Ultrastructural mechanical and material characterization of fossilized bone

Bone plays a key role in the paleontological and archeological records and can provide insight into the biology, ecology and the environment of ancient vertebrates. Examination of bone at the tissue level reveals a definitive relationship between nanomechanical properties and the local organic content, mineral content, and microstructural organization. However, it is unclear as to how these properties change following fossilization, or diagenesis, where the organic phase is rapidly removed and the remaining mineral phase is reinforced by the deposition of apatites, calcites, and other minerals. While the process of diagenesis is poorly understood, its outcome clearly results in the potential for dramatic alteration of the mechanical response of biological tissues. In this study, fossilized specimens of mammalian long bones, collected from Colorado and Wyoming, were studied for mechanical variations. Nanoindentation performed in both longitudinal and transverse directions revealed preservation of bone's natural anisotropy as transverse modulus values were consistently smaller than longitudinal values. Additionally, modulus values of fossilized bone from 35.0 to 89.1 GPa increased linearly with logarithm of the sample's age. Future studies will aim to clarify what mechanical and material elements of bone are retained during diagenesis as bone becomes part of the geologic milieu. © 2007 Materials Research Society.

A comparative study of various extenders of milkfish, Chanos chanos (Forsskal) sperm preservation

Four chemical extenders in 7 different concentrations (potassium chloride, sodium chloride, glucose, sodium citrate, Ringer s solution, cow serum and milkfish (Chanos chanos) serum) were compared in the preservation of milkfish sperm. Results showed milkfish serum to be the most suitable of the various extenders tested. This may be attributed to suitable osmotic potential and/or presence of proteins which may have directly or indirectly influenced sperm viability. The effects of milkfish serum on the motility and fertilizing capacity of sperm at different durations of storage however need to be investigated.

THE POSSIBLE INVOLVEMENT OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR IN LUTEOLYSIS OF RHESUS-MONKEY

Changes of plasminogen activators (PA) during different stages of development of the corpus luteum, and their possible physiological role in luteolysis were studied in rhesus monkeys. It was demonstrated for the first time that monkey corpus luteal cells not only produce PA, but that the function of the corpus luteum is also closely related to the activity of this enzyme system. Generally, the life span for a corpus luteum in monkey is approximately 14-16 days, its demise beginning thereafter. In the present study, we found that urokinase in the corpus luteum is higher on day 5 and day 10 after human chorionic gonadotrophin injection, while the tissue type (t) PA is mainly produced on day 13 when luteolysis may take place. Progesterone production remained high on day 5 and day 10 and decreased dramatically from day 13, indicating the important role of tPA but not urokinase (u) PA in suppressing luteal function. When purified tPA (but not uPA) monoclonal antibody was added to luteal cell culture to neutralize endogenously produced tPA activity, progesterone production in the cells was increased significantly. Interestingly, prolactin alone was capable of increasing PA production by luteal cells; prolactin together with luteinizing hormone, however, had a synergistic luteotrophic effect.

Effect of butachlor on spermatogenesis, testosterone amount and testes tissue Rutilus frisii kutum Kamenskii 1901

Morphological assessment of sexually mature Rutilus frisii kutum Kamenskii 1901 caught from the rivers (Shirud, Khoshkrud, Sepidrud and Chelavand Rivers) flowing in the southwest Caspian Sea region was conducted and sperm volume, total sperm count and sperm concentration of abnormal sperms were determined after exposing the spawners to 60% herbicide butachlor (machete). Spawners under study were maintained in tanks (1000 l) at the Shahid Ansari Teleost Fish Hatchery and exposed to two different concentrations (25% and 75% of its LC50 value) of butachlor. Results obtained indicate that exposure to high butachlor toxicity (75% of its LC50 value) decreased sperm volume to 0.61 ± 0.42 cc in 2-3 year old fishes and to 0.55 ± 0.42 cc in fishes above 3 years of age, while that in fish exposed to low butachlor toxicity (25% of its LC50 value) decreased to 1.55 ± 0.42 cc in 2-3 year old fishes and to 1.28 ± 0.42 cc in fishes above 3 years of age. The sperm volume under normal conditions in R. frisii kutum is 4.6 ± 0.42 cc in 2-3 year olds and 4.58 ± 0.42 cc in fishes above 3 years of age. The total sperm count in R. frisii kutum is 39.74 ± 2.5 billion spermatozoa/cc in 2-3 year olds and 42.99 ± 2.5 billion spermatozoa/cc in fishes above 3 years of age. When exposed to high butachlor toxicity, total sperm count dropped to 16.92 ± 2.5 billion spermatozoa/cc in 2-3 year olds and to 15.98 ± 2.5 billion spermatozoa/cc in fishes above 3 years of age. Similarly total sperm count in R. frisii kutum exposed to low butachlor toxicity was recorded as 23.6 ± 2.5 billion spermatozoa/cc in 2-3 year olds and 29.4 ± 2.5 billion spermatozoa/cc in fishes above 3 years of age. Under normal conditions, on the basis of morphology, spermatozoa showed only 10 ± 1.92% of abnormal sperms. The number of abnormal sperms increased by 28.6 ± 1.92% in fishes exposed to high butachlor toxicity, while that in fishes exposed to low butachlor toxicity increased by 19.7 ± 1.92% in 2-3 year olds and 16.6 ± 19.2% in fishes above 3 years of age. It is evident from the results obtained that increase in level of pollution caused a decrease in sperm volume but an increase in the percentage of abnormal sperms. Results obtained indicate that exposure to high butachlor toxicity (75% of its LC50 value) decreased testostron hormone to 0.31 ± 0.22 ng/ml in high butachlor toxicity, and to 0.45 ± 0.22 ng/ml in low butachlor toxicity (25% of its LC50 value). Testostron hormone dropped to 0.53 ± 0.22 ng/ml in 2-3 year olds and to 0.79 ± 0.22ng/ in fishes above 3 years of age. The testostron hormone under normal conditions in R. frisii kutum is 2.7 ± 0.22 ng/ml. It is evident from the results obtained that increase in level of pollution caused a decrease in testostron hormone

Changes of tissue endothelin-1 and nitric oxide synthase in a sheep model of large intestinal obstruction

Large intestinal obstruction (LIO) in farm animals can cause a ischaemic necrosis of intestinal tissue, eventually leading to death. The roles of endothelin-1 (ET-1) and nitric oxide (NO) are not well understood in the process of LIO, but evidence suggests that endothelial-derived mediators may participate. In the present study, ET-1 concentration and total nitric oxide synthase (NOS) activity were measured in heart, liver, pancreas, lung and kidney in a model of LIO in sheep. Our data demonstrated that ET-1 concentration and NOS activity were altered, with significant increases of ET-1 in heart, lung and kidney and of NOS activity in pancreas and kidney, but a marked decline of NOS activity in liver (p<0.05). It is postulated that these alterations in NOS activity and ET-1 concentration may contribute to the progressive loss of organ function, and finally lead to death in LIO in sheep.

The effect of artisanal preservation methods on nutritional security of “Mukene” Rastrineobola argentea caught from Lakes Victoria and Kyoga in Uganda

The artisanal fish preservation methods in Uganda are characterized by extreme operating conditions. Consequently, vital nutritional components diminish in value and quantity which renders fish consumer nutritionally insecure. To establish the magnitude of nutritional loss, duplicate samples of Mukene Rastrineobola argentea were collected from Kiyindi landing site on L. Victoria and Moone landing site on L. Kyoga. Each set of duplicate samples was divided into five portions and kept on ice. For each preservation method a portion was processed into respective products at Food Bioscience and Agri-Business Laboratories aside from the control (fresh) sample. Both preserved and control samples were analysed for nutrient loss at Department of Chemistry, Makerere University using AOAC methods. The composition of fatty acids was determined by methanolysis gas chromatography and Mass spectrophotometry of the resultant methyl esters. The results indicate that nutrients of all preserved samples did not vary significantly from the control except for some fatty acids. The Eicosapentaenoic acid (EPA) in fresh samples declined from 6.72% to 1.08% in deep-fried samples constituting 83.93% nutrient loss. The sum ratio w3:w6 as well as EPA: DHA (Docosahexaenoic) ratio in fried samples also varied significantly (p<0.5) lower than 0.668 and 0.20 for the average of either preservation methods and experts recommended ratio respectively. Further research has been recommended to ascertain the causative factor, since Mukene frying is being promoted in the Great lakes region as alternative method to sun-drying. In conclusion, regular consumers of fried Mukene do not benefit much from the nutritional and health attributes of Omega 3 and 6.

The preservation of some East African freshwater fish

Quality changes during storage were investigated for several commercially important East African freshwater fish. Lates, Bagrus, Protopterus, Tilapia esculenta and T. nilotica were examined during storage in ice and at ambient temperature (250•C). After 24 hours at ambient temperature Lates and Bagrus were completely spoilt but Protopterus was still edible. In iced storage most fish were acceptable for at least 20 days. Organoleptic examination showed that T. nilotica was acceptable after 22 days storage in ice and that gutting was only marginally beneficial. Changes in physical appearance, which could form the basis of a fish inspection system, were recorded during storage. Possible chemical quality control indices were also investigated. It was found that total volatile bases and hypoxanthine are unlikely to be useful quality indices for the species studied with the possible exception of Lates. The bacterial counts of the flesh and skin of T. esculenta and T. niloticus were found to be low (a maximum of 10 organisms per sq cm of skin or per g of flesh) after 22 days storage in ice.

HORMONAL-REGULATION OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR AND PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-1 IN CULTURED MONKEY SERTOLI CELLS

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5x10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 mu g/cm(2)) in 0.5 mi McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 mi serum-free medium with or without various hormones or other compounds, PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed, Monkey Sertoli cells were also capable of secreting PAI-1, Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity. PAI-1 activity in the culture was also enhanced by these reagents except 8-bromo-dibutyryl-cAMP, forskolin and 3-isobutyl-1-methylxanthin (MIX) which greatly stimulated tPA activity, whereas decreased PAI-1 activity, implying that neutralization of PAI-1 activity by tile high level of tPA in the conditioned media may occur. These data suggest that increased intracellular signals which activate protein kinase A (PKA), or protein kinase C (PKC) can modulate Sertoli cell tPA and PAI-1 activities, The concomitant induction of PA and PAI-1 by the same reagents in the Sertoli cells may reflect a finely tuned regulatory mechanism in which PAI-1 could limit the excession of the proteolysis.

Localization and distribution of tissue type and urokinase type plasminogen activators and their inhibitors type 1 and 2 in human and rhesus monkey fetal membranes

Fetal membranes consist of 10 distinct layers including components of amnion, chorion and decidua, the latter being of maternal origin. They form mechanically integrated sheets capable of retaining amniotic fluid and play an essential role in protecting fetal growth and development in the pregnant uterus. The extracellular matrix, substrate for plasminogen activators (PAs), is an important supportive framework of the fetal membranes. :Fetal membranes from women with preterm premature rupture of membranes may differ in their protease activity compared with normal membranes. To identify the presence of PAs and their inhibitors (PAI) and their possible role in the process of fetal membrane rupture, this study in investigated the distribution and localization of both protein and mRNA for tissue (t) and urokinase (u) PA and their inhibitors type 1 (PAI-1) and type 2 (PAI-2) in amniochorion of human and rhesus monkey using conventional and. confocal immunofluorescence microscopy. In situ hybridization analysis showed that the distribution and localization of mRNAs for tPA, uPA, PAI-I and PAI-2 were similar in the fetal membranes of human and rhesus monkey; no obvious species difference was observed. Evidence of tPA mRNA was detected in amniotic epithelium, trophoblast cells and nearly all cells of the decidual layer. Strong expression of uPA mRNA was noted in the decidual cells which increased in intensity as the abscission point was approached. Weak staining in chorion laeve trophoblast was also detected. In situ hybridization experiments showed PAI-1 mRNA to be concentrated mainly in the decidual cells, some of which were interposed into the maternal-facing edge of the chorion laeve. Maximal labelling of the decidua occurred towards the zone of abscission. Weak expression of PAI-1 mRNA nas also noted in some cells of the chorion laeve. The distribution of PAI-2 mRNA in amniochorion was also concentrated in the cells of the decidual layer, maximum expression of the mRNA was in the level of abscission. No detectable amount of mRNAs for tPA, uPA, PAI-1 and PAI-2 was found in the fibroblast, reticular and spongy layers. Distribution of the proteins of tPA, uPA and PAI-1 in the fetal membranes of these two species was consistent with the distribution of their mRNA. Anti-PAI-2 immunofluorescence was found to be strongly concentrated in the amniotic epithelium, but PAI-2 mRNA was negative in this layer, suggesting that the epithelium-associated PAI-2 is not of epithelial origin. These findings suggest that a local fibrinolysis in fetal membranes generated by precisely balanced expression of PAs and their inhibitors via paracrine or autocrine mechanisms may play an essential role in fetal membrane development, maturation and in membrane rupture. Following an analysis of the distribution and synthesis of activators and inhibitors it was found that they may play a role in abscission during the third stage of labour. (C) 1998 W. B. Saunders Company Ltd.

Influence of leaf tissue structure on host feeding selection by pea leaf miner Liriomyza huidobrensis (Diptera : Agromyzidae)

Host feeding selection by the female pea leafminer, Liriomyza huidobrensis, on 47 species of plants was studied. The leaves were sectioned by microtome, and 15 characteristics of the leaf tissue structure were measured under a microscope. Correlation analysis between host feeding selection and leaf tissue structure indicated that the preference of host feeding selection was positively correlated with the percentage of moisture content of leaves and negatively with thickness of the epidermis wall, and densities of the palisade and spongy tissues of leaves. Leaf tissue structure was influential in feeding and probing behavior of female L. huidobrensis. So, thickness of epidermis wall, densities of the palisade and spongy tissues can act as a physical barrier to female oviposition. Furthermore, higher densities of palisade and spongy tissues can be considered a resistant trait which affects mining of leaf miner larvae as well. As a result, plants with lower leaf moisture content may not be suitable for the development of L. huidobrensis.

Composite hydrogels for nucleus pulposus tissue engineering

Tissue engineering offers a paradigm shift in the treatment of back pain. Engineered intervertebral discs could replace degenerated tissue and overcome the limitations of current treatments, which substantially alter the biomechanical properties of the spine. The centre of the disc, the nucleus pulposus, is an amorphous gel with a large bound water content and it can resist substantial compressive loads. Due to similarities in their compositions, hydrogels have frequently been considered as substitutes for the nucleus pulposus. However, there has been limited work characterising the time-dependent mechanical behaviour of hydrogel scaffolds for nucleus pulposus tissue engineering. Poroelastic behaviour, which plays a key role in nutrient transport, is of particular importance. Here, we investigate the time-dependent mechanical properties of gelatin and agar hydrogels and of gelatin-agar composites. The time-dependent properties of these hydrogels are explored using viscoelastic and poroelastic frameworks. Several gel formulations demonstrate comparable equilibrium elastic behaviour to the nucleus pulposus under unconfined compression, but permeability values that are much greater than those of the native tissue. A range of time-dependent responses are observed in the composite gels examined, presenting the opportunity for targeted design of custom hydrogels with combinations of mechanical properties optimized for tissue engineering applications. © 2011 Elsevier Ltd.