Ultrastructural observations on the gills of polychaetes

The gills of several polychaete species belonging to 9 families were studied by scanning and transmission electron microscopy. The surface epithelium is covered by a thin cuticle which is invaded by microvilli penetrating the epicuticle in certain species. Some epithelial cells bear cilia, others are mucus-producing cells. The ciliary cells may be arranged in rows and maintain a constant flow of water over the gills. The distance between external water and blood stream differs considerably according to the species investigated. InMalacoceros the gills are characterized by closed afferent and efferent subepithelial vessels, which correspond to tubular invaginations of the coelomic wall. These vessels are lined by the basement lamina of the coelothelial cells, which are of the epitheliomuscular type. The vessels are open in the gills of other polychaetes and release the blood stream into a system of spaces immediately below the epidermis (e.g. in the branchial lamellae ofPectinaria andTerebellides). In several species the blood comes into very intimate contact with the cuticle (e.g. in the gill filaments ofDendronereides), but also in these animals both are separated by a very small epidermal layer.Supported by DFG Sto 75/3-6.     

Molecular basis of low-temperature sensitivity in pig hemoglobins.

It has been generally assumed that mammals have blood with a greater temperature sensitivity than ectothermic organisms. Recent results have shown that in some species of mammals, Hb displays a value of overall oxygenation enthalpy (delta H) much less exothermic than that observed for most mammalian hemoglobins, including human adult Hb. In this respect, a very interesting case is represented by porcine blood which shows a modest effect of temperature, the temperature coefficient of its oxygen-dissociation curve being significantly lower than that of human blood. Here we report a detailed functional characterization of pig Hb, which, interpreted on the basis of the amino acid sequence of the alpha and beta chains of the molecule, sheds some light on the molecular basis of the phenomenon.     

Structural studies of an eukaryotic cambialistic superoxide dismutase purified from the mature seeds of camphor tree

Inflammation is one of the leading causes of the many pathological states associated with oxidative stress. A crucial role in the development of inflammation-induced oxidative stress is played by reactive oxidant species (ROS), which are very difficult to detect in vivo. One of the most sensitive and definitive methods in the detection of ROS is electron spin resonance, especially as used in conjunction with spin trapping. Unfortunately, the commonly used nitrone spin traps have a very low efficacy for trapping superoxide radicals, and their radical adducts are not stable. To address this deficiency, we have developed negatively charged cyclic hydroxylamines such as 1-hydroxy-4-phosphonooxy-2,2,6,6-tetramethylpiperidine (PP-H) for the detection of reactive oxidant species as a diagnostic tool for extracellular inflammation-induced oxidative stress. We used inflammation induced by a bacterial endotoxin lipopolysaccharide (LPS) as a model. ROS formation was tested in cultured macrophages, in blood and in vivo. PP-H reacts with reactive oxidant species generating the stable nitroxide radical 4-phosphonooxy-TEMPO. It was shown that a 5-h treatment of macrophages with LPS (1 microg/ml) leads to a threefold increase in superoxide formation as demonstrated using superoxide dismutase. Formation of reactive oxidant species 5 h after LPS (1 mg/kg) treatment of Fischer rats was analyzed in arterial blood; formation of reactive oxidant species in LPS-treated animals increased by a factor of 2.2 and was dependent upon the LPS dose. Diphenyleneiodonium (0.1 mM) inhibited formation of LPS-stimulated reactive oxidant species by 80%. We suggest that this test could be used as a noninvasive diagnostic tool for inflammation-induced oxidative stress.     

Annotated Checklist of the Terrestrial Gastropods of Nepal

This is the very first checklist of the terrestrial gastropods of Nepal. It includes 138 species and six subspecies, of which 22 species are endemic and four are introduced. It highlights 34 species recorded for the first time in Nepal and provides new distribution records for another 30 species.     

Electrophoretic patterns of hemoglobin and oxygen binding properties of blood of anostomidae fishes from Parana-Pardo-Grande hydrographic basin (S?o Paulo State, Brazil)

The electrophoretic patterns of hemoglobin (Hb) and the functional properties of blood of seven migratory fish species, in two different kinds of habitat during both the rainy and dry seasons, were studied. All species showed multiple hemoglobins and very similar electrophoretic patterns except for Schizodon nasutus, which showed two cathodal components. The functional properties of blood reflect those electrophoretic patterns, since all species showed very little or no Bohr effect during each situation studied. Data on Leporinus obtusidens blood (during the rainy season), from both lotic and lentic environments, showed a normal Bohr effect (phi = -0.073). Similarly, Schizodon nasutus and Leporinus friderici from a lotic environment during the dry season were compared and showed a reverse Bohr effect (phi = 0.042). In the other 14 situations, only Leporinus elongatus from a lentic environment during the rainy season showed a statistically significant difference in log P50 values from the other fishes.     

Morphine metabolism in the naturally morphine-tolerant afghan pika: a preliminary study

The afghan pika (Ochotona rufescens), a lagomorph which is naturally tolerant to the analgesic action of morphine, metabolizes morphine into morphine 3-glucuronide apparently faster than does the rabbit, another lagomorph which is however normally responsive to morphine. In the two species, following morphine administration, another unidentified component appears very soon (5 min) in pika blood plasma and much later (60 min) in rabbit blood plasma. This unknown component which appears not to be morphine derived might be involved in the natural resistance of the Afghan pika to morphine.     

Blood Protozoa of Imported Birds

SYNOPSIS. Large numbers of birds, until recently, were brought into the United States each year. Countries of origin were varied, and included those of Australasia, Africa, South America, and the Caribbean Islands, as well as other places. With them of course come their parasites, some of which may be potential pathogens to domestic avifauna. In part for this reason, a survey was undertaken of blood parasites of birds from pet shops and importers. So far a total of 1234 birds belonging to 186 species has been examined. Several new species and subspecies of avian Plasmodium have been found in the course of this study, including P. octamerium Manwell, 1968 in a Pintail Whydah, Vidua macoura , from Africa; P. paranucleophilum Manwell & Sessler, 1971 in a South American tanager, Tachyphonus sp.; and P. nucleophilum toucani Manwell & Sessler 1971 in a Swainson's Toucan, Ramphastos s. swainsonii. Plasmodium huffi Muniz, Soares & Battista is undoubtedly a synonym pro parte for the last. Plasmodium tenue Laveran & Marullaz, long thought to be a synonym of Plasmodium vaughani Novy & MacNeal, was rediscovered and found to be a valid species. Plasmodium nucleophilum , infrequently seen in the New World, occurred in many Asian and African birds, and especially in starlings. Infections with other species of Plasmodium were common. Haemoproteus was the commonest blood parasite; Leucocytozoon was very rare as was Atoxoplasma (Lankesterella). The 2 families of birds best represented were the Fringillidae and the Psittacidae, but no blood parasites were seen in the latter. It is clear that imported birds are often infected with blood protozoa, some of which are unknown from native birds.     

Phenomenon of indirect isoserological incompatibility in experimental transfusion research and its clinical significance

It has been established in experiments on dogs exposed to blood transfusions that if the animals are screened according to the cross-matching testing system and show as a result the lack of red blood cell agglutination, this does not provide evidence in favour of their complete isoserological compatibility. To confirm whether the animals are compatible, it is required to carry out 7 to 8 tests with other species. If the tests reveal red blood cell agglutination, blood transfusion is characterized by demonstrable isoserological incompatibility and the recipient dies. If the cross-matching with the other animals of the group is accompanied by red blood cell agglutination but the latter is absent in the testing of a particular animal pair, such a condition is designated as "indirect" incompatibility, since hemotransfusion in this animal pair is characterized by the manifestations of incompatibility. However, it is not so pronounced and the recipient can be saved by intensive care. The authors conclude that isoserological incompatibility has different grades of intensity and offer methods for the screening of animals for simulation of graver and facilitated grades of incompatibility. It is assumed that the clinical transfusiology also has grades of the severity of hemotransfusion-induced conflicts, particularly in cardiosurgery. The use of large amounts of donor's blood for extracorporeal circulation and making for large blood losses is very likely to entail isoserological incompatibility cases which may remain unrecognized because of the gravity of surgical intervention, provoking at the same time the postoperative acute heart failure.     

Respiratory Adaptations in Marine Mammals

This paper is a discussion of some of the possible structuraland functional modifications of the lung which represent adaptationsin mammals living in the sea. Lung capacities of marine mammalsseem to be larger than terrestrial mammals especially if theyare compared on a lean weight basis. It is proposed that atleast in some this represents an important increase in buoyancywhich enables these mammals to rest at sea. The importance ofthe lung as an O₂ store during dives is considered, and it seemsthat it would be important only to those species that have alow breath-hold tolerance. In their case the O₂ present in thefully inflated lung is from four times to equal that in theblood. In those species with a large breath-hold tolerance thelung O₂ store is a small fraction of blood stores. Several experimentsare discussed which indicate that during dives to depth gasexchange between the blood and lungs is low. One of the reasonssuggested is compression collapse of the alveoli. This occursbecause of the apparent rigidity of the airways which even inthe terminal segments possess an unusual amount of muscularor cartilaginous support. The reinforcement insures that duringcompression the airways will not occlude and trap gas in thealveoli. In fact, in some species, especially otariids and cetaceans,the airways seem overly strong and an additional function issuggested. Studies of mechanical properties of sea lion andwhale lungs show that they may be capable of high expiratoryflow rates at low volumes. This feature of the lung would makepossible an exchange of a large gas volume in very short periods.Such an ability is consistent with the ventilatory behaviorof many marine mammals.     

Comparison of various approaches to calculating the optimal hematocrit in vertebrates

An interesting problem in hemorheology is to calculate that volume fraction of erythrocytes (hematocrit) that is optimal for transporting a maximum amount of oxygen. If the hematocrit is too low, too few erythrocytes are present to transport oxygen. If it is too high, the blood is very viscous and cannot flow quickly, so that oxygen supply to the tissues is again reduced. These considerations are very important, since oxygen transport is an important factor for physical performance. Here, we derive theoretical optimal values of hematocrit in vertebrates and collect, from the literature, experimentally observed values for 57 animal species. It is an interesting question whether optimal hematocrit theory allows one to calculate hematocrit values that are in agreement with the observed values in various vertebrate species. For this, we first briefly review previous approaches in that theory. Then we check which empirical or theoretically derived formulas describing the dependence of viscosity on concentration in a suspension lead to the best agreement between the theoretical and observed values. We consider both spatially homogeneous and heterogeneous distributions of erythrocytes in the blood and also possible extensions, like the influence of defective erythrocytes and cases where some substances are transported in the plasma. By discussing the results, we critically assess the power and limitations of optimal hematocrit theory. One of our goals is to provide a systematic overview of different approaches in optimal hematocrit theory.     

Plasmodium paranucleophilum n. sp. from a South American Tanager*

SYNOPSIS. The avian malaria parasite described in this paper was isolated from a South American tanager of uncertain species, somewhat tentatively identified as belonging to the genus Tachyphonus. It is believed to have come from northern Brazil. Because the erythrocytic stages are, except for minor differences, similar to those of Plasmodium nucleophilum it is proposed to call it P. paranucleophilum. The chief difference separating the 2 species is the appearance of phanerozoites of the new species in lymphocytes of the circulating blood; these are characteristically vacuolated and give rise to moderate numbers of merozoites, probably in most cases 50 or less. Strains of P. nucleophilum isolated from other passerine birds also produce phanerozoites parasitizing lymphocytes, but such cells have never been observed in the blood stream. A subspecies, P. nucleophilum toucani, which we isolated from a Swainson's toucan Ramphastos swainsonii, also produces phanerozoites invading lymphocytes but it is less selective; endothelial cells of the brain are heavily attacked. It is also very lethal to canaries, whereas passerine strains of P. nucleophilum are usually quite benign. P. paranucleophilum, if the strain we have isolated is typical, lies between the 2 in pathogenicity, causing marked anemia but seldom death.     

2-DE-based proteomic investigation of the saliva of the Amazonian triatomine vectors of Chagas disease: Rhodnius brethesi and Rhodnius robustus

The triatomine bugs are obligatory haematophagous organisms that act as vectors of Chagas disease by transmitting the protozoan Trypanosoma cruzi. Their feeding success is strongly related to salivary proteins that allow these insects to access blood by counteracting host haemostatic mechanisms. Proteomic studies were performed on saliva from the Amazonian triatomine bugs: Rhodnius brethesi and R. robustus, species epidemiologically relevant in the transmission of T. cruzi. Initially, salivary proteins were separated by two-dimensional gel electrophoresis (2-DE). The average number of spots of the R. brethesi and R. robustus saliva samples were 129 and 135, respectively. The 2-DE profiles were very similar between the two species. Identification of spots by peptide mass fingerprinting afforded limited efficiency, since very few species-specific salivary protein sequences are available in public sequence databases. Therefore, peptide fragmentation and de novo sequencing using a MALDI-TOF/TOF mass spectrometer were applied for similarity-driven identifications which generated very positive results. The data revealed mainly lipocalin-like proteins which promote blood feeding of these insects. The redundancy of saliva sequence identification suggested multiple isoforms caused by gene duplication followed by gene modification and/or post-translational modifications. In the first experimental assay, these proteins were predominantly phosphorylated, suggesting functional phosphoregulation of the lipocalins.     

The occurrence of species of semi-aquatic Enchytraeidae (Oligochaeta) in Ireland

The Enchytraeidae are essentially terrestrial oligochaetes but many species have marked aquatic tendencies. Over two thirds of recorded Irish species were found in soils which were submerged or frequently flooded and 35% showed a distinct preference for these conditions. Relatively few species were living in soils subject to drought. Red blood was present in 28 species, all but one from soils with more than 55% water. Cognettia sphagnetorum and C. glandulosa developed red blood in very wet conditions. In a survey of Irish wetlands, samples were taken from bog, heath, marsh, fen, margins of lakes and rivers, and saltmarsh. The influence of various environmental parameters was determined using ordination techniques. Magnesium and pH were found to be the most important factors. A high level of magnesium distinguished coastal sites and pH 5.2 separated two clusters representing acid peat and marsh-fen-aquatic sites. Groups of indicator species characterized each of the three clusters. The ecological distribution of the indicator species is described, and their usefulness in classifying enchytraeid communities is discussed.     

Identification of blood species based on surface-enhanced Raman scattering spectroscopy and convolutional neural network

The identification of blood species is of great significance in many aspects such as forensic science, wildlife protection, and customs security and quarantine. Conventional Raman spectroscopy combined with chemometrics is an established method for identification of blood species. However, the Raman spectrum of trace amount of blood could hardly be obtained due to the very small cross-section of Raman scattering. In order to overcome this limitation, surface-enhanced Raman scattering (SERS) was adopted to analyze trace amount of blood. The 785 nm laser was selected as the optimal laser to acquire the SERS spectra, and the blood SERS spectra of 19 species were measured. The convolutional neural network (CNN) was used to distinguish the blood of 19 species including human. The recognition accuracy of the blood species was obtained with 98.79%. Our study provides an effective and reliable method for identification and classification of trace amount of blood.     

Radioimmunoassay of 5-methoxy tryptophol in sheep plasma and pineal glands

No 5-methoxytryptophol (ML) could be detected in sheep plasma using a specific, sensitive radioimmunoassay developed for the purpose. Blood samples were collected from sheep during darkness and daylight and during various stages of the estrous cycle, but in no sample was the ML content above the detection limit of the method. Addition of 1 mM pargyline and neostigmine to blood immediately after collection to block metabolism did not result in a detectable ML concentration. The failure to detect ML was not due to degradation since added ML was not degraded by blood enzymes even after 16 hours incubation at 37 degrees C. Injection of 100 microgram and 1 mg ML sc in sheep resulted in a rapid rise of ML to 95-130pg/ml and 560-1000pg/ml respectively and disappearing with a half life of approximately 15-20 minutes. Sheep pineal glands collected during the light phase contained ML (51 +/- 5pg/mg tissue. X +/- SE, n = 7) which represents less than 6% of the melatonin content. It is concluded that if ML is present in sheep blood it is present at very low levels. It is thus unlikely to be a major circulating pineal hormone in this species, however, its role within the CNS as a local hormone cannot be excluded.     

Properties of Respiratory Pigments in Bimodal Breathing Animals: Air and Water Breathing by Fish and Crustaceans

SYNOPSIS. Oxygen is nearly 30 times more available in air comparedwith water, whereas the carbon dioxide capacity of water is{small tilde}28.5-fold greater than for oxygen, presenting bimodalbreathing species with two very different respiratory milieus.The respiratory pigment plays a variable role in animals switchingbetween the two media. In vertebrates the transition to airbreathing involves two main strategies: a decrease in oxygenaffinity and changes in other haematological parameters suchas haematocrit. When appropriately analyzed, data reveal a decreasein blood oxygen affinity during transition to air. This mayarise via differences in the intrinsic affinity as occurs insome amphibians, or be due to increasesin the organic phosphate:haemoglobin ratio when acclimating to air breathing. Adoptingair breathing often promotes increased haematocrit. It is difficultto discern trends in haemocyanin functioning. Many but not allbimodal and air breathingspecies of crab contain haemocyaninwith high affinity for oxygen. As with haemoglobin there issome tendency for blood haemocyanin concentration to increasewith air breathing but bimodal species are quite variable inthis regard. Different strategies for breathing air are employedby various bimodal crustaceans, some of which involve modulationof haemocyanin oxygenaffinity. The exact mechanisms are oftenspecies dependent and in all bimodal breathing organisms therole of the pigment is best appreciated when the demands ofthe local environment and the behaviour of the species are considered.     

Plasmodium octamerium n. sp., an Avian Malaria Parasite from the Pintail Whydah Bird Vidua macroura*

SYNOPSIS. A new species of avian malaria parasite is described from the pintail whydah Vidua macroura, a very small African finch of the weaver bird family (Ploceidae). Its structure has been studied chiefly in the canary, to which it is easily transmissible by blood inoculation. Since the segmenters most often produce 8 merozoites, the name Plasmodium octamerium n. sp. is proposed. Other characteristics include sexual stages which are usually elongate, often slender, and do not displace the host cell nucleus, and gametocytes indistinguishable from those of many species of Haemoproteus. Erythrocytes are the only blood cells parasitized. The new species resembles Plasmodium fallax in many respects, but gives rise to fewer merozoites and the asexual forms are smaller. Blood-induced infections are also of strikingly different type in some host species. Among susceptible host species are several kinds of finches, pigeons, quail, young chicks, chukars, tree and song sparrows. In most of these hosts infections are mild, but some tree sparrows die as the result of blood infection, and chukars usually die because of massive invasion of the capillary endothelium of the brain by exoerythrocytic forms. These are of the gallinaceum type and may be quite large, producing hundreds of merozoites. Exoerythrocytic stages were sought but not found in other host species.     

What do we measure by a luminol-dependent chemiluminescence of phagocytes?

The review presents a survey of published findings concerning the mechanism of luminol-dependent chemiluminescence in biological systems. The potential of various oxygen species (superoxide anion, hydrogen peroxide, hydroxyl radical) to react with luminol is discussed. The ability of commonly used enzymes (superoxide dismutase, catalase), inhibitors, and oxygen radical scavengers to discriminate between individual oxygen species is assessed together with the potential of a variety of substances encountered in biological systems to interfere in luminol-dependent chemiluminescence reactions. It is concluded that luminol-dependent chemiluminescence gives at present very little ability to discriminate between individual oxygen or radical species. Furthermore, luminol-dependent chemiluminescence used in biological systems is extremely prone to many interferences, which are very difficult to control.     

Estimation of nitric oxide concentration in blood for different rates of generation. Evidence that intravascular nitric oxide levels are too low to exert physiological effects

Endothelium-derived nitric oxide (NO) is a potent vasodilator in the cardiovascular system. Several lines of experimental evidence suggest that NO or NO equivalents may also be generated in the blood. However, blood contains a large amount of hemoglobin (Hb) in red blood cells (RBCs). The RBC-encapsulated Hb can react very quickly with NO, which is only limited by the rate of NO diffusion into the RBCs. It is unclear what the possible NO concentration levels in blood are and how the NO diffusion coefficient (D) and the permeability (Pm) of RBC membrane to NO affect the level of NO concentration. In this study, a steady-state concentration experimental method combined with a spherical diffusion model are presented for determining D and Pm and examining the effect of NO generation rate (V0) and hematocrit (Hct) on NO concentration. It was determined that Pm is 4.5 +/- 1.5 cm/s and D is 3410 +/- 50 microm2/s at 37 degrees C. Simulations based on experimental parameters show that, when the rate of NO formation is as high as 100 nm/s, the maximal NO concentration in blood is below 0.012 nM at Pm = 4.5 cm/s and Hct = 45%. Thus, it is unlikely that NO is directly exported or generated from the RBC as an intravascular signaling molecule, because its concentration would be too low to exert a physiological role. Furthermore, our results suggest that, if RBCs export NO bioactivity, this would be through NO-derived species that can release or form NO rather than NO itself.