Evaluation of Potential Thrombin Inhibitors from the White Mangrove (Laguncularia racemosa (L.) C.F. Gaertn.)

The aim of this work was to verify the effects of methanol (MeOH) and hydroalcoholic (HA) extracts and their respective partition phases obtained from white mangrove (Laguncularia racemosa (L.) C.F. Gaertn.) leaves on human thrombin activity. Among the extracts and phases tested, only the ethyl acetate and butanolic partitions significantly inhibited human thrombin activity and the coagulation of plasma in the presence of this enzyme. Chromatographic analyses of the thrombin samples incubated with these phases revealed that different compounds were able to interact with thrombin. The butanolic phase of the MeOH extract had the most potent inhibitory effects, reducing enzymatic activity and thrombin-induced plasma coagulation. Two glycosylated flavonoids in this partition were identified as the most potent inhibitors of human thrombin activity, namely quercetin-3-O-arabinoside (QAra) and quercetin-3-O-rhamnoside (Qn). Chromatographic analyses of thrombin samples incubated with these flavonoids demonstrated the chemical modification of this enzyme, suggesting that the MeOH extract contained other compounds that both induced structural changes in thrombin and diminished its activity. In this article, we show that despite the near absence of the medical use of mangrove compounds, this plant contains natural compounds with potential therapeutic applications.     

Testing the relationship between instantaneous peak flow and mean daily flow in a Mediterranean Area Southeast Spain

Extreme hydrologic events have great importance in Southeastern Spain regarding the personal and economic damage they imply. The commonly-used design parameter for hydraulic structures is the maximum annual instantaneous stream flow recorded in conventional gauging stations. However, the majority of available data in Southeastern Spain is mean daily stream flows. This paper explores possible linear relationships between annual instantaneous peak discharge (IPF) and the corresponding (MDF) mean daily stream flow. This relationship was previously explored by other authors such as Fuller [Fuller, W.E., 1914. Flood flows. Trans. Am. Soc. Civ. Eng., 77: 564–617]. Non-linear responses of IPF–-MDF were observed in several study basins. The use of Principal Components Analysis (PCA) allowed characterizing the most important topographic and hydrological attributes of the basins and provided important information about variables that should be included in IPF–MDF regional equations. The key factor to justify the different IPF–MDF relationships in a relatively small area is the nature of the extreme events and their effects on semi-arid soil conditions. In addition, a regional equation to estimate IPF from MDF was developed. This equation was applied to a series of flow of nine stations of the Southeast Basin of Spain, and a significant improvement was achieved when applying this formula in comparison to the traditional method of Fuller. This study indicates possible restrictions to take into account when traditional hydrological model are applied in semi-arid areas.     

Lectin Histochemistry as a Tool to Identify Apoptotic Cells in the Seminiferous Epithelium of Syrian Hamster (Mesocricetus auratus) Subjected to Short Photoperiod

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con‐A, UEA‐I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal β1,3‐GalNAcα1, α‐d ‐mannose, N‐acetylgalactosamine and l ‐fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.     

Idiopathic eosinophilic meningoencephalitis in Rottweiler dogs: three cases (1992–1997)

We report three cases of eosinophilic meningitis in young male Rottweiler type dogs in New Zealand. No cause for the disease was identified. There were variable clinical signs referable to central nervous system dysfunction, and a variable response to treatment.     

Protein degradation by rumen microbes of heat-treated whole cottonseed

In vitro and in situ rumen digestion trials were conducted to measure the effect of heat treatment on protein degradation by rumen microbes of cottonseed autoclaved and dry-heated (120, 140, 160 and 180 C) for different lengths of time (20, 40, 60, 90 and 120 min). Autoclaving (120 C, 1 kg/cm2 of pressure for 60 min) was effective in reducing ammonia nitrogen concentration in vitro, but dry heat at this temperature was without effect. Dry heating caused a linear decrease of in vitro ammonia concentration as temperature increased from 140 to 180 C and as exposure time increased from 20 to 120 min. The in situ results corroborated the in vitro findings. Crude protein disappearance from dacron bags decreased from 87 to 48% as temperature increased from 140 to 180 C after 20 min of heating.     

Redox cycling induces spermptosis and necrosis in stallion spermatozoa while the hydroxyl radical (OH•) only induces spermptosis

Oxidative stress is a major factor explaining sperm dysfunction of spermatozoa surviving freezing and thawing and is also considered a major inducer of a special form of apoptosis, visible after thawing, in cryopreserved spermatozoa. To obtain further insights into the link between oxidative stress and the induction of apoptotic changes, stallion spermatozoa were induced to oxidative stress through redox cycling after exposure to 2‐methyl‐1,4‐naphthoquinone (menadione), or hydroxyl radical formation after FeSO₄ exposure. Either exposure induced significant increases (p < 0.05) in two markers of lipid peroxidation: 8‐iso‐PGF_2α and 4‐hydroxynonenal (4‐HNE). While both treatments induced changes indicative of spermptosis (caspase‐3 activation and decreased mitochondrial membrane potential) (p < 0.01), menadione induced sperm necrosis and a dramatic reduction in motility and thiol content in stallion spermatozoa. Thus, we provided evidence that oxidative stress underlies spermptosis, and thiol content is a key factor for stallion sperm function.     

Changes in Testicular Interstitial Connective Tissue of Hamsters (Mesocricetus auratus) During Ageing and After Exposure to Short Photoperiod

The testicular interstitium of Syrian hamster (Mesocricetus auratus) was studied during ageing and in testicular regression after exposure to a short photoperiod, in relation to the interstitial cells and their connective tissue. This tissue was assessed histochemically using Masson's trichrome technique and the expression of Heat Shock Protein 47 (HSP‐47) and collagen IV (α5) was assessed in Leydig cells. Finally, an ultrastructural analysis of some cells of the testicular interstitium was made. Leydig cells were positive for HSP‐47 and collagen IV (α5). Ageing did not change the parameters studied while the short photoperiod altered the synthetic activity of Leydig cells. The positivity index of these cells for HSP‐47 was significantly higher in the regressed testis, but was lower for collagen IV (α5). During ageing no change were observed. Ultrastructural Leydig cells showed a discontinuous basal lamina that did not change during ageing. The basal lamina was not identified in Leydig cells regressed by exposure to a short photoperiod. In conclusion; the intertubular connective tissue suffers little change with age. By contrast, in the testis regressed after exposure to a short photoperiod the studied parameters related to the intertubular connective tissue were altered. These changes are probably related with the low synthetic activity of regressed Leydig cell.     

Soil genesis and heterogeneity of phosphorus forms and carbon below mounds inhabited by primary and secondary termites

Termite activities are known to significantly influence small-scale soil properties in tropical savannas. The lateral and vertical extent of the alterations to the nest's surrounding, and particularly resulting impacts on diagnostic soil horizons remain largely unresolved until today. We examined the effects of mound-building termites on soil genesis and constitutive chemical soil properties in and below their nests. Two transects to a soil depth of 100 cm were dug below three younger mounds of Cornitermes silvestrii (the primary nest builder), three older mounds in which C. silvestrii had died out and which were secondarily colonized mainly by Nasutitermes kemneri, and three reference sites in the Brazilian Cerrado. The samples were characterized by standard procedures for soil classification; in addition, phosphorus extractions were conducted on selected samples using NaHCO₃ for labile P forms, and concentrated HCl for stable P forms. This data set was then used to build calibration models for the prediction of labile and stable inorganic (P_i) and organic (P_o) P forms, as well as for contents of organic carbon (OC), for the remaining samples applying mid-infrared spectroscopy in combination with partial least squares regression (MIRS-PLSR). We can show that the termite influence on the soil was sufficiently large to change diagnostic characteristics of the soils under termite mounds. The MIRS-PLSR predictions were suitable for quantifying organic carbon and most of the labile and stable phosphorus fractions. They showed an enrichment of OC, NaHCO₃-P_o and NaHCO₃-P_i contents in nests inhabited by primary and secondary termites by factors of 1.6–2.0 and 1.4–1.5, respectively. The soils surrounding the nests had higher contents of OC and NaHCO₃-P under both nest types vertically down to 30 cm below the lower nest border, and OC and NaHCO₃-P_i contents were elevated at minimum to a lateral distance of 60 cm away from the nest border. As the pattern of HCl_conc-P_i, which comprised 95% of total P, showed no variations, we conclude that the higher NaHCO₃-P_i amount was formed in termite nests by changing the availability of the more stable HCl_conc-P_i. In contrast to the contents, the OC and NaHCO₃-P stocks below the mounds inhabited by primary termites were comparable to those inhabited by secondary ones, because the bulk density of the secondarily inhabited nests was elevated. This was due to a transport of clay-rich material from the subsurface argic horizons into the nests. Here, the secondary termites even reverted the lessivation observed in the reference soils and under mounds inhabited by primary termites, thus causing the soil types to change from Alisols and Acrisols to the properties of Umbrisols.     

DECR1 and ME1 genotypes are associated with lipid composition traits in Duroc pigs

Variation at the porcine DECR1 and ME1 genes has been associated with meat quality traits and backfat thickness in Landrace pigs, respectively. However, it has not been investigated yet whether DECR1 and ME1 genotypes influence lipid composition. With this aim, we have genotyped two missense DECR1 substitutions (c.160G>C and c.437G>C) and one silent ME1 (c.576C>T) polymorphism in 361 Duroc barrows distributed in five half‐sib families and phenotyped for serum lipid concentrations and intramuscular fat content and composition traits. At the whole‐population level, relevant associations, that is, with a posterior probability of the allele substitution effect to be over or below zero (PPN0) > 0.90, were observed between DECR1 genotype and serum cholesterol (CHOL) (PPN0 = 0.932) and LDL concentrations (PPN0 = 0.945) at 190 days, as well as between ME1 genotype and longissimus dorsi saturated fatty acid content (PPN0 = 0.924). At the within‐family level, we found relevant associations between DECR1 and ME1 genotypes and diverse lipid composition traits, but most of them were family‐specific. Discrepancies in allele substitution effects estimated in half‐sib families might be produced by many factors such as number of individuals, marker allele frequencies and informativeness in each family, unaccounted random genetic and environmental effects, epistasis and family‐specific differences in the linkage phase or amount of linkage disequilibrium between causal and marker mutations. This lack of consistency across families, combined with the fact that the ME1 mutation is synonymous and that the two DECR1 polymorphisms are conservative, suggests that the associations found are not causative.     

Mitochondria in Mammalian Sperm Physiology and Pathology: A Review

While, for a long time, the role of mitochondria in sperm physiology and pathology has been largely ignored, recent research points out the mitochondria as a major organelle with key roles in sperm function both under physiological and biotechnological conditions. This paper briefly reviews these novel findings regarding the role of mitochondria in sperm, paying special attention to the most practical, readily applicable, aspects of the topic such as their role as a major source of the sublethal damage that sperm experiments after cryopreservation.