Impact of constant storage temperatures and multiple warming cycles on the quality of stored red blood cells.

BACKGROUND: Red blood cells (RBCs) are routinely stored in liquid state at temperatures below 6°C, and RBC unit core temperature should not exceed 10°C during transport. Since the critical temperature of 10°C was chosen mostly arbitrarily, this study investigated the effect of both constant temperature settings as well as multiple rewarming cycles on stored RBCs with respect to morphology, biochemical parameters and haemolysis. MATERIALS AND METHODS: Buffy coat-depleted filtered RBCs were used as standard products. RBCs were stored at 1-6°C (reference group, n = 12), 13 and 22°C (test groups, n = 12 each) or stored at 1-6°C and warmed up five times to 10, 13, or 22°C for a period of 24 h each. Various biochemical parameters were measured weekly. RBCs were further investigated using electron microscopy. RESULTS: Red blood cells stored constantly at 13 or 22°C showed stable haemolysis rates until day 28 and day 14, respectively. RBCs stored at 1-6°C with five warming-up periods to 10, 13 or 22°C each lasting 24 h (total 120 h) did not exceed the limit of the haemolysis rate at the end of storage. Differently shaped erythrocytes were found in all samples, but more crenate erythrocytes appeared after 42 days of storage independent of temperature profiles. CONCLUSION: Red cells can be kept at constant temperatures above 6°C without apparent harmful effects at least until day 14, whereas multiple warming cycles for no longer than 24 h at 10, 13 or 22°C with subsequent cooling do not cause quality loss as assessed using the in vitro assays employed in this study.

Application of elemental microanalysis to elucidate the role of spherites in the digestive gland of the helicid snail Chilostoma lefeburiana.

In this case study we present an application of different analytical electron microscopic methods in biology, to elucidate their usefulness in such investigations. Using analytical electron microscopy, spherites in the digestive gland cells of the helicid snail Chilostoma lefeburiana were examined at three stages: just before the non-feeding period of over-wintering in November, in the middle of over-wintering in February and at its end in March. A detailed characterization of changes in the elemental composition of the spherites was characterized by a combination of transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDXS), electron energy-loss spectroscopy (EELS) and energy filtering TEM (EFTEM). During over-wintering, the spherites passed the following changes. Before over-wintering in November, they consisted of striking concentric layers of electron-dense and electron-lucent zones, while in February and March they showed clear empty zones between materials of different electron density. In November spherites, C, O, Ca, P, Cl, Fe, Si, Na, K, Mg and S were detected, whereas in February ones C, O, N, Cl, Si and S were found and only C, O, N, Si and Cl were detected in March spherites. It is suggested that the elements missing in February and March were used in different physiological processes during over-wintering, like (1) the maintenance of the appropriate elemental composition of the internal environment, (2) accumulation of non-toxic waste materials that cannot be metabolized and (3) avoiding potential intoxication by contamination with toxic metals.

Experimental gastritis in mice enhances anxiety in a gender-related manner.

There is a gender-related comorbidity of pain-related and inflammatory bowel diseases with psychiatric diseases. Since the impact of experimental gastrointestinal inflammation on the emotional-affective behavior is little known, we examined whether experimental gastritis modifies anxiety, stress coping and circulating corticosterone in male and female Him:OF1 mice. Gastritis was induced by adding iodoacetamide (0.1%) to the drinking water for at least 7 days. Inflammation was assessed by gastric histology and myeloperoxidase activity, circulating corticosterone determined by enzyme immunoassay, anxiety-related behavior evaluated with the elevated plus maze and stress-induced hyperthermia tests, and depression-like behavior estimated with the tail suspension test. Iodoacetamide-induced gastritis was associated with gastric mucosal surface damage and an increase in gastric myeloperoxidase activity, this increase being significantly larger in female mice than in male mice. The rectal temperature of male mice treated with iodoacetamide was enhanced, whereas that of female mice was diminished. The circulating levels of corticosterone were reduced by 65% in female mice treated with iodoacetamide but did not significantly change in male mice. On the behavioral level, iodoacetamide treatment caused a decrease in nocturnal home-cage activity, drinking and feeding. While depression-related behavior remained unaltered following induction of gastritis, behavioral indices of anxiety were significantly enhanced in female but not male mice. There was no correlation between the estrous cycle and anxiety as well as circulating corticosterone. Radiotracer experiments revealed that iodoacetamide did not readily enter the brain, the blood-brain ratio being 20:1. Collectively, these data show that iodoacetamide treatment causes gastritis in a gender-related manner, its severity being significantly greater in female than in male mice. The induction of gastritis in female mice is associated with a reduction of circulating corticosterone and an enforcement of behavioral indices of anxiety. Gastric inflammation thus has a distinct gender-dependent influence on emotional-affective behavior and its neuroendocrine control.

Increase in gastric acid-induced afferent input to the brainstem in mice with gastritis.

Acid challenge of the gastric mucosa is signaled to the brainstem. This study examined whether mild gastritis due to dextrane sulfate sodium (DSS) or iodoacetamide (IAA) enhances gastric acid-evoked input to the brainstem and whether this effect is related to gastric myeloperoxidase activity, gastric histology, gastric volume retention or cyclooxygenase stimulation. The stomach of conscious mice was challenged with NaCl (0.15 M) or HCl (0.15 and 0.25 M) administered via gastric gavage. Two hours later, activation of neurons in the nucleus tractus solitarii (NTS) was visualized by c-Fos immunocytochemistry. Gastritis was induced by DSS (molecular weight 8000; 5%) or IAA (0.1%) added to the drinking water for 7 days. Relative to NaCl, intragastric HCl increased the number of c-Fos protein-expressing cells in the NTS. Pretreatment with DSS or IAA for 1 week did not alter the c-Fos response to NaCl but significantly enhanced the response to HCl by 54 and 74%, respectively. Either pretreatment elevated gastric myeloperoxidase activity and induced histological injury of the mucosal surface. In addition, DSS caused dilation of the gastric glands and damage to the parietal cells. HCl-induced gastric volume retention was not altered by IAA but attenuated by DSS pretreatment. Indomethacin (5 mg/kg) failed to significantly alter HCl-evoked expression of c-Fos in the NTS of control, DSS-pretreated and IAA-pretreated mice. We conclude that the gastritis-evoked increase in the gastric acid-evoked c-Fos expression in the NTS is related to disruption of the gastric mucosal barrier, mucosal inflammation, mucosal acid influx and enhanced activation of the afferent stomach-NTS axis.

A contribution to the functional morphology of the midgut gland in phalangiid harvestmen Gyas annulatus and Gyas titanus during their life cycle.

The structure of the midgut gland and its changes in different seasons have been examined in the harvestmen Gyas annulatus and Gyas titanus (Arachnida: Opiliones: Phalangiidae). In both species, in the epithelium of the midgut gland two different types of cells are present: secretory and digestive ones. The secretory cells are characterized by plentiful rER and secretory granula. The digestive cells are characterized by an apical system of tubules. Both cells are connected by prominent specialized junctions. If a secretory cell is in contact with a digestive cell, rER cisterna are in close vicinity and parallel to these junctions. As found light- and electron microscopically and also histochemically, glycogen and lipids are stored in both cells. In both species, glycogen was seen to be used as energy compound during overwintering. At the end of their life, the digestive cells develop into excretory ones, containing metabolic wastes.

A contribution to the functional morphology of the femoral chordotonal organ in the green lacewing Chrysoperla carnea (Neuroptera).

The femoral chordotonal organ (FCO) and the subgenual organ (SGO) of the green lacewing Chrysoperla carnea were examined by conventional light and confocal laser scanning microscopy in order to search for neuroactive substances which are used for neurotransmission in sensory cells of these organs. Antibodies against serotonin, histamine and choline acetyltransferase were tested immunohistochemically. In the FCO, antiserum against serotonin strongly labelled cell bodies and axons of about 16 sensory cells. In the proximal scoloparium all 12 sensory cells showed immunoreaction with antiserotonin. In the distal scoloparium only four of 40 sensory cells were immunoreactive. These results suggest that different neuroactive substances are employed as neurotransmitters in the FCO of the green lacewing and that the proximal scoloparium and the distal scoloparium are functionally differentiated. Contrary to the FCO in the locust, acetylcholine was not found as a neurotransmitter in the FCO of the green lacewing. Additionally, histamine showed a negative result in the sensory cells of the FCO. Other neuroactive substances seem to be used as transmitters in the SGO because none of the tested antibodies showed positive reaction.

Capsaicin-sensitive extrinsic afferents are involved in acid-induced activation of distinct myenteric neurons in the rat stomach.

Challenge of the rat gastric mucosa with 0.5 mol L(-1) HCl activates nitrergic neurons in the myenteric plexus as visualized by c-Fos immunohistochemistry. In the present study, we characterized the activated neurons more extensively by their chemical coding and investigated whether a neural pathway that involves capsaicin-sensitive extrinsic afferents and/or cholinergic neurons transmitting via nicotinic receptors contributes to the activation of myenteric neurons. In multiple labelling experiments, c-Fos was examined for co-localization with nitric oxide synthase (NOS), vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), enkephalin (ENK), gastrin-releasing peptide (GRP), substance P (SP), calbindin D-28k (CALB) and neurofilament 145 (NF 145). All c-Fos-positive neurons were immunoreactive for NOS, VIP, NPY and NF 145, but not for SP, ENK, GRP and CALB. Nerve fibres co-expressing NOS, VIP and NPY were predominantly found in the external muscle layer and in the muscularis mucosae but rarely in the mucosa. Pre-treatment with capsaicin or hexamethonium or a combination of both pre-treatments reduced HCl-induced c-Fos expression by 54, 66 and 63%, respectively. Acid challenge of the stomach, therefore, leads to activation of presumably inhibitory motor neurons responsible for muscle relaxation. Activation of these neurons is partly mediated by capsaicin-sensitive afferents and involves ganglionic transmission via nicotinic receptors.

Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity.

A variety of growth factors promote the complex multistep process of angiogenesis. The mitogenic activity of vascular endothelial growth factors (VEGFs) and placental growth factors (PlGFs), known as cytokines acting predominantly on endothelial cells, was tested on human umbilical vein endothelial cells (HUVEC) and microvascular endothelial cells (MIEC) and compared with the potency of the universally acting basic fibroblast growth factor (FGF-2). The cells were seeded at different cell numbers and incubated with various doses of growth factors for a period of 24-72 h in culture medium +/- serum. Proliferation was determined by measuring the optical density after staining the cells with the tetrazolium salt WST-1. VEGF121 and VEGF165 increased the number of HUVEC and MIEC at low and high seeding densities various doses and incubation times. The efficiency of FGF-2 was less pronounced at high seeding densities of the cells under serum-free conditions. PlGF-1 and PlGF-2 stimulated mitogenesis on HUVEC only at low cell numbers and after a short incubation time by 125 +/- 3% and 102 +/- 5% (P < 0.001), respectively. Longer incubation times with the lower seeding density in the absence of FCS did not induce a significant stimulatory effect of the PlGFs. MIEC responded stronger to all growth factors. In particular under serum free conditions, PlGF-1 and PlGF-2 effectively stimulated cell proliferation by 247 +/- 54% (P < 0.01) and 288 +/- 40% (P < 0.05) at low cell numbers, and by 81 +/- 13% (P < 0.05) and 49 +/- 13% (P < 0.01), respectively, at high cell numbers. The addition of fetal calf serum caused a reduced proliferative response of all growth factors on both cell types related to the controls. In conclusion, MIEC and HUVEC differ in their proliferative response to VEGFs, PlGFs and FGF-2.

Cooperation of NMDA and tachykinin NK(1) and NK(2) receptors in the medullary transmission of vagal afferent input from the acid-threatened rat stomach.

Noxious challenge of the rat gastric mucosa by hydrochloric acid (HCl) is signaled to the nucleus tractus solitarii (NTS) and area postrema (AP). This study examined the participation of glutamate and tachykinins in the medullary transmission process. Activation of neurons was visualized by in situ hybridization autoradiography of c-fos messenger RNA (mRNA) 45 min after intragastric (IG) administration of 0.5 M HCl or saline. IG HCl caused many neurons in the NTS and some neurons in the AP to express c-fos mRNA. The NMDA glutamate receptor antagonist MK-801 (2 mg/kg), the NK(1) tachykinin receptor antagonist GR-205,171 (3 mg/kg) and the NK(2) receptor antagonist SR-144,190 (0.1 mg/kg) failed to significantly reduce the NTS response to IG HCl, whereas the triple combination of MK-801, GR-205,171 and SR-144,190 inhibited it by 45--50%. Only in rats that had been preexposed IG to HCl 48 h before the experiment was MK-801 alone able to depress the NTS response to IG HCl. In contrast, the c-fos mRNA response in the AP was significantly augmented by MK-801, an action that was prevented by coadministration of GR-205,171 plus SR-144,190. Inhibition of neuronal nitric oxide synthase with 7-nitroindazole (45 mg/kg) was without effect on the IG HCl-evoked c-fos mRNA expression in the NTS and AP. Our data show that glutamate acting via NMDA receptors and tachykinins acting via NK(1) and NK(2) receptors cooperate in the vagal afferent input from the acid-threatened stomach to the NTS and participate in the processing of afferent input to the AP in a different and complex manner. These opposing interactions in the AP and NTS and the increase in NMDA receptor function in the NTS after a gastric acid insult are likely to have a bearing on the neuropharmacology of dyspepsia.

Gold toning preserves integrity of silver enhanced immunogold particles during osmium tetroxide treatment for demonstration of a biogenic amine.

We describe a protocol that enhances immunolabelling of nervous tissue for ultrastructural study. Insect tissue is fixed, sectioned, and labelled with a polyclonal antiserum against serotonin and a secondary antibody conjugated with 1 nm colloidal gold. The gold particles are silver-enhanced to ease detection and then protected by gold toning. Finally, the tissue is post fixed in glutaraldehyde fixative followed by osmium tetroxide and further processed for electron microscopy. We demonstrated on insect nervous tissue that gold toning protects marker particles from the influence of osmium tetroxide. Use of buffered solutions throughout the protocol led to well preserved ultrastructural details, and marker particle size was not reduced with a short gold toning time. We also suggest use of this protocol for vertebrate or other invertebrate tissue.

A tungsten supplemented diet attenuates bacterial translocation in chronic portal hypertensive and cholestatic rats: role of xanthine dehydrogenase and xanthine oxidase.

BACKGROUND: Bacterial translocation (BT) plays a major role in the pathophysiological process of spontaneous infections in portal hypertension (PH) and cholestatic jaundice. The major mechanisms promoting BT in experimental animal models are the disruption of the intestinal ecological equilibrium and disruption of the intestinal mucosal barrier. The enzymes xanthine dehydrogenase (XD) and xanthine oxidase (XO) are often implicated as a significant source of oxidants which have a major impact on the impairment of intestinal barrier function. AIM: To investigate the incidence of BT in rats with PH and obstructive jaundice, and to evaluate the impact of XD and XO. METHODS: Animals were subjected to sham laparotomy (SL), PH by calibrated stenosis of the portal vein, and common bile duct ligation (CBDL). They were fed either a standard pellet diet or a tungsten supplemented molybdenum-free diet. Four weeks after the operative procedure, intestinal colonisation and BT to portal vein, vena cava, mesenteric lymph nodes, liver, and spleen were determined. Intestinal XD and XO activity were measured enzymatically and histochemically. RESULTS: Significant (p<0.01) intestinal bacterial overgrowth was present in all PH and CBDL groups compared with the SL group. In normally fed animals after SL, BT occurred in 12%. In PH and after CBDL, the rate of BT increased significantly (p<0.05) to 28% and 54% respectively. In the jejunum of normally fed animals subjected to PH or CBDL, a significant increase in XO was observed (p<0.01). Animals fed a tungsten supplemented diet showed a significant attenuation of BT to 14% in PH and 22% after CBDL (p<0. 05). Tungsten treatment completely suppressed jejunal XD and XO activities. CONCLUSIONS: Significant intestinal bacterial overgrowth, BT, and XD to XO conversion occurred in PH and after CBDL. XD and XO inactivation by a tungsten supplemented molybdenum-free diet significantly reduced the incidence of BT without affecting intestinal bacterial overgrowth. These data strongly support the hypothesis that increased XD to XO conversion may contribute to intestinal barrier failure in PH and after CBDL.

Serotonin-immunoreactive neurones in the visual system of the praying mantis: an immunohistochemical, confocal laser scanning and electron microscopic study.

The distribution, number, and morphology of serotonin-immunoreactive (5-HTi) neurones in the optic lobe of the praying mantis Tenodera sinensis were studied using conventional microscopy and confocal laser scanning microscopy. Five or six 5-HTi neurones connect the lobula complex with the medulla, and at least 50 5-HTi neurones appear to be confined to the medulla. In addition, a few large 5-HTi processes from the protocerebrum supply the lobula complex, and two large 5-HTi processes from the protocerebrum ramify in the medulla and lamina, where they show wide field arborisations. In order to provide a basis for understanding the action of serotonin in the lamina, the ultrastructure of its 5-HTi terminals was examined by conventional and immunohistochemical electron microscopy. The 5-HTi profiles were filled with dense core vesicles and made synapses. Output synapses from 5-HTi profiles outnumbered inputs by about 3 to 1. The terminals of the 5-HTi neurones were in close contact with cells of various types, including large monopolar cells, but close apposition to photoreceptor terminals was rare, and no synapses were found between 5-HTi terminals and photoreceptor terminals.

Femoral chordotonal organ in the legs of an insect,Chrysoperla carnea(Neuroptera).

The femoral chordotonal organ (FCO) inChrysoperla carneais situated in the distal part of the femur and consists of two scoloparia, which are fused at their distal end. The distal scoloparium contains 17-20 scolopidia, and the proximal one six scolopidia. Each scolopidium consists of two sensory cells and three types of enveloping cells (glial, scolopale and attachment cell). The sensory cells of different scolopidia do not lie at the same level in the FCO. Therefore the attachment cells of different scolopidia have different lengths. In the FCO, three types of ciliary roots are found in different sensory cells. The dendrite of the sensory cell terminates in a distal process, which has the structure of a modified cilium (9x2+0). The very distal part of the cilium is surrounded by an extracellular electron dense material, the cap, and ends in a terminal dilation. The scolopale cell contains the electron dense scolopale rods, consisting of plentiful microtubules. In their middle third the scolopale rods are fused and form the scolopale. In the FCO septate junctions, desmosomes and hemidesmosomes are found.

Gastric acid-evoked c-fos messenger RNA expression in rat brainstem is signaled by capsaicin-resistant vagal afferents.

BACKGROUND & AIMS: Gastric acid is known to contribute to ulcer pain, but the mechanisms of gastric chemonociception are poorly understood. This study set out to investigate the pathways and mechanisms by which gastric acid challenge is signaled to the brain. METHODS: Neuronal excitation in the rat brainstem and spinal cord after intragastric administration of HCl (0.35-0.7 mol/L) was examined by in situ hybridization autoradiography for the immediate early gene c-fos. RESULTS: Gastric acid challenge did not induce c-fos transcription in the spinal cord but caused many neurons in the nucleus tractus solitarii and area postrema to express c-fos messenger RNA (mRNA). The HCl concentration-dependent excitation of medullary neurons was in part associated with behavioral manifestations of pain but not directly related to the acid-induced injury and contraction of the stomach. Subdiaphragmatic vagotomy suppressed the c-fos mRNA response to intragastric acid, and morphine inhibited it in a naloxone-reversible manner, whereas pretreatment of rats with capsaicin was without effect. CONCLUSIONS: Gastric acid challenge is signaled to the brainstem, but not the spinal cord, through vagal afferents that are sensitive to acid but resistant to capsaicin. It is hypothesized that the gastric acid-induced c-fos transcription in the brainstem is related to gastric chemonociception.

Characterization of deposits in human lung tissue by a combination of different methods of analytical electron microscopy.

Energy-filtering transmission electron microscopy (EFTEM) was used for imaging of deposits in anthracotic areas of human lung tissue. Unstained ultrathin sections were investigated with a Philips CM20 operated at 200 kV acceleration voltage and equipped with a GATAN imaging filter and an X-ray detector for correlative analysis. The distribution of soot particles in the anthracotic areas could be visualized by recording C-K elemental maps, and inorganic particles between the soot by recording C-K jump ratio images. They could be identified as the mineral muscovite and as an iron oxide phase, which would have been overlooked and obviously their composition would not have been recognized using conventional TEM investigations with stained ultrathin sections. Oxide phases of the inorganic particulates were imaged by recording O-K elemental maps, and silicate and Fe phases with Si-L23 and Fe-L23 jump ratio images, respectively. The interpretation of the elemental maps was supported by recording EEL and EDX spectra from interesting specimen regions. Electron diffraction patterns were used to characterize the mineral crystals.

Mediation by 5_hydroxytryptamine of the femoral vasoconstriction induced by acid challenge of the rat gastric mucosa.

1. Gastric mucosal barrier disruption in the presence of luminal acid causes femoral vasoconstriction via a pathway that appears to be stimulated by messengers generated in the injured gastric mucosa. This study was undertaken to analyse the gastric factors that are responsible for the femoral vasoconstrictor response. 2. Gastric mucosal barrier disruption in the presence of luminal acid was induced by perfusing the stomach of urethane-anaesthetized rats with ethanol (15 %) in 0.01-0.15 M HCl. Blood flow in the left gastric and right femoral artery was estimated by the ultrasonic transit time shift technique. 3. Gastric perfusion of ethanol in HCl caused loss of H+ ions from the gastric lumen, decreased the HCO3- concentration in hepatic portal vein blood, induced macroscopic histological damage to the gastric mucosa, dilated the left gastric artery and constricted the femoral artery. These responses were related to the HCl concentration in the ethanol-containing perfusion medium. 4. The femoral vasoconstriction was also seen when, instead of ethanol, taurocholate (20 mM) was used to disrupt the gastric mucosal barrier in the presence of 0.15 M HCl. 5. The femoral vasoconstriction evoked by gastric perfusion of ethanol in HCl was left unaltered by pharmacological blockade of gastrin and histamine receptors. In contrast, the 5-hydroxytryptamine 5-HT1/2 receptor antagonist methiothepin, but not the 5-HT2A receptor antagonist ketanserin or the 5-HT3 receptor antagonist granisetron, inhibited the ability of both 5-hydroxytryptamine and gastric acid back-diffusion to constrict the femoral artery. 6. Gastric acid back-diffusion caused release of 5-hydroxytryptamine into the gastric lumen, which was related to the HCl concentration in the ethanol-containing perfusion medium. 7. These data show that femoral vasoconstriction evoked by gastric mucosal barrier disruption depends on back-diffusion of acid into the mucosa. The acid-induced damage results in release of 5-hydroxytryptamine from the gastric mucosa, and the pathway leading to constriction of the femoral artery involves 5-hydroxytryptamine acting via 5-HT1/2 receptors as a messenger molecule.

Johnston's organ and central organ in Nezara viridula (L.) (Heteroptera, Pentatomidae).

The fine structure of Johnston's organ and central organ in Nezara viridula (Heteroptera, Pentatomidae) is described. Johnston's organ consists of 45 scolopidia distributed around the periphery of the distal part of the third antennal segment (distal pedicellite). The scolopidia are anchored separately in invaginations of joint cuticle between the pedicel and flagellum. The scolopidia are amphinematic and each scolopidium comprises three sensory cells and three enveloping cells. The latter are a proximal scolopale cell with a typical labyrinth, an attachment cell filled with many microtubules, and a distal accessory cell also filled with microtubules. Axons of 17 scolopidia gather and join one antennal nerve; 28 scolopidia of the opposite side, extend axons into the other antennal nerve. The central organ consists of seven mononematic scolopidia, which comprise of one or two sensory cells. They anchor in the same joint as the scolopidia of Johnston's organ. The sensory cell bodies of the central organ are located close to the antennal nerves, more proximally than those of Johnston's organ. The axons of four scolopidia join one antennal nerve and those of the remaining three scolopidia join the other antennal nerve. Enveloping cells similar to those in Johnston's organ are present in the central organ.

Morphologic basis of the functional gastric acid barrier.

The gastric mucosa possesses a functional barrier that prevents intrusion of luminal acid. The aim of the present study was to elucidate the morphologic basis of this barrier by exploring the effect of acid challenge on the gastric mucosal epithelium, basal lamina, and microvasculature. The stomachs of urethane-anesthetized rats were perfused, for at least 45 minutes, with 0.05 M HCl or 0.15 M HCl in the absence or presence of the mucosal barrier breaker ethanol (15%) and examined by light, scanning, and transmission electron microscopy. Gastric perfusion with 0.05 M HCl alone caused superficial cell injury, the damaged surface cells loosing the surface membrane, whereas the junctional complex and basolateral membrane were preserved. Perfusion of 0.15 M HCl alone led to focal ablation of surface epithelial cells in the interfoveolar regions, and cells in the gastric pits remained grossly normal. Exposure to the barrier breaker ethanol (15%) in the presence of 0.05 M HCl caused extensive ablation of the surface epithelium. There were many focal areas in which the honeycomb structure of the lamina propria was exposed and the basal lamina was removed. Gastric mucosal damage progressed further when the luminal HCl concentration was raised to 0.15 M in the presence of ethanol (15%). In this instance, extensive areas with deep erosions and vast areas of deep-reaching ablation of the epithelium and basal lamina were observed. Ultrastructurally, there was extensive damage to the endothelium of capillaries lying underneath denuded areas of the gastric mucosa, the injured capillaries containing erythrocyte ghosts and thrombocyte aggregates. The data suggest that the integrity of the junctional complex, basolateral membrane, and basal lamina forms the morphologic basis of the functional gastric acid barrier. Once these structures are disrupted or ablated to an appreciable extent, damage forms in the mucosal microvasculature, and injury progresses to deeper layers of the mucosa.

Structure of the subgenual organ in the green lacewing, Chrysoperla carnea.

REM and TEM studies of the subgenual organ in Chrysoperla carnea (Neuroptera: Chrysopidae) show that it is composed of three scolopidia, each with one sensory, one scolopale and one cap cell. The distal part of the dendrite shows a cilium with a '9 + 0' structure. The cross-handing pattern of the ciliary root has a periodicity of bands of about 61 nm. The scolopale material in a certain part of the scolopale cell is organized into five rods. The cell bodies of all three cap cells form a lens-like structure. the velum, which is fixed to the leg wall and the trachea with an extracellular material. The importance of the velum is discussed. Four types of intercellular junction are found; spot desmosomes. belt desmosomes, septate junctions and gap junctions.

Ultrastructural localization of wheat germ agglutinin binding sites on the sperm surface of water buffalo (Bubalus bubalis). A fracture label study.

In the present study we have examined the plasma membrane surface organization employing fluorescein isothiocyanate linked wheat germ agglutinin (WGA) of the cauda epididymal and ejaculated spermatozoa of water buffalo. Intramembrane particle distribution pattern in the various segments of the spermatozoa has also been observed. WGA-ovomucoid gold has been used to study the distribution of sialoproteins on the sperm surface. With fracture label, WGA receptor sites have been identified on the fractured membrane halves of the sperm plasma membrane overlying the acrosome as well as the middle piece and the principle piece.