Different Lengths of Diet Supplementation with 10% Flaxseed Alter the Hormonal Profile and the Follicular Fluid Fatty Acid Content of Fattening Gilts.

The effect of 10% dietary flaxseed fed for 3 and 6 weeks on serum hormone levels of fattening gilts, the fatty acid (FA) follicular fluid (FF) composition of small and large antral follicles, and the steroidogenesis and IGF-I secretion by isolated small antral follicles and their response to regulatory hormones (LH, FSH, IGF-I) was studied using immunoassay and gas chromatography analyses. Both supplemental periods increased levels of P4 and IGF-I in blood serum. A shorter period inhibited steroidogenesis (P4, T, E2) and IGF-I secretion by small antral follicles, which was associated with decreased levels of monounsaturated FAs (MUFA) and preferred n-6 polyunsaturated FA (PUFA) metabolism. A longer period stimulated hormone secretion at elevated levels of saturated FAs (SFA) at the expense of MUFAs and PUFAs preferring the n-3 PUFA metabolism. Out of ovarian regulators, only LH and IGF-I were able to alter the secretion of steroids and IGF-I by small follicles of fattening pigs fed a basal diet. The effect of flaxseed on the secretion of follicular hormones after both supplemental periods was altered by all regulatory hormones in a dose-dependent manner. The level of SFAs and PUFAs in FF of large follicles increased with the length of flaxseed feeding, suggesting the suppression of ovulation.

Tetracalcium Phosphate Biocement Hardened with a Mixture of Phytic Acid-Phytase in the Healing Process of Osteochondral Defects in Sheep.

Hyaline articular cartilage has unique physiological, biological, and biomechanical properties with very limited self-healing ability, which makes the process of cartilage regeneration extremely difficult. Therefore, research is currently focused on finding new and potentially better treatment options. The main objective of this in vivo study was to evaluate a novel biocement CX consisting of tetracalcium phosphate-monetit biocement hardened with a phytic acid-phytase mixture for the regeneration of osteochondral defects in sheep. The results were compared with tetracalcium phosphate-monetit biocement with classic fast-setting cement systems and untreated defects. After 6 months, the animals were sacrificed, and the samples were evaluated using macroscopic and histologic methods as well as X-ray, CT, and MR-imaging techniques. In contrast to the formation of fibrous or fibrocartilaginous tissue on the untreated side, treatment with biocements resulted in the formation of tissue with a dominant hyaline cartilage structure, although fine fibres were present (p < 0.001). There were no signs of pathomorphological changes or inflammation. Continuous formation of subchondral bone and hyaline cartilage layers was present even though residual biocement was observed in the trabecular bone. We consider biocement CX to be highly biocompatible and suitable for the treatment of osteochondral defects.

Duration of the Flaxseed Supplementation Affects Antioxidant Defence Mechanisms and the Oxidative Stress of Fattening Pigs.

This study was conducted to investigate the effect of the duration of a flaxseed diet on fattening pigs' antioxidant defence mechanism in blood and tissues. Eighteen 20-week-old Landrace breed fattening pigs (BW 76.61 ± 2.30 kg) were divided into three groups of six animals. The control group was fed a basal diet. The FS3 group was fed the basal diet supplemented with 10% flaxseed for 3 weeks. The FS6 group received the same basal diet with flaxseed for 6 weeks. The total antioxidant capacity of the blood, measured as the total antioxidant status (TAS), total plasma antioxidant capacity (FRAP), reactive oxygen metabolites (dROMs) and total antioxidant capacity (PAT), was not affected by the flaxseed diet. The superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) activities were significantly decreased in the FS3 pigs in the heart (p < 0.05). However, in the FS3 group, the glutathione-S-transferase (GST) activity significantly increased compared to the control, but in the FS6 group, the activity was inhibited (p < 0.05). In the muscle, the CAT and GST activity was significantly decreased in the FS3 group (p < 0.05). The thiobarbituric acid reactive substance (TBARS) content was significantly reduced in the brain, muscle and heart in the FS3 group(p < 0.05). In FS6, the TBARS content significantly increased in the heart and brain (p < 0.05). Our results showed that the health effect of a flaxseed diet is significantly conditioned by the length of the flaxseed addition.

Beneficial Effect of Faecal Microbiota Transplantation on Mild, Moderate and Severe Dextran Sodium Sulphate-Induced Ulcerative Colitis in a Pseudo Germ-Free Animal Model.

Transplantation of faecal microbiota (FMT) is generally considered a safe therapeutic procedure with few adverse effects. The main factors that limit the spread of the use of FMT therapy for idiopathic inflammatory bowel disease (IBD) are the necessity of minimising the risk of infection and transfer of another disease. Obtaining the animal model of UC (ulcerative colitis) by exposure to DSS (dextran sodium sulphate) depends on many factors that significantly affect the result. Per os intake of DSS with water is individual for each animal and results in the development of a range of various forms of induced UC. For this reason, the aim of our study was to evaluate the modulation and regenerative effects of FMT on the clinical and histopathological responses and the changes in the bowel microenvironment in pseudo germ-free (PGF) mice of the BALB/c line subjected to chemical induction of mild, moderate and serious forms of UC. The goal was to obtain new data related to the safety and effectiveness of FMT that can contribute to its improved and optimised use. The animals with mild and moderate forms of UC subjected to FMT treatment exhibited lower severity of the disease and markedly lower damage to the colon, including reduced clinical and histological disease index and decreased inflammatory response of colon mucosa. However, FMT treatment failed to achieve the expected therapeutic effect in animals with the serious form of UC activity. The results of our study indicated a potential safety risk involving development of bacteraemia and also translocation of non-pathogenic representatives of bowel microbiota associated with FMT treatment of animals with a diagnosed serious form of UC.

Dietary Supplementation of Flaxseed (Linum usitatissimum L.) Alters Ovarian Functions of Xylene-Exposed Mice.

The aim of the performed study was to examine the ability of xylene, flaxseed, and their combinations to affect morphological and endocrine indexes of murine ovaries. The 72 indexes of secondary and tertiary follicular cells, oocytes, corpora lutea, and ovarian stroma have been quantified: diameter, markers of proliferation PCNA and apoptosis caspase 3, receptors to FSH, oxytocin, estrogen (alpha and beta), and progesterone. In addition, concentrations of the ovarian hormones progesterone, estradiol, and IGF-I in the blood, as well as their production by isolated ovaries cultured with and without gonadotropins (FSH + LH mixture), were determined using histological, immunohistochemical, and immunoassay analyses. The character of xylene and flaxseed effects on ovarian functions in mice depended on the stage of ovarian folliculogenesis. It was shown that flaxseed could mitigate and prevent the major (63%) effects of xylene on the ovary. In addition, the ability of gonadotropins to affect ovarian hormone release and prevent its response to xylene has been shown. The effects of these additives could be mediated by changes in the release and reception of hormones. These observations suggest that flaxseed and possibly gonadotropins could be natural protectors of a female reproductive system against the adverse effects of xylene.

Effect of Concentration of Flaxseed (Linum usitatissimum) and Duration of Administration on Fatty Acid Profile, and Oxidative Stability of Pork Meat.

Flaxseed is a common ingredient used for livestock feed. The aim of this work was to study the effect of a diet supplemented with flaxseed at 5% and 10% concentrations in the intervals of 3 and 6 weeks prior slaughter on fatty acid profile and oxidative stability of pork meat. Meat samples were collected after slaughter from each animal (five groups, n = 6). Samples of the musculus longissimus dorsi (MLD) and the musculus gluteobiceps (MGB) were selected. Chemical composition, fatty acid profile and oxidative stability during the storage of meat under chilling conditions (4 °C, 7 days) was analyzed. The addition of flaxseed significantly affected the composition of fatty acid profile and the shelf life of the produced meat. The fat content was changed in the experimental groups with 10% flaxseed supplementation (10.84% in MGB and 9.56% MLD) versus the control group. Despite the different concentrations of flaxseed, the best EPA/AA ratio was observed in the experimental groups fed with flaxseed supplementation for 3 weeks. The worst oxidative stability of meat samples (p < 0.05) was recorded in the experimental groups with the addition of flaxseed for 6 weeks, which was related to higher PUFA content in samples of the experimental groups and higher susceptibility of PUFAs to lipid oxidation. The oxidative stability of meat in the experimental group fed 5% flaxseed supplementation for 3 weeks was not affected.

Effects of supplemental flaxseed on the ovarian and uterine functions of adult cycling mice.

Dietary fatty acids (FA) have an effect on animal reproduction. The purpose of the study was to find out how supplemental flaxseed can modulate the FA metabolism and how FA can influence the release of ovarian hormones and functions of the uteri through the modulation of their specific receptors. Cycling mice were fed a basal diet (control) and basal diet fortified with 10% flaxseed for 6 weeks to examine its influence on the structure and function of the ovaries and uteri, and serum levels of FA. Unlike controls (30%), 100% of mice fed flaxseed exposed oestrus at the end of the supplemental period. Serum FA were analysed using gas chromatography. The ovaries and uteri underwent histological and immunohistochemical analyses, respectively. The ovarian fragments were cultured with or without follicle-stimulating hormone and culture media were analysed for progesterone (P4) and oestradiol-17ß (E2) using immunoassays. Dietary flaxseed increased the serum FA concentration, sizes of the ovaries and primary follicles, the release of P4 and E2, the thickness of endometrium and myometrium, and altered the expression of oestrogen and progesterone receptors in all uterine compartments. Dietary flaxseed can promote ovarian steroidogenesis and uterine activity in the mouse.

The potential adverse effect of 2.45 GHz microwave radiation on the testes of prenatally exposed peripubertal male rats.

In utero development of organs is easily influenced by many environmental factors. The aim of this study was to elucidate the effect of microwave radiation (MR) at a frequency of 2.45 GHz and a specific absorption rate of 1.73 W/kg on intrauterine development of testis. Pregnant albino rats were exposed to whole-body MR for 2 hours per day throughout the pregnancy. Male offspring (n=12, age 35 days) were not exposed to MR after birth. The study revealed that MR applied in utero induced apparent structural changes in the testes, such as irregular shape of seminiferous tubules, significant decrease in the diameter of seminiferous tubules (p<0.05) and in the height of the germinal epithelium (p<0.01), disorganisation of germ cells, desquamations of immature germ cells, formation of giant multinucleated cells, and significant (p<0.01) expansion of the interstitium. At the level of transmission electron microscopy, there were observed basement membrane irregularities in seminiferous tubules, vacuolation of the cytoplasm and adversely affected organelles in Sertoli cells, germ cells, Leydig cells, peritubular and endothelial cells. The tight junctions between adjacent Sertoli cells were often incomplete, and necrotizing germ cells were more numerous in experimental animals compared to controls. Enhanced necrotizations of germ cells proved by a Fluoro Jade C method, and declined germ cells proliferation confirmed by proliferating cell nuclear antigen analysis, were detected in MR exposed animals. Our results revealed that the prenatal exposure to MR had an adverse effect on the postnatal testicular development in rats.

In Vivo Study of Osteochondral Defect Regeneration Using Innovative Composite Calcium Phosphate Biocement in a Sheep Model.

This study aimed to clarify the therapeutic effect and regenerative potential of the novel, amino acids-enriched acellular biocement (CAL) based on calcium phosphate on osteochondral defects in sheep. Eighteen sheep were divided into three groups, the treated group (osteochondral defects filled with a CAL biomaterial), the treated group with a biocement without amino acids (C cement), and the untreated group (spontaneous healing). Cartilages of all three groups were compared with natural cartilage (negative control). After six months, sheep were evaluated by gross appearance, histological staining, immunohistochemical staining, histological scores, X-ray, micro-CT, and MRI. Treatment of osteochondral defects by CAL resulted in efficient articular cartilage regeneration, with a predominant structural and histological characteristic of hyaline cartilage, contrary to fibrocartilage, fibrous tissue or disordered mixed tissue on untreated defect (p < 0.001, modified O'Driscoll score). MRI results of treated defects showed well-integrated and regenerated cartilage with similar signal intensity, regularity of the articular surface, and cartilage thickness with respect to adjacent native cartilage. We have demonstrated that the use of new biocement represents an effective solution for the successful treatment of osteochondral defects in a sheep animal model, can induce an endogenous regeneration of cartilage in situ, and provides several benefits for the design of future therapies supporting osteochondral defect healing.

Dietary flaxseed's protective effects on body tissues of mice after oral exposure to xylene.

Xylene is a common pollutant in the environment that enters the body of animals and humans in various ways, but most often through the respiratory tract and adversely affects their overall health. However, xylene effects after oral exposure have not been sufficiently studied. This study aimed to investigate the effects of xylene exposure on the mouse organism and to identify possible beneficial effects of flaxseed on such exposure. Eighty mice were divided into four groups: control group C (basal diet + no xylene exposure), group X (oral exposure by 400 mg/kg/day xylene), group F (10% flaxseed supplementation of basal diet), and group XF (10% dietary flaxseed + oral exposure by xylene). Experimental trial took 14 days. Clinical examination, spectroscopic analysis of tissue aminotransferases, total lactate dehydrogenase (TLDH), and acetylcholinesterase (AchE) activities, electrophoretic analysis of LDH isoenzymes, western blot and immunohistochemical analysis of apoptosis as well as routine histology of the kidneys and jejunum, and transmission electron microscopy of the liver were performed. Marked restlessness in group X and high weight losses in mice of all groups were recorded during the experiment. Xylene promoted apoptosis (caspase-3 expression) without causing marked structural changes in the liver and jejunum, although renal cortex structure was affected adversely. In the brain, liver, and kidney of mice, xylene increased levels of liver transaminases, LDH, and decreased AchE activities, reflecting cell membrane damage. Flaxseed feeding improved animal behaviour, leakage of enzymes and prevented selected tissue toxic damage induced by xylene by protecting cell membrane integrity and fluidity and by suppressing apoptosis. These results point at the protective effect of flaxseed consumption on mice.

A Comprehensive Study Monitoring the Venom Composition and the Effects of the Venom of the Rare Ethiopian Endemic Snake Species Bitis parviocula.

The Ethiopian endemic snake of the species Bitis parviocula, recognized for its colorful patterns, might be more interesting as we look deeper into the venom activity. We assayed the effects of venoms from the most widespread venomous African Bitis arietens and closely related species Bitis parviocula using The Hen's Egg Test-Chorioallantoic membrane test (HET-CAM) and Chicken embryotoxicity screening test (CHEST), acetylcholinesterase (AChE) analysis, cytotoxicity assay performed on cell lines and protein analysis of selected venoms. Our results indicated that B. parviocula venom contains vasoactive compounds that have a direct effect on blood vessels. The AChE analysis showed significant ability inhibiting AChE activity in embryonic tissue. Cytotoxicity observed on A549 ATCC® CCL-185™ cells indicates the possible presence of cytotoxic agents in B. parviocula venom. We proved previously described differences in the composition of venom obtained from B. arietans and B. parviocula by using electrophoresis and total protein concentration. Based on similarities in vasoactive effects observed after administration of venoms onto a chicken chorioallantoic membrane, we suggest that venom from B. arietans and B. parviocula might share certain venom proteins responsible for haemotoxicity. The main active components of B. parviocula venom are unknown. Our results suggest that it might be worth performing proteomic analysis of B. parviocula venom as it might contain medically valuable compounds.

Potential influence of prenatal 2.45 GHz radiofrequency electromagnetic field exposure on Wistar albino rat testis.

An ever-increasing use of wireless devices over the last decades has forced scientists to clarify their impact on living systems. Since prenatal development is highly sensitive to numerous noxious agents, including radiation, we focused on the assessment of potential adverse effects of microwave radiation (MR) on testicular development. Pregnant Wistar albino rats (3 months old, weighing 282±8 g) were exposed to pulsed MR at a frequency of 2.45 GHz, mean power density of 2.8 mW/cm², and a specific absorption rate of 1.82 W/kg for 2 hours/day throughout pregnancy. Male offspring were no longer exposed to MR following birth. Samples of biological material were collected after reaching adulthood (75 days). In utero MR exposure caused degenerative changes in the testicular parenchyma of adult rats. The shape of the seminiferous tubules was irregular, germ cells were degenerated and often desquamated. The diameters of the seminiferous tubules and the height of the germinal epithelium were significantly decreased (both at ∗∗p<0.01), while the interstitial space was significantly increased (∗∗p<0.01) when compared to the controls. In the group of rats prenatally exposed to MR, the somatic and germ cells were rich in vacuoles and their organelles were often altered. Necrotizing cells were more frequent and empty spaces between Sertoli cells and germ cells were observed. The Leydig cells contained more lipid droplets. An increased Fluoro Jade - C and superoxide dismutase 2 positivity was detected in the rats exposed to MR. Our results confirmed adverse effects of MR on testicular development.

Innovative Animal Model of DSS-Induced Ulcerative Colitis in Pseudo Germ-Free Mice.

The aim of this study was to investigate the use of a standardized animal model subjected to antibiotic treatment, and the effects of this treatment on the course of dextran sodium sulphate (DSS)-induced colitis in mice. By decontamination with selective antibiotics and observation of pathogenesis of ulcerative colitis (UC) induced chemically by exposure of mice to various concentrations of DSS, we obtained an optimum animal PGF model of acute UC manifested by mucin depletion, epithelial degeneration and necrosis, leading to the disappearance of epithelial cells, infiltration of lamina propria and submucosa with neutrophils, cryptitis, and accompanied by decreased viability of intestinal microbiota, loss of body weight, dehydration, moderate rectal bleeding, and a decrease in the selected markers of cellular proliferation and apoptosis. The obtained PGF model did not exhibit changes that could contribute to inflammation by means of alteration of the metabolic status and the induced dysbiosis did not serve as a bearer of pathogenic microorganisms participating in development of ulcerative colitis. The inflammatory process was induced particularly by exposure to DSS and its toxic action on compactness and integrity of mucosal barrier in the large intestine. This offers new possibilities of the use of this animal model in studies with or without participation of pathogenic microbiota in IBD pathogenesis.

Effect of Thymol Addition and Withdrawal on Some Blood Parameters, Antioxidative Defence System and Fatty Acid Profile in Rabbit Muscle.

Thymol concentrations in rabbit plasma, intestinal wall (IW) and faeces were detected, and the effects of thymol application and withdrawal on biochemical, antioxidant parameters and fatty acids (FA) in blood (B) and muscle (M) were studied. Forty-eight rabbits were divided into two experimental groups (control, C and with thymol 250-mg/kg feed, T). Thymol was administered for 21 days (TA) and withdrawn for seven days (TW). Thymol in plasma correlated with that in the IW (Spearman's correlation coefficient (rs) = -1.000, p = 0.0167, TA) and was detected in faeces (TA and TW). In TA alkaline phosphatase (p = 0.0183), cholesterol (p = 0.0228), malondialdehyde (p = 0.003), glutathione peroxidase (p = 0.0177) in B and lactate dehydrogenase (M, p = 0.0411) decreased; monounsaturated FA (p = 0.0104) and α-linolenic acid (p = 0.0227) in M increased. In TW urea (p = 0.0079), docosapentaenoic acid (p = 0.0069) in M increased; linoleic acid (p = 0.0070), ∑ n-6 (p = 0.0007) in M and triglycerides decreased (B, p = 0.0317). In TA and TW, the total protein (p = 0.0025 and 0.0079), creatinine (B; p = 0.0357 and 0.0159) and oleic acid (M; p = 0.0104 and 0.0006) increased. Thymol was efficiently absorbed from the intestine and demonstrated its biological activity in blood and the muscles.

3D Reconstruction and Evaluation of Accessory Hepatic Veins in Right Hemilivers in Laboratory Animals by Metrotomography: Implications for Surgery.

BACKGROUND The current study investigated the detection of accessory hepatic veins and their vascular territories in the right hemiliver in rats, guinea pigs, and rabbits, which has become a prerequisite for newly developed clinical procedures. We compared the anatomical continuity of accessory hepatic veins with accessory hepatic veins existing in human livers. MATERIAL AND METHODS The analysis of accessory hepatic veins was performed using a corrosion cast method in combination with computer tomography (CT). RESULTS In normal livers, accessory hepatic veins were regularly found. The length of these veins was 0.88±0.29 (cm ±SD) in rats, 1.10±0.39 in guinea pigs, and 1.28±0.48 in rabbits. Accessory hepatic veins became a part of the draining vessel draining into segment VI and VII; represented by interpolating and following Chouinard's segmental concept. CONCLUSIONS The importance of detecting accessory hepatic veins lies in the identification of structures requiring special attention during surgery, in reduction of surgical complications, and in choosing the best approach to maintain the vitality of a drainage segment. The vascular reconstruction should be done during surgical interventions.

Supplemental flaxseed modulates ovarian functions of weanling gilts via the action of selected fatty acids.

The aim of this study was to examine the influence of dietary flaxseed on the endocrine and ovarian functions of weanling gilts challenged with E. coli and Coronavirus infections treated with dietary probiotic cheeses and to understand the possible mechanisms of its effects on ovarian function. Probiotics were used as a natural substitution for antibiotics and 10% dietary flaxseed is an effective prebiotic which supports the action of probiotics and has other beneficial effects on the organism. Probiotics with or without flaxseed were fed to weanling gilts starting 10 days before and lasting up until 14 days after weaning. The ovaries were measured and histologically analysed. The blood samples for the levels of steroid hormones and insulin-like growth factor I (IGF-I) were assessed using immunoassays and the levels of fatty acids were assessed using gas chromatography. All samples were collected on the day of weaning and 14 days after weaning. On the day of weaning, increased levels of linoleic acid and IGF-I was associated with higher body weight. The steroid hormones were not affected by the diet. The conversion of alpha-linolenic acid (ALA) to timodonic (EPA) and cervonic (DHA) acids were lower compared to controls, and together with high levels of myristic, palmitic and palmitoleic acids was associated with the higher proliferation and lower apoptosis in the primordial, primary and secondary follicles; although the inhibition of the cell cycle was observed in relation to the low level of eicosadienoic acid. The high levels of ALA, EPA and DHA and the low levels of myristic, palmitic and palmitoleic acids may have been the effect of flaxseed feeding 14 days post-weaning and may have had a reverse effect on the proliferation and apoptosis of ovarian follicles. These data suggest that flaxseed may suppress the follicle development in weanlings via the stimulation of apoptosis and the inhibition of proliferation via the modulation of the metabolism of selected fatty acids.

Orbital venous pattern in relation to extraorbital venous drainage and superficial lymphatic vessels in rats.

The purpose of this study was to demonstrate the normal and variant anatomy of extraorbital and intraorbital venous drainage together with retroorbital communication, and determine the lymphatic drainage from the superficial orbital region with a potential outlet of lymphatic vessel into the venous bloodstream. The study of the venous system was carried out on 32 Wistar rats by using corrosion casts methods and radiography, while the lymphatic system was studied in 12 Wistar rats following ink injection. Superficially, orbital veins are connected with extraorbital veins running through angular vein of the eye and the superficial temporal vein, and via the pterygoid plexus with the maxillary vein, which provide readily accessible communication routes in the spread of infection. The extent of intraorbital and periorbital venous drainage was ensured by the dorsal and ventral external ophthalmic vein through the infraorbital vein, which together formed the principal part of the ophthalmic plexus. Venous drainage of the eyeball was carried out mainly by the vortex veins, ciliary veins and internal ophthalmic vein. The highest variability, first presented by differences in structural arrangement and formation of anastomoses, was observed within the ventral external ophthalmic vein (22 cases) and the medial vortex vein (10 cases). Four vortex veins, one vein in each quadrant of the eye, were observed in rats. The vortex vein located on the ventral side of the eyeball was occasionally found as two veins (in four cases) in the present study. The lymphatic vessel from the lower eyelid entered into the mandibular lymph centre, and from the upper eyelid entered into the superficial cervical lymph centre, but both drained into the deep cranial cervical lymph node. The direct entry of lymph entering the veins without passing through lymph nodes was not observed.

Dietary supplementation of yucca (Yucca schidigera) affects ovine ovarian functions.

Yucca (Yucca schidigera) is a popular medicinal plant due to its many positive effects on animal and human physiology, including their reproductive systems. To examine the effect of supplemental yucca feeding on sheep reproduction, including ovarian functions and their hormonal regulators, ewes were fed (or not fed, control) yucca powder (1.5 g/head/day, 30 days). Macromorphometric indexes of the oviduct, ovary, and ovarian folliculogenesis were measured. Reproductive hormone levels in the blood were measured using a radioimmunoassay. Granulosa cells were aspirated from the ovary, and their proliferation and apoptosis were detected using immunocytochemistry. To assess secretory activity and its response to gonadotropin, ovarian fragments of treated and control ewes were cultured with and without follicle-stimulating hormone (FSH; 0, 0.1, 1, 10, or 100 IU/mL), and the release of reproductive hormones into the culture medium was evaluated. Finally, to examine the direct action of yucca on the ovary, ovarian fragments from control ewes were cultured with and without yucca extract (1, 10, or 100 µg/mL), and the release of reproductive hormones was measured. Yucca supplementation significantly decreased the size of small antral follicles (2 to <5 mm in diameter), increased accumulation of the apoptosis marker bax, and decreased serum progesterone (P4) and estradiol (E2) levels. It inhibited the release of P4 (but not other hormones), to prevent the stimulatory action of FSH on P4 output and promoted insulin-like growth factor I (IGF-I) release by fragments cultured with FSH. However, yucca supplementation did not affect the size of larger follicles and number of follicles, volume and weight of ovaries, length and weight of oviducts, caspase 3 accumulation, cell proliferation, testosterone (T) or IGF-I serum levels, or T or E2 release by cultured ovarian fragments and their response to FSH. Yucca addition to culture medium inhibited P4 and IGF-I, but not T or E2 release at the lowest (1 µg/mL) dose, and stimulated P4, but not T, E2, or IGF-I release at the highest (100 µg/mL) dose. These data suggest that yucca supplementation can reduce small antral ovarian follicle development possibly via the stimulation of apoptosis of their granulosa cells, suppression of ovarian P4 and E2 release, and alteration of ovarian IGF-I output and ovarian response to gonadotropin. Thus, yucca can directly affect P4 and IGF-I release by ovine ovarian cells.

Proteomic Analyses of Agkistrodon contortrix contortrix Venom Using 2D Electrophoresis and MS Techniques.

Snake venom is a complex mixture of proteins and peptides which in the Viperidae is mainly hemotoxic. The diversity of these components causes the venom to be an extremely interesting object of study. Discovered components can be used in search for new pharmaceuticals used primarily in the treatment of diseases of the cardiovascular system. In order to determine the protein composition of the southern copperhead venom, we have used high resolution two dimensional electrophoresis and MALDI ToF/ToF MS-based identification. We have identified 10 groups of proteins present in the venom, of which phospholipase A2 and metalloprotease and serine proteases constitute the largest groups. For the first time presence of 5'-nucleotidase in venom was found in this group of snakes. Three peptides present in the venom were also identified. Two of them as bradykinin-potentiating agents and one as an inhibitor.

Proteome and Peptidome of Vipera berus berus Venom.

Snake venom is a rich source of peptides and proteins with a wide range of actions. Many of the venom components are currently being tested for their usefulness in the treatment of many diseases ranging from neurological and cardiovascular to cancer. It is also important to constantly search for new proteins and peptides with properties not yet described. The venom of Vipera berus berus has hemolytic, proteolytic and cytotoxic properties, but its exact composition and the factors responsible for these properties are not known. Therefore, an attempt was made to identify proteins and peptides derived from this species venom by using high resolution two-dimensional electrophoresis and MALDI ToF/ToF mass spectrometry. A total of 11 protein classes have been identified mainly proteases but also l-amino acid oxidases, C-type lectin like proteins, cysteine-rich venom proteins and phospholipases A2 and 4 peptides of molecular weight less than 1500 Da. Most of the identified proteins are responsible for the highly hemotoxic properties of the venom. Presence of venom phospholipases A2 and l-amino acid oxidases cause moderate neuro-, myo- and cytotoxicity. All successfully identified peptides belong to the bradykinin-potentiating peptides family. The mass spectrometry data are available via ProteomeXchange with identifier PXD004958.