for 24 h daily and fed twice a day. Over a period of 48 h, behavioral observations were made and blood samples taken every two hours. The data were analyzed using diurnality index, conisor analysis and PROC mixed procedure. Results. The diurnality index for rumination and lying down was close to 0 (respectively, 0.09 and 0.19), while the indices for stereotypy, standing, feeding and walking were close to 1 (respectively, 0.74, 0.84, 0.92 and 0.85). Cosinor analysis revealed daily rhythmicity for all behaviors and for cortisol levels (acrophase at 12:57) but not for testosterone. Rumination and lying down (inactive behaviors) reached a peak during the scotophase, whereas feeding, walking and stereotypy (active behaviors) reached a peak during the photophase around midday. Cortisol level and expression of stereotypies peaked before and after food distribution and were negatively correlated (r =-0.287, P = 0.005). Testosterone levels and expression of sexual behaviors were stimulated by the visual and olfactory contacts with the females and were positively correlated (r =0.164, P =0.040). Testosterone was also negatively correlated with cortisol (r =-0.297; P =0.003). Discussion. These preliminary results provided new knowledge about the daily rhythm of behaviors in camels housed in boxes, suggesting that camels exhibit diurnal behavior pattern in the maintenance conditions outlined in the study. Daily rhythmicity seemed to be entrained not only by the light-dark cycle but also by scheduled feeding. The rise in stereotypy after food distribution could be due to the persistence of feeding motivation and frustration after the ingestion of food. Therefore, feeding practices should be improved to satisfy the foraging and feeding motivation of these camels. Behavioral and hormonal daily patterns in camels should be taken in consideration to adapt the management system, giving the animals more freedom during the light period and a diet richer in fiber, so as to improve reproductive performance, health and welfare

Quantitative Analysis of Semicrystalline Blends SAXS Data: Theoretical Modeling versus Linear Correlation Function

This paper compares the results yielded by two methods of small-angle X-ray scattering data analysis for semicrystalline polymer blends. The first method is based on the use of a theoretical modeling for isotropic samples and a subsequent curve fitting. The second one is a more familiar method, based on the calculation of the linear one-dimensional correlation function. The experimental material considered for this purpose deals with a series of semi-crystalline blends of poly(vinylidene fluoride) and poly(methyl methacrylate), with a PVDF content covering the range 50 wt%–100 wt%. The results obtained by both calculation methods are systematically confronted to the crystallinity degrees deduced from wide angle X-ray scattering patterns

Identification of ß-nerve growth factor in dromedary camel seminal plasma and its role in induction of ovulation in females

The main objective of this study was to demonstrate the effect of seminal plasma ovulation inducing factor (OIF) on ovulation in female camels. Seminal plasma was fractionated using gel filtration chromatography, and two protein peaks were obtained and analysed by western blotting. The effect of the bioactive protein fraction (OIF/ß-NGF) was tested by intramuscular injection (1 ml) in synchronized females with the following treatments: PBS (negative control group, n=3); 20 /jg Buserelin (GnRH analogue, positive control group; n=3); and purified OIF with doses of 1 and 2 mg (n = 5, each treatment). Blood samples were collected every two days from day 0 until 14 days post-treatment, and the progesterone concentration was assessed. The obtained results showed that the OIF is highly present in the seminal plasma of dromedary camels as a protein with a molecular mass of approximately 14 kDa. It was detected as a betaNerve Growth Factor, named Cam-ß-NGF. The effect of this molecule on ovulation was clearly demonstrated by the significant increase of the plasma progesterone concentration in the treated female groups (1 and 2 mg of Cam-ß-NGF) as observed in Buserelin group. In summary, intramuscular injection of ß-NGF isolated from dromedary camel seminal plasma induces ovulation in females with similar rate to Buserelin treatment

Development of an injectable composite for bone regeneration

With the development of minimally invasive surgical techniques, there is a growing interest in the research and development of injectable biomaterials especially for orthopedic applications. In a view to enhance the overall surgery benefits for the patient, the BIOSINJECT project aims at preparing a new generation of mineral-organic composites for bone regeneration exhibiting bioactivity, therapeutic activity and easiness of use to broaden the application domains of the actual bone mineral cements and propose an alternative strategy with regard to their poor resorbability, injectability difficulties and risk of infection. First, a physical-chemical study demonstrated the feasibility of self-setting injectable composites associating calcium carbonate-calcium phosphate cement and polysaccharides (tailor-made or commercial polymer) in the presence or not of an antibacterial agent within the composite formulation. Then, bone cell response and antimicrobial activity of the composite have been evaluated in vitro. Finally, in order to evaluate resorption rate and bone tissue response an animal study has been performed and the histological analysis is still in progress. These multidisciplinary and complementary studies led to promising results in a view of the industrial development of such composite for dental and orthopaedic applications

Mating behaviour and semen parameters in dromedary camel bulls (Camelus dromedarius): A comparison between two types of artificial vagina

Semen collection in the dromedary camel (Camelus dromedarius) is performed using different types of artificial vagina (AV) (long, short, with cervix imitation, with plastic or silicone inner liner) either held by an operator or fitted inside a camel dummy. However, systematic studies about the acceptance of such modifications by bulls, and their effects on sperm parameters, have yet to be investigated. This study was conducted to test the hypothesis that a silicone inner liner inserted into a standard cow AV could cause discomfort to camel bulls during mating, but at the same time could improve the quality of the ejaculates collected. Five bulls underwent semen collection sessions, with (n=3) and without (n=3) the silicone liner fitted in the AV, for a total of 30 semen collection sessions. A standard methodology and data-recording system were used for evaluating mating behavior, in addition to evaluation of semen parameters. There was no difference between an AV with or without silicone inner liner used during our standardized semen collection procedure. The use of such an AV modification hence still remains debatable, until further studies are able to identify any potential benefits of using this or other AV types or modifications. Such studies, however, should be performed by applying a standardized semen collection methodology and a proper evaluation of mating behaviour and semen parameters

Estimation of the economic impact of a bluetongue serotype 4 outbreak in Tunisia

IntroductionSince 1999, Tunisia has experienced multiple occurrences of Bluetongue (BT) outbreaks, leading to numerous reported cases of infection and mortality in flocks. The re-emergence of the disease in 2020 caused substantial economic losses in cattle, attributed to the incursion of serotype BTV-4.MethodsTo evaluate the economic impact of the recent BT episode, we conducted a retrospective study on outbreaks that occurred in Tunisia between August and November 2020, focusing on the impact at the owner’s level and its effects on both small ruminants and cattle. A total of 234 ruminant farms (sheep, cattle, and mixed) were randomly selected across Tunisian governorates and included in the study to estimate both the direct and indirect costs of these outbreaks.ResultsTotal costs were calculated as the sum of losses and expenditures resulting from the BT outbreaks. At the animal level, total losses were estimated to range between 116.280 and 207.086 TND for one infected ewe (€33.721 and 60.055). For one lactating cow, costs varied between 2,590.724 and 3,171.107 TND (€751.310 and 919.621). In cattle, exposure to BTV led to a daily unit milk yield decrease of 12.50 to 14.66 L over an average period of 5 months. Diseased sheep experienced weight loss ranging between 4 and 10 kg during the BT outbreaks. The total mean cost of the 2020 BT outbreak in Tunisian investigated farms was estimated at 1,935 million TND (million €561.15) (range: 1,489 and 2,474 million TND; 431.81 and million €717.46). The most influential costs of the total BT outbreaks were the decrease in milk yield, mortality, and veterinary treatment.DiscussionThis study gives valuable insights on the economic impact of the incursion of a new serotype of BT in a naive population in Tunisia. Considering the substantial costs incurred, it is imperative that this disease receives increased attention from stakeholders, including animal owners, veterinary services, practitioners, and decision-makers

Implementation of the One Health approach to fight arbovirus infections in the Mediterranean and Black Sea Region: Assessing integrated surveillance in Serbia, Tunisia and Georgia

Background In the Mediterranean and Black Sea Region, arbovirus infections are emerging infectious diseases. Their surveillance can benefit from one health inter-sectoral collaboration; however, no standardized methodology exists to study One Health surveillance. Methods We designed a situation analysis study to document how integration of laboratory/clinical human, animal and entomological surveillance of arboviruses was being implemented in the Region. We applied a framework designed to assess three levels of integration: policy/institutional, data collection/data analysis and dissemination. We tested the use of Business Process Modelling Notation (BPMN) to graphically present evidence of inter-sectoral integration. Results Serbia, Tunisia and Georgia participated in the study. West Nile Virus surveillance was analysed in Serbia and Tunisia, Crimea-Congo Haemorrhagic Fever surveillance in Georgia. Our framework enabled a standardized analysis of One Health surveillance integration, and BPMN was easily understandable and conducive to detailed discussions among different actors/institutions. In all countries, we observed integration across sectors and levels except in data collection and data analysis. Data collection was interoperable only in Georgia without integrated analysis. In all countries, surveillance was mainly oriented towards outbreak response, triggered by an index human case. Discussion The three surveillance systems we observed prove that integrated surveillance can be operationalized with a diverse spectrum of options. However, in all countries, the integrated use of data for early warning and inter-sectoral priority setting is pioneeristic. We also noted that early warning before human case occurrence is recurrently not operationally prioritized

Bacteria-inducing legume nodules involved in the improvement of plant growth, health and nutrition

Bacteria-inducing legume nodules are known as rhizobia and belong to the class Alphaproteobacteria and Betaproteobacteria. They promote the growth and nutrition of their respective legume hosts through atmospheric nitrogen fixation which takes place in the nodules induced in their roots or stems. In addition, rhizobia have other plant growth-promoting mechanisms, mainly solubilization of phosphate and production of indoleacetic acid, ACC deaminase and siderophores. Some of these mechanisms have been reported for strains of rhizobia which are also able to promote the growth of several nonlegumes, such as cereals, oilseeds and vegetables. Less studied are the mechanisms that have the rhizobia to promote the plant health; however, these bacteria are able to exert biocontrol of some phytopathogens and to induce the plant resistance. In this chapter, we revised the available data about the ability of the legume nodule-inducing bacteria for improving the plant growth, health and nutrition of both legumes and nonlegumes. These data showed that rhizobia meet all the requirements of sustainable agriculture to be used as bio-inoculants allowing the total or partial replacement of chemicals used for fertilization or protection of crops