Transplantation of human fetal blood stem cells in the osteogenesis imperfecta mouse leads to improvement in multiscale tissue properties.

Osteogenesis imperfecta (OI or brittle bone disease) is a disorder of connective tissues caused by mutations in the collagen genes. We previously showed that intrauterine transplantation of human blood fetal stem/stromal cells in OI mice (oim) resulted in a significant reduction of bone fracture. This work examines the cellular mechanisms and mechanical bone modifications underlying these therapeutic effects, particularly examining the direct effects of donor collagen expression on bone material properties. In this study, we found an 84% reduction in femoral fractures in transplanted oim mice. Fetal blood stem/stromal cells engrafted in bones, differentiated into mature osteoblasts, expressed osteocalcin, and produced COL1a2 protein, which is absent in oim mice. The presence of normal collagen decreased hydroxyproline content in bones, altered the apatite crystal structure, increased the bone matrix stiffness, and reduced bone brittleness. In conclusion, expression of normal collagen from mature osteoblast of donor origin significantly decreased bone brittleness by improving the mechanical integrity of the bone at the molecular, tissue, and whole bone levels.

Effects of blood extraction on horseshoe crabs (Limulus polyphemus)

Horseshoe crabs (Limulus polyphemus) are caught by commercial fishermen for use as bait in eel and whelk fisheries (Berkson and Shuster, 1999)—fisheries with an annual economic value of $13 to $17 million (Manion et al.1). Horse-shoe crabs are ecologically important, as well (Walls et al., 2002). Migratory shorebirds rely on horseshoe crab eggs for food as they journey from South American wintering grounds to Arctic breeding grounds (Clark, 1996). Horse-shoe crabs are also essential for public health (Berkson and Shuster, 1999). Biomedical companies bleed horse-shoe crabs to extract a chemical used to detect the presence of endotoxins pathogenic to humans in injectable and implantable medical devices (Novitsky, 1984; Mikkelsen, 1988). Bled horseshoe crabs are returned to the wild, subject to the possibility of postbleeding mortality. Recent concerns of overharvesting have led to conflicts among commercial fishermen, environmentalists acting on behalf of the shorebirds, and biomedical companies (Berkson and Shuster, 1999; Walls et al., 2002).

Acute effect of sumithion on certain blood parameters and mortality rate of fish, Heteropneustes fossilis (Bloch)

The acute toxic effect of the toxicant sumithion (50% E.C.) on mortality rate (after 24, 48, 72, and 96 h), total RBC count and haemoglobin content (after 48 and 72 h) on Heteropneustes fossilis was investigated at four concentrations (9.7, 10.7 and 11.1 ppm). The sumithion treated fishes showed lower RBC and Hb levels than the untreated ones. A gradual decrease in the total RBC counts and Hb contents was recorded with increasing concentration of toxicant after 72 h but the blood showed fluctuating values after 48 h of treatment.

Toxic effects of dietary seaweeds Sargassum bovianum, Caulera faridii and Gracilaria corticata on blood parameters of albino rats

The present communication deals with the feeding trials of brown (Sargassum bovianum), green (Caulerpa faridii) and red (Gracilaria corticola) seaweeds in albino rats for a period of thirty days in order to investigate their digestibility and acceptability as supplementary food for animals. The parameters used were: changes in blood hemoglobin, ESR, MCHC, PCV and plasma vitamin levels. The result revealed that all the three species of seaweeds had acceptability up to 5% level, as no ill effect was noted during the experiment. But at 10% and 20% levels, marked changes were observed in blood parameters with diarrhea, vomiting and convulsions indicating possibilities of either tissue and muscular dystrophy, gastrointestinal tract necrosis or functional disorder of central nervous system. A heavy mortality was noted due to excessive water loss through diarrhea and vomiting. However, no mortality was observed after 22nd day at both 10% and 20% levels with subsided clinical signs. The results suggest that these three seaweed species could be used safely as a supplementary food, in native form, in animals at low concentrations.

Postmortem changes in hilsa fish. Pt. 2. Studies on physical and bacteriological changes in iced and frozen stored hilsa fish (Tenualosa ilisha Ham.)

Studies were conducted to evaluate the quality of hilsa fish during icing and freezing storage at -20°C by determining organoleptic and bacteriological aspects. The fishes stored in ice were organoleptically in acceptable condition 2 for 20 days. The bacterial load in muscles of 4 days ice stored fish was 2.5x10² CFU/g which gradually increased up to 1.8x10⁵ CFU/g after 20 days when the fishes were organoleptically in acceptable condition. The keeping qualities of different days of ice stored fishes were also evaluated during their subsequent frozen storage at -20°C. Both 4 and 7 days of ice stored fishes were organoleptically in acceptable condition up to 48 weeks but the highest degree of freshness was found for fish stored in ice for 4 days before freezing at -20°C. The result indicates that the longer is the duration of ice storage before freezing, the shorter is the shelf life of the fish. The initial bacterial load prior to freezing of the 4 and 7 days of ice stored samples were 2.5x10³ CFU/g and 3.8x10⁴ CFU/g, respectively which reduced to 2.21x10² CFU/g and 2.38x10² CFU/g, respectively at the end of the 24 weeks of frozen storage. However, after 40 weeks the bacterial load in the frozen stored sample fell below the detection level.

Cell types in the peripheral blood of waking catfish Clarias batrachus (Lin.)

Different types of haematocytes found in the peripheral blood of walking catfish Clarias batrachus, have been characterized and identified using morphological, morphometric and cytochemical techniques. These cells are: erythrocytes, reticulocytes, large and small lymphocytes, thrombocytes, monocytes and polymorphonuclear leucocytes (neutrophils).

Suitability of ice stored mackerel and sardine for canning

The paper reports the results of studies on ice storage and subsequent canning of mackerel (Rastrelliger kanagurta) and Sardines (Sardinella longiceps) and the effect of such storage on the quality of the canned product prepared out of them. The changes in the physical and chemical characteristics during ice storage are determined and correlated with the quality of the finished product.

Sulfide binding characteristics of blood serum in Calyptogena pacifica and Vesicomya gigas

The sulfide binding characteristics of blood serum were studied in vitro in two deep-sea vesicomyid clams, Calyptogena pacifica and Vesicomya gigas. Both the C. pacifica and the V. gigas serum concentrated sulfide at least an order of magnitude above ambient levels. V. gigas accumulated sulfide faster than C. pacifica, reaching saturation at 5000 M after an hour. C. pacifica bound sulfide at half the rate of V. gigas, reaching saturation in about two hours at a substantially higher concentration of sulfide. The observed distribution of the animals near cold seeps in the Monterey Submarine Canyon can be explained by their different sulfide binding abilities. The hypothesis that cold seeps are actually much more unstable sources of sulfide than previously assumed is explored.

Genetic diversity of Yunnan local pig breeds inferred from blood protein electrophoresis

Protein electrophoresis was used to examine the blood protein polymorphism in Yunnan local pig breeds, i.e., the Saba pig, Dahe pig, and Diannan small-ear pig breeds, Of 38 genetic loci surveyed 9 were found to be polymorphic. The percentage of polymorphic loci (P) varies from 0.1875 to 0.2121, and the mean individual heterozygosity (H) varies front 0.0712 to 0.1027 in three pig breeds. The results indicate that blood protein polymorphism in Yunnan pig breeds is high. Yunnan local pig breeds have a wealth of genetic diversity at the level of blood proteins.

In vitro phagocytic study of blood leucocytes and peritoneal macrophages of walking catfish Clarias batrachus ( Ham.) against Aeromonas hydrophila and Escherichia coli.

In observation of in vitro phagocytic activity against Aeromonas hydrophila isolate 34k (a virulent form) and Escherichia coli (an avirulent bacteria) of neutrophil- and monocyte-like cells of walking catfish Clarias batrachus showed phagocytosis. N eutrophils and monocytes phagocytized the avirulent form of bacterial isolate more than the virulent one. Other blood leucocytes did not show phagocytosis. Peritoneal macrophage of the fish were separated by glycogen elicitation and the macrophages were being adhered on plastic cover slips for studying their in vitro phagocytic activity. Most of the cells were alive after adherence and showed phagocytosis against the virulent and avirulent bacteria. The percent phagocytosis and phagocytic index were higher against the avirulent E. coli than the virulent A. hydrophila.

ISOLATION AND PROPERTIES OF A BLOOD-COAGULATION FACTOR-X ACTIVATOR FROM THE VENOM OF KING COBRA (OPHIOPHAGUS HANNAH)

A specific blood coagulation factor X activator was purified from the venom of Ophiophagus hannah by gel filtration and two steps of FPLC Mono-Q column ion-exchange chromatography. It showed a single protein band both in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and alkaline polyacrylamide gel electrophoresis. The mol. wt was estimated to be 62,000 in non-reducing conditions and 64,500 in reducing conditions by SDS-PAGE. The isoelectric point was found to be pH 5.6. The enzyme had weak amidolytic activities toward CBS 65-25, but it showed no activities on S-2266, S-2302, thrombin substrate S-2238, plasmin substrate S-2251 or factor Xa substrate S-2222. It had no arginine esterase activity toward substrate benzoylarginine ethylester (BAEE). The enzyme activated factor X in vitro and the effect was absolutely Ca2+ dependent, with a Hill coefficient of 6.83. It could not activate prothrombin nor had any effect on fibrinogen and thus appeared to act specifically on factor X. The procoagulant activity of the enzyme was almost completely inhibited by serine protease inhibitors like PMSF, TPCK and soybean trypsin inhibitor; partially inhibited by L-cysteine. Metal chelator EDTA did not inhibit its procoagulant activity. These results suggest that the factor X activator from O. hannah venom is a serine protease.

AN ACTIVATOR OF BLOOD-COAGULATION FACTOR-X FROM THE VENOM OF BUNGARUS-FASCIATUS

A specific activator of blood coagulation factor X was purified from the venom of Bungarus fasciatus by gel filtration and by ion-exchange chromatography on a Mono-Q column (FPLC). It consisted of a single polypeptide chain, with a mel. wt of 70,000 in reducing and non-reducing conditions. The enzyme had an amidolytic activity towards the chromogenic substrates S-2266 and S-2302 but it did not hydrolyse S-2238, S2251 or S-2222, which are specific substrates for thrombin, plasmin and factor Xa, respectively. The enzyme activated factor X in vitro and the effect was Ca2+ dependent with a Hill coefficient of 7.9. As with physiological activators, the venom activator cleaves the heavy chain of factor X, producing the activated factor Xa alpha. The purified factor X activator from B. fasciatus venom did not activate prothrombin, nor did it cleave or clot purified fibrinogen. The amidolytic activity and the factor X activation activity of the factor X activator from B. fasciatus venom were readily inhibited by serine protease inhibitors such as diisopropyl fluorophosphate (DFP), phenylmethanesulfonyl fluoride (PMSF), benzamidine and by soybean trypsin inhibitor but not by EDTA. These observations suggest that the factor X activator from B. fasciatus venom is a serine protease. It therefore differs from those of activators obtained from Vipera russelli and Bothrops atrox venoms, which are metalloproteinases.

Effects of Pallas' viper (Agkistrodon halys pallas) venom on blood coagulation and characterization of a prothrombin activator

The action of Pallas' viper (Agkistrodon halys pallas) venom on blood coagulation was examined in vitro and a strong anticoagulant effect was observed. This action was abolished after treatment with a specific inhibitor of phospholipase A(2) activity (p-bromophenacyl bromide), revealing a procoagulant action in low concentrations of treated venom (around 1 mu g/ml). The effect of the venom an haemostasis was further characterized by measuring its ability to activate purified blood coagulation factors. It is concluded that A. halys pallas venom contains prothrombin activation activity. A prothrombin activator (aharin) was purified from the venom by Sephadex G-75 gel filtration and ion-exchange chromatography on a Mono-Q column. It consisted of a single polypeptide chain, with a mol. wt of 63,000. Purified aharin possessed no amidolytic activity on chromogenic substrates. It did not act on other blood coagulation factors, such as factor X and plasminogen, nor did it cleave or clot purified fibrinogen. The prothrombin activation activity of aharin was readily inhibited by ethylenediamine tetracetic acid (a metal chelator), but specific serine protease inhibitors such as diisopropyl fluorophosphate and phenylmethanesulfonyl fluoride had no effect on it. These observations suggest that, like those prothrombin activators from Echis carinatus and Bothrops atrox venoms, the prothrombin activator from A. halys pallas venom is a metalloproteinase. (C) 1998 Elsevier Science Ltd. All rights reserved.