NEUROPROTECTIVE ANTIOXIDANT EFFECT OF PORTULACA QUADRIFIDA LINN. LEAVES EXTRACT ON IMMOBILIZATION STRESS-INDUCED CHANGES IN RAT'S BRAIN

Objective: The current study was established to illustrate the effect of this plant's active constituents on stress-induced diseases that include oxidation damage to cellular components, especially the brain.Methods: Immobilization stress method is induced and initiated by putting separated animals in specially prepared mesh cages on a wooden plank. The animals were exposed to 6 h of stress. Wistar male rats were selected weighing (180–200 g). A total of 54 rats were selected and separated into nine groups and only six experimental rats were kept in each group.Results: The post-stress oral treatment of extract (100 mg/kg body weight) was more efficient in restricting stress-induced decline of superoxide dismutase (p<0.05), glutathione (GSH)-S-transferase (p<0.02), catalase (p<0.05), and GSH (p<0.05) and an enhanced level of thiobarbituric acid reactive substance (p<0.01) compared to stress alone or pre-stress extract treatments.Conclusion: The extract showed a significant resistance toward the oxidative metabolism triggered by restraint stress, though the post-extract treatment (curative) was observed to be more effective in restoring the altered oxidative metabolism compared with pre-extract treatment (prophylactic)

Vitamin D and Obesity

Obesity is a very common issue worldwide, and it is one of the risk factors for mortality. Several studies were done to identify the causes of this issue and to investigate factors that can affect this condition. Vitamin D is claimed to have an impact not only for maintaining bone health but also for having an association between its deficiency and obesity as some studies found that the concentrations of this vitamin are low in obese individuals. The suggested mechanisms and a discussion of the latest findings as well as the possibility of integrating supplementation in the treatment of obesity are covered in this book chapter. It was concluded that vitamin D deficiency is prevalent in many parts of the world and the supplements are an affordable option, but further studies are required to address different confounding factors that will result in clear data interpretation and will contribute to the future planning of health policies and guidelines used by healthcare professionals

VITAMIN D IMPACT ON HUMAN HEALTH AND ITS RELATION WITH SEVERAL DISEASES

Vitamins are organic nutrients and substances that are needed for many biochemical functions. They are one of the six essential elements required for human life. Vitamin D is also known among the public as the Sunshine vitamin, as it is produced internally within the human body when exposed to sunlight and it is well known that the big majority of United Arab Emirates (UAE) population suffer from its deficiency.They are usually not found inside the body and therefore must be eaten from food or taken by supplements. Vitamin D that is obtained from diet or body synthesis when it is exposed to sufficient sunlight is biologically inactive without the enzymatic activation that converts it into its active form in the liver and kidney.Deficiency of vitamin D could lead to many troubling diseases such as depression including seasonal affective disorder, Parkinson's disease, Alzheimer's disease, Arthritis (gout, osteoarthritis, tendonitis), Osteoporosis, gum and teeth diseases, obesity, diabetes, heart diseases, metabolic syndromes, immunity related diseases (sclerosis, erythematosis), and cancer, which will be discussed in this review. The main objective of the current review is to evaluate the associations of vitamin D with the diverse health outcomes according to the latest studies done so far

Assessment of knowledge and healthcare providers’ role in promoting lactation in United Arab Emirates

Background: This study was performed to assess mothers’ knowledge and awareness level regarding maternal medication intake during lactation and health care providers’ role in promoting lactation.Methods: This study is a cross-sectional, questionnaire based survey study and a convenient sample of (820) breastfeeding mothers in Ajman and Sharjah, United Arab Emirates (U.A.E.) participated in the study. The results were analyzed using the Statistical Package for the Social Sciences (SPSS) version 20. Descriptive statistics were used to summarize the data concerning the demographic characteristics and breastfeeding knowledge. Categorical variables (such as nationality and educational level) were described by using frequency, percentages, bar chart and pie chart.Results: The current study revealed a (72%) of maternal knowledge compared to (60.4%) four years ago and the most identified encouraging factor for the initiation of lactation stated by the respondents was their own decision followed by family support (58.4%, 39.8% respectively). The health care providers’ role was positively improved (58.4%) as observed in the current study compared to (52.1%) four years ago in UAE.Conclusions: An improvement was observed in terms of knowledge among breastfeeding mothers in the UAE compared to previous years as well as healthcare providers’ role which was also positively improved. Pharmacists were observed to have a low impact on the initiation of lactation, but on the other hand it was found that they had a significant role in terms of assessing maternal medication safety during lactation

Optimization of CASA-Mot Analysis of Donkey Sperm : optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity-VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis

Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620-630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects

The Effects of Red Light on Mammalian Sperm Rely upon the Color of the Straw and the Medium Used

Several studies have shown that the exposure of semen to red light improves sperm quality and fertilizing ability, which could improve the efficiency of assisted reproductive techniques with irradiated semen. However, despite being considered as possible sources of variation, the effects of the color of the container (straws) or the medium have not yet been evaluated. In this study, 13 ejaculates from different stallions were split into equal fractions, diluted either with Kenney or Equiplus extender, and subsequently packed into straws of five different colors. After storage at 4 °C for 24 h, the sperm were irradiated and different variables, including sperm motility, plasma membrane integrity, and mitochondrial membrane potential, were evaluated. Our results confirm that irradiation increases some motion characteristics and mitochondrial membrane potential without affecting sperm viability and demonstrate that the effects depend on the color of the straw and the extender used. Previous research has determined that irradiation of mammalian sperm with red light increases motility, mitochondrial activity, and fertilization capacity. In spite of this, no study has considered the potential influence of the color of the straw and the extender used. Therefore, this study tests the hypothesis that the response of mammalian sperm to red light is influenced by the color of the straw and the turbidity/composition of the extender. Using the horse as a model, 13 ejaculates from 13 stallions were split into two equal fractions, diluted with Kenney or Equiplus extender, and stored at 4 °C for 24 h. Thereafter, each diluted fraction was split into five equal aliquots and subsequently packed into 0.5-mL straws of red, blue, yellow, white, or transparent color. Straws were either nonirradiated (control) or irradiated with a light-dark-light pattern of 3-3-3 (i.e., light: 3 min, dark: 3 min; light: 3 min) prior to evaluating sperm motility, acrosome and plasma membrane integrity, mitochondrial membrane potential, and intracellular ROS and calcium levels. Our results showed that irradiation increased some motion variables, mitochondrial membrane potential, and intracellular ROS without affecting the integrities of the plasma membrane and acrosome. Remarkably, the extent of those changes varied with the color of the straw and the extender used; the effects of irradiation were more apparent when sperm were diluted with Equiplus extender and packed into red-colored straws or when samples were diluted with Kenney extender and packed into transparent straws. As the increase in sperm motility and intracellular ROS levels was parallel to that of mitochondrial activity, we suggest that the impact of red light on sperm function relies upon the specific rates of energy provided to the mitochondria, which, in turn, vary with the color of the straw and the turbidity/composition of the extender

Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis

Altres ajuts: Generalitat de Catalunya 2017-SGR-1229Jenny shows a large endometrial reaction after semen influx to the uterus with a large amount of polymorphonuclear neutrophils (PMN) migrating into the uterine lumen. PMN act as a sperm selection mechanism through phagocytosis and NETosis (DNA extrudes and, together with proteins, trap spermatozoa). While a reduced percentage of spermatozoa are phagocytosed by PMN, most are found to be attached to neutrophil extracellular traps (NETs). This selection process together with sperm metabolism produces a large amount of reactive oxygen species (ROS) that influence the reproductive success. The present study aimed to determine the extracellular ROS production in both sperm and PMN. With this purpose, (1) donkey sperm were exposed to reductive and oxidative stresses, through adding different concentrations of reduced glutathione (GSH) and hydrogen peroxide (HO), respectively; and (2) PMN were subjected to NETosis in the presence of the whole semen, sperm, seminal plasma (SP) or other activators such as formyl-methionyl-leucyl-phenylalanine (FMLP). Extracellular ROS production (measured as HO levels) was determined with the Amplex ® Red Hydrogen Peroxide/Peroxidase Assay Kit. Donkey sperm showed more resilience to oxidative stress than to the reductive one, and GSH treatments led to greater HO extracellular production. Moreover, not only did SP appear to be the main inducer of NETosis in PMN, but it was also able to maintain the extracellular HO levels produced by sperm and NETosis

Computerised analysis of semen in equids

L'enfocament que es proposa en aquesta tesi es basa en el fet que hi ha un buit d'informació sobre l'optimització de l'examen de la solidesa reproductiva juntament amb els protocols d'anàlisi de semen en els equids, tal com es va veure al capítol anterior. A l'ase, no es van definir protocols específics per a l'anàlisi de semen, només per a mostres de sementals on hi ha un acord general sobre el protocol a utilitzar. Quan es comprova el tracte reproductor, s'utilitza una ecografia en mode B com a mètode de diagnòstic fàcil i no invasiu. D'altra banda, es pot utilitzar una ecografia Doppler polsada per estudiar la perfusió testicular de sang. Els índexs de Doppler pulsat de l'artèria testicular i les mides ASG van mostrar una correlació positiva amb la qualitat del semen en sementals i diferents espècies. Malauradament, no es van fer estudis d'aquest tipus en ruc, cosa que constitueix el primer objectiu de la tesi. No obstant això, quan es tracta d'anàlisi de semen, la tecnologia CASA representa el millor mètode objectiu que s'utilitza avui en dia. Permet obtenir dades quantitatives fiables, fins i tot si és necessari definir protocols que assegurin la consistència i l'aplicació universal dels resultats. Tanmateix, aquesta estandardització mai s'ha fet seguint un punt de vista integrador, que compon el segon objectiu general. Hi ha tres aspectes principals a tenir en compte a l'hora d'optimitzar les anàlisis automatitzades de semen mitjançant la tecnologia CASA-Mot, a saber, el tipus, la profunditat de la cambra de comptatge, la dilució i la velocitat de fotogrames d'adquisició d'imatges. Els avenços tecnològics van permetre ara l'adquisició d'alta freqüència de captura i, al mateix temps, el procés d'aquestes imatges en segons. A més, es va definir l'estructura de les subpoblacions d'espermatozoides en marcs més alts i la distribució SP entre les races dels sementals. El tercer objectiu era col·laborar en l'ús i la millora de la nova tècnica Trumorph®, per a l'anàlisi de morfologia de l'esperma que permet l'anàlisi d'espermatozoides immobilitzats vius.El enfoque defendido en esta tesis se basa en el hecho de que existe un vacío de información con respecto a la optimización del examen de solidez reproductiva junto con los protocolos de análisis de semen en équidos, como se vio en el capítulo anterior. En burro, no se definieron protocolos específicos para el análisis de semen, solo para muestras de sementales donde existe un acuerdo general sobre el protocolo a utilizar. Cuando se comprueba el tracto reproductivo, la ecografía en modo B se utiliza como un método de diagnóstico fácil y no invasivo. Por otro lado, la ecografía Doppler pulsado se puede utilizar para estudiar la perfusión sanguínea testicular. Los índices de Doppler pulsado de la arteria testicular y los tamaños de ASG mostraron una correlación positiva con la calidad del semen en sementales y diferentes especies. Lamentablemente, estos estudios no se realizaron en burro, lo que constituye el primer objetivo de la tesis. Sin embargo, cuando se trata de análisis de semen, la tecnología CASA representa el mejor método objetivo utilizado en la actualidad. Permite obtener datos cuantitativos confiables, incluso si es necesario para definir protocolos asegurando la consistencia y aplicación universal de los resultados. Sin embargo, esta estandarización nunca se ha hecho siguiendo un punto de vista integrador, que compone el segundo objetivo general. Hay tres aspectos principales a considerar al optimizar los análisis de semen automatizados mediante la tecnología CASA-Mot, a saber, el tipo, la profundidad de la cámara de recuento, la dilución y la velocidad de cuadros de la adquisición de imágenes. Los avances en tecnología permitieron ahora la adquisición de alta frecuencia de captura y al mismo tiempo el proceso de estas imágenes en segundos. Además, se definió la estructura de las subpoblaciones de espermatozoides en cuadros más altos y la distribución de SP entre las razas de sementales. El tercer objetivo fue colaborar en el uso y la mejora de la nueva técnica Trumorph®, para el análisis de la morfología espermática que permite el análisis de espermatozoides inmovilizados vivos.The approach advocated in this thesis is based on the fact that there is a gap of information regarding optimization of breeding soundness examination along with semen analysis protocols in equids as seen in the previous chapter. In donkey, no specific protocols for semen analysis were defined, only for stallion samples where there is a general agreement about the protocol to use. When the reproductive tract is checked, B-mode ultrasonography is used as a non-invasive and easy diagnostic method. On the other hand, pulsed Doppler ultrasonography can be used to study testicular blood perfusion. Pulsed-Doppler indices of testicular artery and ASG sizes showed a positive correlation with semen quality in stallion and different species. Unfortunately, no such studies were done in donkey, which makes up the first objective of the thesis. However, when it's about semen analysis, CASA technology represents the best objective method used today. It permits to obtain reliable quantitative data, even if it is needed to define protocols assuring the consistency and universal application of the results. However, this standardization has never been done following an integrative point of view, which composes the second general objective. There are three main aspects to consider when optimizing automated semen analyses by CASA-Mot technology, namely the type, depth of the counting chamber, the dilution, and the frame rate of image acquisition. The advances in technology allowed now the acquisition of high capture frequency and at the same time the process of these images in seconds. Also, was defined sperm subpopulations structure at higher frames and SP distribution between stallions' breeds. The third objective was to collaborate in the use and the improvement of the new Trumorph® technique, for sperm morphology analysis which permits the analysis of alive immobilized sperm

New Sperm Morphology Analysis in Equids : Trumorph ® Vs Eosin-Nigrosin Stain

Equine Reproduction Service. Department of Animal Medicine and Surgery. Veterinary Faculty. Autonomous University of Barcelona. National Agency for Research and Development (ANID), Ministry of Education, Chile (2017/72180128). Secretary of Higher Education, Science, Technology and Innovation (SENESCYT)(CZ02-000507-2019)The evaluation of the male fertility potential is based on the analysis of the basic spermatic characteristics of concentration, motility and morphology. Thus, the study of sperm morphology is a fundamental element in the seminal analysis, but its real meaning has been biased by the techniques used for its evaluation. These techniques involve dehydration phases and subsequent staining, which involves the production of artifacts. The aim of the study is to compare two methods for equid semen morphology evaluation, Trumorph ® using living sperm vs. eosin-nigrosine stain. A total of 49 ejaculates from stallions and donkeys were used. After semen collection and dilution, an aliquot was placed on the slide and introduced in the Trumorph ® device. Then observation was made with a 40x objective and negative phase-contrast microscope. Another aliquot was stained using eosin-nigrosine stain and viewed using 100× magnification. Well-formed sperm were observed, and different abnormalities were identified using Trumorph ®. The use of eosin-nigrosin staining method and Trumorph ® led to the same results and both techniques can be used for stallion and donkey sperm morphological analysis. However, considering the fact that Trumorph ® uses living sperm helps prevent sperm cell alteration during sample preparation. Therefore, Trumorph ® can be a good alternative to the conventional staining method, which provides a quick test on live sperm