L.D.H. and L.D.H. isoenzymes of the intra-uterine secretions of the cow during the estrous cycle

Uterine secretions were collected from 20 mature cows during estrus (day 0), metestrus (day 5), diestrus (day 10) and proestrus (day-1). Lactate dehydrogenase (LDH) and LDH isoenzymes activity were evaluated. No significant cyclic variations of LDH activity was found in the uterine secretions while the mean of the enzyme activity was higher during the estrogenic period of the cycle. The relative activity of LDH-1, LDH-2 and LDH-3 isoenzymes were higher during proestrus and estrus whereas LDH-5 activity was more important during metestrus. The LDH-3 seems to have the higher relative activity in uterine secretions of the cow.     

An e.p.r. study of the non-equivalence of the copper sites of caeruloplasmin.

The two Type 1 (blue) copper-binding sites of caeruloplasmin were spectroscopically differentiated by the kinetic analysis of the e.p.r. spectra during the redox cycle. One blue copper, with a hyperfine splitting constant (A parallel) of 6.8 mT, which was rapidly reduced, was not reoxidized by oxygen, whereas it was reoxidized by H2O2. The other blue copper (A parallel = 5.8 mT), which was reduced slowly, was rapidly reoxidized by either oxygen or H2O2. A conformational change of the Type 2 copper was concomitant with the fast reduction of Type 1 copper, whereas its reduction occurred during the slow phase. This sequence of events was reversed in the reoxidation step, that is, the Type 2 copper reappeared rapidly as the species with altered conformation and reverted to the symmetry typical of the native state in the slow phase. The specific reaction of a blue-copper site with the H2O2 can tentatively be related to the established ability of caeruloplasmin to prevent 'oxidative' attack of proteins and lipids.     

Pleuromeia from the lower triassic of the far east of the U.S.S.R.

Pleuromeia obrutschewii Elias from Russian Island (Russkiy Ostrov, near Vladivostok) is hardly distinguishable from the European P. sternbergii (Muenster) Corda. The sporangia are adaxial, filling spoon-like depressions of the megasporophylls which have sterile tips. P. olenekensis sp. nov. from the Olenek River (northeastern Siberia) has larger sporangia and much larger megaspores with three-layered walls. The outer layer (ectexosporium) is reticulate. It is assumed that in other species this layer is lacking due to imperfect preservation. Mature megasporophylls, when shed, have a buoy-like shape and are often deposited together with cephalopod shells. This suggests a special mechanism of propagation by means of megasporophylls dispersed by water currents. The cosmopolitan distribution of Pleuromeia points to weakened climatic zonation in the Early Triassic.     

Studies on the maturation of immune responsiveness in the mouse. II. Role of the spleen.

Experiments were designed to investigate the role of the spleen in the development of the murine immune system. By using mice splenectomized within 24 hr of birth, as well as mice with a hereditary, congenital absence of the spleen, the primary immune response to sheep erythrocytes was examined. The immunocompetence of lymph node cells from spleenless or control mice was assessed in vitro, in organ and in cell suspension cultures, and in vivo, by transfer into lethally irradiated syngeneic recipients followed by antigenic stimulation. The immunologic capacities of thymus and bone marrow cells were similarly tested by injection separately or in combination into irradiated syngeneic mice. Lymph node cells from spleenless animals appeared fully competent both in vitro and in transfer experiments. Neither neonatal splenectomy nor congenital absence of the spleen significantly reduced the capacity of bone marrow or thymus cells to participate in the immune response to sheep erythrocytes.     

Vegetative innervation of the esophagus. III. Intraepithelial endings.

Intraepithelial fibers do occur in the mucosa of the esophagus, as demonstrated by the osmium tetroxide-zinc iodide method in cats and rhesus monkeys. The esophagus is divided into three parts, in order to study the penetration incidence, and the uppermost and the lowest show the greatest density of penetration, while in the middle portion only occasional fibers in small numbers are found. The specific characteristics observed in this type of fiber, such as their distribution along the wall of the esophagus, the levels reached by their endings within the mucous epithelium itself and this same epithelium considered as the specified destination of the endings, lead to the belief that they may be functionally regarded as structures of a sensory character.     

Molecular analyses of the Salmonella g. . . flagellar antigen complex.

Salmonella flagellar filaments are polymers of a highly antigenic protein, termed flagellin. Eight main subfactors have been identified in the Salmonella phase-1 g. . . series flagellar antigen. To determine the molecular basis for expression of the epitopes by which the g. . . family subfactors are distinguished, 10 members of this series were selected and their fliC (the structural gene for phase-1 flagellin) genes were sequenced. Comparative analyses of the inferred primary structures of these flagellins did not allow the identification of linear epitopes responsible for the antigen subfactors. This suggests that conformational aspects are involved in determining the antigenic specificity in these cases. A phylogenetic analysis of the flagellin sequences showed that members of the g. . . series do not form a single coherent unit.     

On the nature of P.P.L.O.

Summary In a number of strains of the three species of P.P.L.O. of human origin coccoid elements have been observed, which are Gram positive or Gram variable and of very small size (<0.5 μ). They are able to grow in pure culture on the surface of nutrient agar in very small colonies. Although they have bacterial characteristics, they could not be identified as known bacteria. Agglutination tests are in favour of a serological relationship between the coccoid elements and P.P.L.O. The suggestion that these elements might be an outside contamination is made unlikely by several observations. The supposition that P.P.L.O.s are L forms of these coccoid elements should be considered further.     

Branchiobdellida (Oligochaeta) from the farthest South-East of the U.S.S.R.

Eight species of the genera Branchiobdella and Cirrodrilus are reported from the Soviet Far East. Six of them from the Komarovka Stream in the Ussuriiski State Nature Reserve: Branchiobdella cheni (Liu, 1964), B. domina sp.n., Cirrodrilus chosen (Yamaguchi, 1934), C. suzukii (Yamaguchi, 1934), C. pugnax sp.n., and C. fimbriatus sp.n. Branchiobdellidans are abundant in this stream, occurring on the crayfishes Cambaroides dauricus wladiwostokensis as well as in benthic samples. Two from the northern Amur River basin near Khabarovsk: Branchiobdella minuta Pierantoni, 1912, and Cirrodrilus quadritentacularis (Liu, 1984), living on the crayfishes Cambaroides dauricus dauricus and C. schrencki . The genus Cirrodrilus demonstrates several trends of parallel variation on the Asian continent and the Japanese Islands; the more apomorphic genus Branchiobdella seems to be absent in Japan.     

Mechanism of liquefaction of the human ejaculate. I. Changes of the ejaculate proteins.

The spontaneous liquefaction of the human ejaculate in vitro was studied by macroscopic inspection, disc electrophoresis of the ejaculate proteins, free amino acid and nitrogen determinations and by electron microscopy. Liquefaction occurred in three phases. Phase 1 was characterized by the macroscopic solubilization of the gelatinous material and by disappearance of its granular ultrastructure. Zn++, Hg++ and Cu++ completely inhibited the first step of the liquefraction, which could be reversed by Na2EDTA. The factor(s) involved is most probably a proteolytic enzyme(s) which is tightly incorporated into the gelatinous material. In Phase 2 of liquefaction solubilized proteins were degraded to peptides and in Phase 3 these peptides were degraded to amino acids. Factor(s) responsible for these two steps of liquefaction could be removed by washing the gelatinous material.     

Bioelectrorheological model of the cell. 3. Viscoelastic shear deformation of the membrane.

An analytical electromechanical model of a spherical cell exposed to an alternating electric field was used to calculate shear stress generated in the cellular membrane. Shape deformation of Neurospora crassa (slime) spheroplasts was measured. Statistical analysis permitted empirical evaluation of creep of the cellular membrane within the range of infinitesimal stress. Final results were discussed in terms of various rheological models.     

Information from e.x.a.f.s. spectroscopy on the structures of different forms of molybdenum in xanthine oxidase and the catalytic mechanism of the enzyme.

In isolated perfused rat hearts, epidermal growth factor (EGF; 15 nM) increased cellular cyclic AMP (cAMP) content by 9.5-fold. In rat cardiac membranes, EGF also stimulated adenylate cyclase activity in a dose-dependent manner, with maximal stimulation (35% above control) being observed at 10 nM-EGF. Half-maximal stimulation of adenylate cyclase was observed at 40 pM-EGF. Although the beta-adrenergic-receptor antagonist propranolol markedly attenuated the isoprenaline-mediated increase in cAMP content of perfused hearts and stimulation of adenylate cyclase activity, it did not alter the ability of EGF to elevate tissue cAMP content and stimulate adenylate cyclase. The involvement of a guanine-nucleotide-binding protein (G-protein) in the activation of adenylate cyclase by EGF was indicated by the following evidence. First, the EGF-mediated stimulation of adenylate cyclase required the presence of the non-hydrolysable GTP analogue, guanyl-5'-yl-imidodiphosphate (p[NH]ppG). Maximal stimulation was observed in the presence of 10 microM-p[NH]ppG. Secondly, in the presence of 10 microM-p[NH]ppG, the stable GDP analogue guanosine 5'-[beta-thio]diphosphate at a concentration of 10 microM blocked the stimulation of the adenylate cyclase by 1 nM- and 10 nM-EGF. Third, NaF + AlCl3-stimulated adenylate cyclase activity was not altered by EGF. The ability of EGF to stimulate adenylate cyclase was not affected by pertussis-toxin treatment of cardiac membranes. However, in cholera-toxin-treated cardiac membranes, when the adenylate cyclase activity was stimulated by 2-fold, EGF was ineffective. Finally, PMA by itself did not alter the activity of cardiac adenylate cyclase, but abolished the EGF-mediated stimulation of this enzyme activity. The experimental evidence in the present paper demonstrates, for the first time, that EGF stimulates adenylate cyclase in rat cardiac membranes through a stimulatory GTP-binding regulatory protein, and this effect is manifested in elevated cellular cAMP levels in perfused hearts exposed to EGF.     

Postnatal development of the respiratory system of the opossum. II. Electron microscopy of the epithelium and pleura.

At birth, the opossum lung is remarkably primitive and consists of a system of branching airways that end in a number of terminal air chambers. From the newborn through the 10 cm stage of development the conducting portion of the lung predominates. The air chambers, which represent portions of the conducting system modified for respiration, are in a constant state of evolution since they are destined to become part of the expanding bronchial system. The airways are devoid of cilia and goblet cells at birth, and are lined by columnar epithelial cells which contain two types of cytoplasmic granules: an electron-dense form and a heterogeneous form. The latter exhibits an electron-dense core surrounded initially by a large halo of flocculent material. This type of granule is not seen beyond the 8 cm stage. The terminal air chambers of the newborn and later stages are lined type I and type II alveolocytes that appear identical to the alveolocytes lining alveoli in the adult. By the 2.5 cm stage, scattered cilia are present in the trachea and bronchi and bands of smooth muscle have differentiated in relation to bronchial epithelium and to proximal areas of the terminal chambers. Citiated cells are separated by ridges composed of light and dark cells which are without cilia and which contain scattered electron-dence granules. Throughout the postnatal period numerous alveolar macrophages and mast cells are noted in relation to the conducting system and pleura. Differentiation of the pleura also occurs during the postnatal period. In the newborn the pleura is simple squamous mesothelium. Later stages develop a thick connective tissue lamina between the pleural mesothelium and lung parenchyma. A large band of elastin is interposed between the mesothelium and underlying bundles of collagen.     

On the trail of Dr. Fifer.

A gift from a patient drew Hope, BC, family physician Gerd Asche irrevocably into the local medical history of the 1858 Fraser River Gold Rush. Because of his interest in Dr. Max William Fifer, Asche undertook research missions in British Columbia, England and the US, converted his computer room to a research and writing centre, and wrote a biography of his predecessor and colleague. He recounts his experience and the growing satisfaction provided by his interest in medical history.     

Decade of the brain. An agenda for the nineties.

On July 25, 1989, President George Bush, in response to reports written by the National Advisory Councils of the National Institute of Neurological Disorders and Stroke and the National Institute of Mental Health and at the urging of Congress, signed a presidential declaration designating the 1990s to be the "Decade of the Brain" and called on the United States to observe the decade with appropriate activities. At mid-decade, scientific accomplishment has been spectacular; however, both public support and increases in research resources have been minimal. It can be anticipated that scientific progress will continue to be impressive for the remainder of the decade, but many research opportunities will either not be addressed or will be postponed. At mid-decade, the time has come to re-evaluate the research agenda and the public strategy for the remainder of the decade.     

Cascade control of E. coli glutamine synthetase. II. Metabolite regulation of the enzymes in the cascade.

Enzymes and regulatory proteins involved in the cascade control of glutamine synthetase activity of Escherichia coli have been separated from one another and the effects of numerous metabolites on each step in the cascade have been determined. The adenylyl transferase (ATase) -catalyzed adenylylation of glutamine synthetase, which requires the presence of the unmodified form of the regulatory protein PII is enhanced by glutamine and is inhibited by either α-ketoglutarate (α-KG) or the uridylylated form (PII·UMP) of the regulatory protein. PII·UMP and α-KG act synergistically to inhibit this activity. In contrast, the PII·UMP-dependent, ATase-catalyzed deadenylylation of glutamine synthetase requires α-KG and ATP and is inhibited by glutamine or PII and synergistically by glutamine plus PII. The capacity of uridylyl transferase (UTase) to catalyze the uridylylation of PII is dependent on the presence of α-KG and ATP and is inhibited by glutamine. The deuridylylation of PII·UMP by the uridylyl removing enzyme (UR) is enhanced by glutamine but is unaffected by α-KG. However, CMP, UMP, and CoA all inhibit activity at 10^?6m. High concentrations of ATase inhibit both UR and UTase activities, presumably by binding the regulatory protein. Of more than 50 substances that alter the activity of at least one enzyme in the cascade, only α-KG and glutamine affect the activity at every step. This accounts for the observation that glutamine synthetase activity in vivo is very sensitive to the intracellular ratio of α-KG to glutamine.     

E.s.r. study of the effect of oxygen on the irradiation of polyglycine

Radiation damage in polycrystalline polyglycine irradiated under different partial pressures of oxygen has been investigated by e.s.r. spectroscopy. Although the general form of the e.s.r. spectrum remained unaltered as compared to irradiation under vacuum, a decrease in the intensity of the e.s.r. signal was observed. The results were analysed in correlation with Alper's formula. A correspondence between free radical transformation into nonradical species in the presence of oxygen and the biological centres of damage is suggested.     

Comparison of the secretory processes in the parotid and sublingual glands of the mouse. 1. Regulation of the secretory processes.

1. Secretion from the mucous sublingual gland of the mouse has been investigated and compared with the serous parotid gland. The influence of acetylcholine, noradrenalin and adrenalin on the secretion of glycoproteins (e.g. mucins) and proteins (e.g. amylase) from these glands in vitro, and the involvement of cyclic AMP and Ca2+ has been studied. 2. Secretion from the parotid gland could be stimulated by both acetylcholine and the catecholamines. It appears that cyclic AMP plays an important role in the adrenergic secretory process, but not in the cholinergic-induced secretion. In the latter case, exogenous Ca2+ strongly increased the secretion. 3. Mucin secretion from the sublingual gland could be affected by acetylcholine in the presence of exogenous Ca2+. Noradrenalin and adrenalin induced only a slow mucin secretion and, for this secretory process, exogenous Ca2+ is also required. Though cyclic AMP is present in the sublingual gland, no influence on its level could be detected in this gland after stimulation of the adrenergic beta-receptor, whereas, in contrast to the parotid gland, dibutyryl cyclic AMP induced only a slow secretion. Because it was observed that the sublingual gland of the mouse is not innervated sympathetically, it seems reasonable to suppose that the catecholamines stimulate the mucin secretion from this gland via hormonal receptors and not via the adrenergic beta-receptor. 4. The protein secretion from the sublingual gland could be stimulated by both acetylcholine and the catecholamines. An involvement of cyclic AMP in this process was not observed. Addition of exogenous Ca2+ is less important, as was found for the mucin secretion. So it has been concluded that protein and mucin secretion from the sublingual gland are regulated via different pathways.     

Cyanobacterial phycobilisomes. Characterization of the phycobilisomes of Synechococcus sp. 6301.

A procedure is described for the preparation of stable phycobilisomes from the unicellular cyanobacterium Synechococcus sp. 6301 (also known as Anacystis nidulans). Excitation of the phycocyanin in these particles at 580 nm leads to maximum fluorescence emission, from allophycocyanin and allophycocyanin B, at 673 nm. Electron microscopy shows that the phycobilisomes are clusters of rods. The rods are made up of stacks of discs which exhibit the dimensions of short stacks made up primarily of phycocyanin (Eiserling, F. A., and Glazer, A. N. (1974) J. Ultrastruct. Res. 47, 16-25). Loss of the clusters, by dissociation into rods under suitable conditions, is associated with loss of energy transfer as shown by a shift in fluorescence emission maximum to 652 nm. Synechococcus sp. 6301 phycobilisomes were shown to contain five nonpigmented polypeptides in addition to the colored subunits (which carry the covalently bound tetrapyrrole prosthetic groups) of the phycobiliproteins. Evidence is presented to demonstrate that these colorless polypeptides are genuine components of the phycobilisome. The nonpigmented polypeptides represent approximately 12% of the protein of the phycobilisomes; phycocyanin, approximately 75%, and allophycocyanin, approximately 12%. Spectroscopic studies that phycocyanin is in the hexamer form, (alpha beta)6, in intact phycobilisomes, and that the circular dichroism and absorbance of this aggregate are little affected by incorporation into the phycobilisome structure.