Polarization optical analysis of blood cell membranes

The present study deals with investigations of membrane structure using polarization topo-optical reactions. Polarization microscopy is a special field of biological submicroscopic morphology. It represents a powerful tool well able to reveal the features of organization of biological structures, and the regularity of macromolecules building cells and tissues - properties that cannot directly be studied by other approaches to complex biological systems. Only in "pure" systems can X-ray diffraction, or the analysis of circular dichroism and the dispersion of optical rotability provide data equivalent to those obtained by polarization microscopy in complex systems. One of the main drawbacks of molecular biology is that most information is relevant to isolated, purified particles or macromolecules. Thus, no conclusions can be drawn concerning the original arrangement of molecules. The gap between biochemical-biophysical and morphological approaches to molecular arrangement in complex structures is bridged by the polarization optical technique. As was pointed out in the introduction, polarization microscopy became a routine biological research method following the pioneering work of Romhányi. His enlightening topo-optical reactions (Romhányi 1960, 1963, 1966) were based on the oriented dye binding of the original charge carriers of regularly arranged tissue constituents. The second group of Romhányi's topo-optical reactions comprised procedures such as sulfation (Romhányi et al. 1973, 1974), the aldehyde-bisulfite-toluidine blue (ABT) reaction (Romhányi et al. 1974, 1975), the permanganate-bisulfite-toluidine blue (PBT) reaction (Fischer 1979, 1979a), and the sialic acid-specific reaction (Makovitzky 1980) all of which operate with induced dye-binding groups; i.e. dye-binding moieties on biological macromolecules are produced by specific chemical reactions.     

喀麦隆塞玛岭地区溪流中的水生丝孢菌区系

Aquatic hyphomycetes on submerged fallen leaves and deadwoods have been numerously reported in fast running streams in temperate countries(Ingold,1976;Ingold,1979;Chauvet,1990;Barlocher & Rosset,1987;Barlocher et al.,1995;Descals et al.,1995).However,documented information is considerably limited in African countries(Ingold,1956;Dixon,1959;Le-John,1965;Ferreira et al.,1981),and unavailable in Cameroon,a country mostly covered with heavy tropical forests(Loung,1980).This paper is to present a list of aquatic and aeroaquatic hyphomycetes identified from foam samples collected in Cameroon during a two-year survey.     

痛风的临床病理特征分析并文献复习

目的:总结痛风临床病理特点。方法:回顾性分析1例痛风患者的生化机制、临床病理特征、刚果红染色、PAS染色特点、鉴别诊断要点,并复习相关文献。结果:患者主要临床表现为间断性多关节肿痛3年,加重伴发热3个月。体格检查发现患者有多发性皮下结节、多关节肿胀压痛。左腕、左肘皮下结节活检,经HE染色后光镜查见大量肉芽肿性病变,有的多核巨细胞内查见被吞噬的异物,有的病灶尚查见呈均质状物(尿酸盐结晶),其周围有较多异物巨细胞及纤维结缔组织包绕呈结节状,在结节的周边纤维血管周围可查见残留分化成熟的淋巴细胞及少数嗜酸性粒细胞。刚果红、PAS染色均为阴性。偏光显微镜下,刚果红未查见绿色强折光晶体,但见多量略呈淡黄色具有强折光性的晶体呈棒状或梭形。结论:痛风在刚果红染色偏光显微镜下观察呈淡黄色梭形或针状结晶,具有强折光性晶体,但这是否是痛风在刚果红染色的特征尚有待于进一步研究。     

Histochemistry and functional significance of putative neocortical transmitter substances: a review

1. Intrinsic neuronal chains of the neocortex communicate most probably with amino acid transmitters. These involve both excitatory (glutamate, aspartate--Nadler et al. 1976) both inhibitory (GABA--Ribak 1978) amino acids, and ensure fast, ionotropic postsynaptic actions (Eccles, McGeer 1979). 2. Some interneurons of the neocortex seemingly operate with the peptide transmitter VIP (Lorén et al. 1979). Presumably, this is a metabotropic, slowly acting substance (Dodd, Kelly and Said 1979). 3. The existence of intrinsic cholinergic neurons in the neocortex is a matter of question (Krnjevic and Silver 1965). It is worth to mention that in the periphery, cholinergic terminals also contain and release VIP (H?kfelt et al. 1980). It is not known, whether this transmitter dualism can be found in neocortex, too. An ascending cholinergic system projecting from the basal forebrain to the neocortex exists and exerts profound influence on cortical function (Shute and Lewis 1967). 4. Diffusely terminating, ascending monoamine axons innervate the neocortex and modulate interneuronal transmission (Thiery et al. 1977; Morrison et al. 1981, Lidov et al. 1981). 5. The neuropeptide SP excites cortical neurons (Phillis and Limacher 1974), and its presence in thin axons can be demonstrated immunohistochemically (H?kfelt et al. 1976). 6. Neocortical efferents to the thalamus and striatum seemingly use glutamate or aspartate (Fonnum et al. 1981). The transmitters of other corticofugal projections are not known. 7. The transmitters of specific thalamic afferents and those of callosal and association projections are unknown, too. 8. The main task of future histochemistry is to explore the synaptology of neocortical neurons and afferent systems with identified or evidenced transmitters, viz. to explore the neurochemical subsystems of cortical organization. The tool for it could be the immunohistochemistry, and future development depends mainly on the synthesis and purification of suitable antigens. The knowledge on the synaptology of identified neurochemical units of the cortex would be the basis of the understanding at least partly of the pharmacological effects exerted by the putative neocortical transmitters.     

Do enzyme activities vary along muscle fibres?

Summary Distribution of succinate dehydrogenase activity along muscle fibres has been studied qualitatively by histochemistry on single microdissected rat muscle fibres and quantitatively by comparative kinetic microphotometry on longitudinal muscle sections. Qualitative staining reactions showed no appreciable variations in enzyme activity along the fibres regardless of fibre type. By quantitative assessment, minor variations were found along fibres but were within the range of the experimental error. These variations are of the same magnitudes as those observed in enzyme activities of pieces of the same fibre by means of quantitative microchemical methods performed in our laboratory (Spamer and Pette 1979; Nemeth et al. 1980a, b). Our results provide evidence that the enzyme levels are the same along the course of a muscle fibre.     

Amyloid from a histochemical perspective. A review of the structure,properties and types of amyloid,and a proposed staining mechanism for Congo red staining

Amyloid is a diverse group of unrelated peptides or proteins that have positive functionality or are associated with various pathologies. Despite vast differences, all amyloids share several features that together uniquely define the group. 1) All amyloids possess a characteristic cross-ß pattern with X-ray diffraction typical of ß-sheet secondary protein structures. 2) All amyloids are birefringent and dichroic under polarizing microscopy after staining with Congo red, which indicates a crystalline-like (ordered) structure. 3) All amyloids cause a spectral shift in the peak wavelength of Congo red with conventional light microscopy due to perturbation of π electrons of the dye. 4) All amyloids show heightened intensity of fluorescence with Congo red, which suggests an unusual degree of packing of the dye onto the substrate. The ß portion of amyloid molecules, the only logical substrate for specific Congo red staining under histochemical conditions, consists of a stack of ß-sheets laminated by hydrophilic and hydrophobic interactions between adjacent pairs. Only the first and last ß-sheets are accessible to dyes. Each sheet is composed of numerous identical peptides running across the width of the sheet and arranged in parallel with side chains in register over the length of the fibril. Two sets of grooves are bordered by side chains. X grooves run perpendicular to the long axis of the fibril; these grooves are short (the width of the sheet) and number in the hundreds or thousands. Y grooves are parallel with the long axis. Each groove runs the entire length of the fibril, but there are very few of them. While Congo red is capable of ionic bonding with proteins via two sulfonic acid groups, physical constraints on the staining solution preclude ionic interactions. Hydrogen bonding between dye amine groups and peptide carbonyls is the most likely primary bonding mechanism, because all ß-sheets possess backbone carbonyls. Various amino acid residues may form secondary bonds to the dye via any of three van der Waals forces. It is possible that Congo red binds within the Y grooves, but that would not produce the characteristic staining features that are the diagnostic hallmarks of amyloid. Binding in the X grooves would produce a tightly packed series of dye molecules over the entire length of the fibril. This would account for the signature staining of amyloid by Congo red: dichroic birefringence, enhanced intensity of fluorescence and a shift in visible absorption wavelength.     

Histological, histochemical and ultrastructural studies on the bulbus arteriosus of the sticklebacks, Gasterosteus aculeatus and Pungitius pungitius (Pisces: Teleostei)

The bulbar wall has three layers. Its lining consists of squamous-columnar endothelial cells that store neutral mucopolysaccharides and are PAS-positive. They do not contain large amounts of acid phosphatase, acid mucopolysaccharides, glycogen or lipids. A morphometric analysis shows that 32% of the cell volume in Pungitius and 12% in Gasterosteus is occupied by specific granules, 100–600 nm in diameter. According to X-ray probe micro-analysis, these granules bind chromium ions, even though the endothelial cells do not contain catecholamines. Rootlets, packed with plasmalemmal vesicles, extend from the endothelial cells into the middle layer of the bulbus. Here, smooth muscle cells alternate with elastic fibres. The staining reactions of bulbar elastica are compared with those in the mammalian aorta and the ligamentum nuchae. The outer layer of the bulbus is visceral pericardium and beneath its covering mesothelial cells are numerous collagen fibres, non-myelinated nerves, occasional fibroblasts and melanocytes. Scanning electron microscopy shows that the bulbar lining is thrown into longitudinal folds, but that there are no trabeculae subdividing the lumen.
Many features of the bulbus arteriosus may be related to the low systolic pressures of teleosts and to the proximity of their heart and gills. In contrast to mammals, only a small part of the arterial system can act as a windkessel. The bulbus is thus more distensible than the mammalian aorta and must lie within the pericardial cavity so that its greater excursions can be accommodated. Perhaps because the bulbus is so distensible, it has elastic fibres rather than lamellae. This in turn may affect the organization of the smooth muscle cells which do not form "span muscles" as in some mammalian aortae. Like most cells in the bulbus, they are joined to others by desmosomes. Evidently, firm cohesion is important in highly distensible vessels.     

Onchocerciasis in British cattle: a study of Onchocerca gutturosa and O. lienalis in North Wales

Onchocerca gutturosa and O. lienalis infections in British cattle were studied by examination of cattle post-mortem originating from North Wales and Cheshire (north west England). In 463 adult animals, the microfilarial (mf) prevalence was 28.5%. In 95.3% of the mf infected animals, gravid worms could not be found at either the ligamentum nuchae or the gastro-splenic omentum. Dermal mf at the head were identified as O. gutturosa on the basis of their highly significant association with the presence of gravid O. gutturosa at the ligamentum nuchae, which were found in only 3.2% of cattle. Mfs were isolated from different skin sites and from adult worms and a minimum of 10 mfs from each isolate were examined for width and acid phosphatase (AP) staining pattern. The width of O. gutturosa dermal mf was less than 4 micron (4 isolations), narrower than that of putative O. lienalis mf isolated from umbilical skin of cattle without evidence of O. gutturosa, which were in 20/22 isolations greater than 4 micron wide. The dermal mf were also distinguished on the basis of different AP staining patterns which, for each species, correlated closely with that of hatched intrauterine mf from their respective adult female worms. Based on the criteria of morphology and AP staining patterns the mf species prevalences in the survey population were estimated as O. lienalis 24.1% and O. gutturosa 2.2%, with a further 2.2% of cattle infected with both species. The results indicate that the predilection site of adult O. lienalis is not the gastro-splenic omentum. In North Wales, the distribution of the two species was different; O. lienalis was widely distributed in all cattle rearing areas both lowland and upland, whereas O. gutturosa was largely restricted to valleys close to major rivers.     

STRUCTURAL BIOLOGY OF THE FIBRES OF THE COLLAGENOUS AND ELASTIC SYSTEMS

The different types of fibres of the collagenous and elastic systems can be demonstrated specifically in tissue sections by comparing the typical ultrastructural picture of each of the fibre types with studies using selective staining techniques for light microscopy. A practicalmodus operandi, which includes the recommended staining procedures and interpretation of the results, is presented. Micrographs and tables are provided to summarize the differential procedures. Reticulin fibres display a distinct argyrophilia when studied by means of silver impregnation techniques, and show up as a thin meshwork of weakly birefringent, greenish fibres when examined with the aid of the Picrosirius-polarization method. In addition, electron-microscopic studies showed that reticulin fibres are composed of a small number of thin collagen fibrils, contrasting with the very many thicker fibrils that could be localized ultrastructurally to the sites where non-argyrophilic, coarse collagen fibres had been characterized by the histochemical methods used. The three different fibre types of the elastic system belong to a continuous series: oxytalan—elaunin—elastic (all of the fibre types comprising collections of microfibrils with, in the given sequence, increasing amounts of elastin). The three distinct types of elastic system fibres have different staining characteristics and ultrastructural patterns. Ultrastructurally, a characteristic elastic fibre consists of two morphologically different components: a centrally located solid cylinder of amorphous and homogeneous elastin surrounded by tubular microfibrils. An oxytalan fibre is composed of a bundle of microfibrils, identical to the elastic fibre microfibrils, without amorphous material. In elaunin fibres, dispersed amorphous material (elastin) is intermingled among the microfibrils.     

The alpha 1 chain of type VIII collagen is associated with many but not all microfibrils of elastic fiber system.

The antigen of monoclonal antibodies which had labeled the hexagonal lattice of Descemet's membrane in a specific manner was shown to be the alpha 1 chain of type VIII collagen by immunoblotting followed by amino acid sequence analysis. With this antibody, the localization of alpha 1 (VIII) in various tissues was studied by several immunocytochemical methods. Under light microscopy, the alpha 1 (VIII) was found in a fine fibrillar form in various capsular tissues such as capsules of the liver, kidneys, adrenals, lungs and so on. It was also present in dense connective tissues such as the Achilles tendon, and periodontal and perivertebral ligaments. When some dense connective tissues which had been negative to the label including the intima of aorta, perimysium and Glisson's sheath of the liver, were subjected to pepsin digestion, epitopes were revealed which showed a specific immunofluorescence pattern. In many locations the pattern of localization coincided with that of elastic fiber components, and full or partial colocalization with tropoelastin or costaining with resorcin-fuchsin staining was observed. In immunoelectron microscopy, the antigen (alpha 1 (VIII)) was localized on the surface of, but not inside, elastic fibers. However, some tissues which are rich in elastic fibers or microfibrils remained unlabeled. These included elastic fibers of the aortic media and ligamentum nuchae as well as ciliary zonules. Therefore it is suggested that alpha 1 (VIII) is a collagen associated with microfibrils of some elastic fiber systems, but is not an intrinsic component of either elastic fibers or of microfibrils.     

An alternative model for the structural mechanics of hagfish slime threads

Hagfish slime threads are an interesting intermediate filament (IF) system in that, in contrast to hair, they appear to be a series system of IFs, consisting of rods and terminal domains (TDs), without matrix. The protein composition also differs from hair. Published data show that the wet stress-strain curve consists of four regions: I, increasing low modulus; II, plateau; III increasing higher modulus; IV decreasing modulus to break. Beyond 34% extension, there is plastic deformation, which contrasts with the elastic deformation of most biological fibres. The paper considers two explanations, one published by Fudge et al. [D.S. Fudge, K.H. Gardner, V.T. Forsyth, C. Riekel, J.M. Gosline, Biophys. J. 85 (2003) 2015] and one new, of deformation in the four regions. The former regards the TDs as elastic, entropic coils, which extend in region I. The latter regards the TDs as having a plastic, energy-dependent extension in regions III and IV, which is comparable to the drawing of polyester fibres. A rough theoretical model of jumps over energy barriers gives a similar prediction. Twist is suggested as a mechanism for overall thread cohesion. Experimental and theoretical ways forward to a greater understanding of the structural mechanics of hagfish threads are suggested. The behaviour of the total thread/mucus/water system is discussed and some speculations on the defensive mechanisms that have evolved are presented.     

Anisotropic cross-bands in peritrophic membranes of Diptera

In agreement with previous findings in adult Calliphora erythrocephala a periodic incorporation of glucose-¹⁴C and methionine-S35 has been observed into the peritrophic membranes of C. erythrocephala and Lucilia sericata larvae as well as in peritrophic membranes of adult L. sericata and Musca domestica. The width of the periodic units (=labelled band + unlabelled interval) is of the order of 50 to 80 μm.After staining with the dichroitic dye Congo red, peritrophic membranes exhibit a periodic pattern of anisotropic bands in polarized light, resembling the pattern in autoradiographs. If stained membranes are rotated between crossed polars, every 90° the extinction is shifted from a ‘band’ into an ‘interval’ and vice versa, indicating that in these peritrophic membranes two anisotropic sections are arranged perpendicular to each other in series. There are no differences between band patterns of males and females.A relation between width of the bands and the diameter of the peritrophic membranes seems to be possible, but since correlation factors are only of the order of 0·83 it is not significant. Besides the macrounits (band+interval) with a width of about 50 to 80 μm, smaller units with a width of 10 to 14 μm have been detected only in the peritrophic membranes of adult C. erythrocephala and L. sericata, as well as in the inner membranes of adult M. domestica. The anisotropic structure of these microbands is arranged about 45° with respect to that in macrobands.Peritrophic membranes of a blackfly larva, Odagmia ornata, after staining with Congo red show in polarized light a complicated zig-zag pattern of periodic units with a width of 15 to 20 μm, or patches with a regular orthogonal network.     

The Effects of Praziquantel on the Monogenean Gill Parasite Pseudodactylogyrus Bini

The anthelminthic Praziquantel is found to be effective against trematodes and cestodes (for a comprehensive review of various aspects of this drug, see Andrews et al. 1983). Praziquantel has been reported to affect fish cestodes (Pool et al. 1984, Moser et al. 1986, Ward et al. 1986), fish digeneans (Bylund & Sumari 1981) and fish monogeneans (Schmahl & Mehlhorn 1985). Therefore Praziquantel could be a potential drug against Pseudodactylogyrus spp. parasitizing the gills of eels. Infections with these monogeneans cause problems in eel farms (Ogawa & Egusa 1976, Egusa 1979, Chan & Wu 1984, Mellergaard & Dalsgaard 1986). Köhler & Bachmann (1978) tested the effect of Praziquantel on the activity of succinate dehydrogenase and NADH-diaphorase (NADH-oxidase) from Ascaris suum and reported that the latter enzyme was inhibited by Praziquantel.     

Activation of lymphocytes alters Fc receptor-beta 2-microglobulin interrelationship on the lymphocyte surface

The effect of anti-beta 2-microglobulin (anti-B2Mi) on the expression of Fc receptors (FcR) of human lymphocytes was compared on resting and activated cells. Previously we reported that anti-B2Mi induces a "co-shedding" of FcR with the beta 2-microglobulin (B2Mi)-anti-B2Mi complexes when used under the conditions where the redistribution of membrane molecules is allowed (Sármay et al., Cell. Immunol. 56, 452, 1980; Sármay et al. Immunology 36, 339, 1979). Furthermore our group also described two types of FcR-bearing cells, one which shed their FcR during a temperature shift from 4 to 37 degrees C (FcRI+ cells) and the other which has an immobile type FcR under the same circumstances (FcRI+ cells) (Sándor et al., Immunology 38, 553, 1979; Sármay et al., Immunology 34, 315, 1978). In this work we have characterized the FcR released from the membrane as a consequence of anti-B2Mi treatment. We have found that they are the mobile, FcRI type. It was proved that the shedding of this FcRI is a consequence of the anti-B2MI-induced transformation of FcRII into the FcRI form on the membrane of the antibody-treated lymphocytes. On the activated T cells, however, anti-B2Mi is incapable of inducing the same phenomenon in the early phase of activation. In contrast, FcR expression is blocked by anti-B2Mi treatment similarly to that on resting lymphocytes, on the surface of activated B cells, or on activated T cells in the later phases of activation.     

Characterisation of the isoenzymes of phosphoglucomutase (PGM) determined by the first (PGM1) and second (PGM2) locus observed by isoelectric focusing

Summary The existence of four alleles of phosphoglucomutase (PGM₁) in human red cell lysates has previously been demonstrated by isoelectric focusing (Bark et al., 1976; Kühnl et al., 1977; Sutton and Burgess, 1978). Experiments are now described in which the position of each of the first-locus (PGM₁) and second-locus (PGM₂) isoenzymes is defined, thus extending and confirming the original proposal made by Bark et al.     

Proteomics with Mass Spectrometry Imaging: Beyond Amyloid Typing

Detection and typing of amyloid deposits in tissues are two crucial steps in the management of systemic amyloidoses. The presence of amyloid deposits is routinely evaluated through Congo red staining, whereas proteomics is now a mainstay in the identification of the deposited proteins. In article number 1700236, Winter et al. [Proteomics 2017, 17, Issue 22] describe a novel method based on MALDI–MS imaging coupled to ion mobility separation and peptide filtering, to detect the presence of amyloid in histology samples and to identify its composition, while preserving the spatial distribution of proteins in tissues.     

Giemsa banding patterns in chronic lymphocytic leukaemia.

The banding patterns of chromosomes from 20 patients with chronic lymphocytic leukaemia (C.L.L.) have been analyzed. 97 of 100 metaphases examined had a normal banding pattern. The 3 remaining metaphases, all from one patient had bands similar to those seen after aging. It is concluded that the chromosomes in C.L.L. have normal banding patterns. The majority of cytogenetic studies in chronic lymphocytic leukaemia have reported normal chromosomes (Fitzgerald and Adams 1965; Oppenheim et al., 1965; Lawler et al., 1968). An inherited abnormality of G group chromosome (No. 22) has been reported in a family, three members of whom developed C.L.L. (Fitzgerald and Hamer, 1969), but further investigations of cases of familial leukaemia failed to reveal a similar abnormality (Fitzgerald et. al., 1966). The development of new techniques which allow the positive identification of individual chromosomes (Caspersson et al., 1969; Dutrillaux and Lejeune, 1971; Sumner et al., 1971; Seabright, 1971), has revolutionised human cutogenetics and revealed additional information regarding chromosome abnormalities and leukaemia (Rowley, 1973; Lobb et al., 1972; Milligan and Garson, 1974). The purpose of this investigation was to determine whether the chromosomes in C.L.L. have normal banding patterns.     

The innervation of the hepatic portal vein in the rabbit: Ultrastructural evidence against “purinergic” neurotransmission

Summary The relative density of adrenergic and non-adrenergic nerves in the hepatic portal vein of the rabbit has been determined ultrastructurally. Adrenergic nerves were visualised with the modified chromaffin procedure of Tranzer and Richards (1976). Nearly equal numbers of adrenergic and non-adrenergic nerve profiles were found, indicating a much greater density of innervation by non-adrenergic nerves than that described by Burnstock et al. (1979) using light microscopic histochemical methods. These results imply that part of the argument used by Burnstock et al. (1979) to support purinergic transmission in rabbit portal vein is probably invalid.     

Some observations on the collagen fibrils of the egg capsule of the dogfish, Scyliorhinus canicula

The egg capsule of the dogfish Scyliorhinus canicula is a collagenous material with a laminated, plywood (orthogonal) construction. The collagen fibrils which constitute the bulk of the egg capsule wall have a unique, highly ordered structure (Knight and Hunt, 1974; 1976, 1986; Gathercole et al., 1993) which is thought to represent a smectic A liquid crystalline phase (Knight et al., 1993). The egg capsule is extremely strong and chemically inert (Hunt, 1985). It is stored, secreted and formed by the nidamental gland (Rusaou?n 1976, 1990 a, b; Knight and Feng, 1992). During intracellular storage, secretion and fibrillogenesis, the dogfish egg capsule collagen appears to pass through a remarkable series of textures within a lyotropic liquid crystalline phase diagram (Knight et al., 1993). In the present communication, further observations on the ultrastructure of the collagen fibrils and their arrangement within the laminae of the fully-formed egg capsule are reported. The effect of tilting ultrathin sections of fibrils in the goniometer stage of a transmission electron microscope are described, demonstrating that the crystalline lattice within the fibril appeared twisted more or less regularly into a long pitch helix. Other observations indicated that some of the fibrils were in turn twisted round one another to form fibres which therefore had a coiled-coil structure. The fibres are arranged parallel to one another in the laminae which are stacked to give an orthogonal plywood construction. The effects of staining fibrils with cuprolinic blue and with tannic acid are reported. Reduction in the water content of the fibrils before fixation appeared to move some of the fibrils through the part of the lyotropic phase transition diagram converting them from smectic A to smectic C. Finally, evidence is presented that the fibrils shrank, but remarkably, still retained a longitudinally-ordered but modified, molecular arrangement even after boiling in water for periods of up to 10 min. These observations are discussed in relation to other collagens.