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In previous work on rat striated muscle cells a silver-reducing component was found selectively localized at the terminal cistern/transverse tubule system (Tandler and Pellegrino de Iraldi 1989). To further investigate that problem we performed the Hg-Ag argentaffin reaction on a sarcoplasmic reticulum fraction from rat skeletal muscle. Circular profiles corresponding to vesicular structures were found outlined by silver grains. The number of silver "stained" vesicles were less than the total number vesicles stained by conventional procedures. The correlation between argentaffinities in the intact muscle fiber and their subcellular organelles indicated that the Hg-Ag reactive vesicles must be those derived from the terminal cisternae of the sarcoplasmic reticulum. The silver-reducing constituent aggregates in the presence of 1 mM CaCl2 or 0.5 M K cacodylate. The state of aggregation induced by Ca2+ was not affected by incubation with 0.5% Triton X-100 or by 2 mM EDTA, thus suggesting a localization at or near the membrane of the terminal cistern vesicle facing the junctional gap. In Laemmli SDS-acrylamide gels the Hg-Ag reaction stained all proteins in a manner similar to Coomasie blue. It is suggested that the selective histochemical staining is the result of differential reactivities due to steric requirements of the chemical reaction.

Summary. This study reports the presence of a silver-reducing constituent in rat striated muscle fiber located selectively at the level of the terminal cistern/transverse tubule system. It is related to the T tubule network at or near sites that participate in junctions with terminal cisternae, i.e., at both sides of the T tubule in skeletal muscle (triad) and, predominantly, at one side in the ventricle (dyad). Little reactivity is present in the auricle due to the scarcity of those membrane systems. The longitudinal sarcoplasmic reticulum, the sarcolemma, mitochondria and myofibrils are not outlined by the reaction product. Extraction of low molecular weight substances, nucleic acids and lipids did not suppress the chemical reaction. A new argentaffin (Hg-Ag) technique is described. Ethanol or aldehyde fixed muscles were passed to water, postfixed 6-24 h with mercuric acetate (5% w/v in 1% acetic acid), washed with 1% acetic acid and distilled water, stained 12-24 h at 43 ~ C with ammoniacal silver nitrate (60% w/v) and washed in 10% sodium sulfite (three changes) and water. All steps were carried out in darkness. Postfixation with mercuric acetate proved to be essential for immobilizing the argentaffin component without interfering with its strong argentaffinity. The procedure also provides a simple method for tracing the pathway of transversally oriented membrane systems in skeletal and cardiac muscle cells.

The serotonin antigen (5-HT-BSA formaldehyde conjugate) used for obtaining anti-5-HT antibodies was studied to obtain additional data concerning the nature of its immunogen. Dialysis against 0.1 M acetic acid and then against distilled water proved to be the best way of removing 5-HT condensation products not bound to BSA. The hapten has the configuration of a tetrahydro-beta-carboline (THBC) ring structure that is coupled to protein most probably via the carbon(s) ortho to the phenolic hydroxyl group and the indole nitrogen. The cyclic secondary amine of the THBC remained unsubstituted and was not involved in the bridging to BSA. This functional group was effectively blocked by acetylation and was unreactive to glutaraldehyde. On the other hand, in 5-HT conjugates synthesized using glutaraldehyde as the coupling agent, no cyclization to THBC occurred, and the amino groups were blocked. The chemical reactivity of the secondary amino group of the hapten in the synthesized conjugates was compared to the immunoreactivity of 5-HT conjugates formed in tissues. Immunostaining of formaldehyde-fixed serotoninergic neurons of the raphe of rats was suppressed by acetylation and the use of glutaraldehyde as the primary fixative, but the staining was unaffected when glutaraldehyde was reacted with formaldehyde-fixed 5-HT neurons. It is concluded that the cyclic secondary amine of the THBC structure is not conjugated to protein and forms part of the 5-HT-antibody-binding site in immunogens formed in vitro and in tissues.

Transmethylation is known to occur from methionine to ribonucleic acid (RNA) derivatives (Fleissner & Borek, 1962). Most of these methylated derivatives are typical of transfer RNA (Dunn, 1959). In dipteran salivary cells it was possible to distinguish with autoradiography between the transmethylation to RNA and the incorporation of methionine into protein (Sirlin, Jacob & Tandler, 1963). The same approach was used in the present experiments to trace the formation of transfer RNA in amphibian oöcytes.

For localization of pyroantimonate-precipitable cations, rat kidney was fixed by perfusion with a saturated aqueous solution of potassium pyroantimonate (pH about 9.2, without addition of any conventional fixative). A remarkably good preservation of the tissue and cell morphology was obtained as well as a consistent and reproducible localization of the insoluble antimonate salts of magnesium, calcium, and sodium. All proximal and distal tubules and glomeruli were delimited by massive electron-opaque precipitates localized in the basement membrane and, to a lesser extent, in adjacent connective tissue. In the intraglomerular capillaries the antimonate precipitate was encountered in the basement membranes and also between the foot processes. In addition to a more or less uniform distribution in the cytoplasm and between the microvilli of the brush border, antimonate precipitates were found in all cell nuclei, mainly between the masses of condensed chromatin. The mitochondria usually contained a few large antimonate deposits which probably correspond to the so-called "dense granules" observed after conventional fixations.

For localization of pyroantimonate-precipitable cations, mouse testes were fixed with a saturated aqueous solution of potassium pyroantimonate (pH about 9.2, without addition of any conventional fixative), hardened with formaldehyde, and postosmicated. A good preservation of the cell membranes and over-all cell morphology is obtained as well as a consistent and reproducible localization of the insoluble antimonate salts of magnesium, calcium, and sodium. Four sites of prominent antimonate deposits are revealed, besides a more or less uniform distribution of the precipitates. These sites are: (a) In the walls of the seminiferous tubules, localized in two concentric layers corresponding to the inner and outer layers of the tubular wall; (b) Around the blood vessels and adjacent connective tissue; (c) At the area of contact between the Sertoli cell and spermatids, where a double line of precipitate surrounds the head of the mature spermatids; and (d) In the cell nuclei, disposed between regions of the condensed chromatin. The nucleus of mature spermatids did not show any sign of antimonate precipitation. The implications of this inorganic cation distribution are discussed with relation to their anionic counterparts, their localization in other animal and plant tissues, and the possibility that those sites may represent barriers to the free passage of ions.

Lead acetate (3–10%, pH between 4.3 and 7.0, alone or containing 2% glutaraldehyde), when used as fixative, has been demonstrated to produce an intracellular microcrystalline precipitate of lead orthophosphate, Pb5(PO4)3OH (lead hydroxyapatite). This confirms earlier work with the light microscope (6). In interphase cells the nucleoli are sharply delimited by the massive lead phosphate precipitate. Some diffuse precipitate is found in the nucleoplasm; it is always delimited by the nuclear membrane. Nucleolar localization of this orthophosphate pool is not a diffusion artifact; the pool is probably in a loosely bound state and is not retained by conventional fixatives. In maize root cells in advanced mitotic stages the lead phosphate crystals are seen distributed throughout the cytoplasm and also relatively concentrated on the late anaphase-early telophase chromosomes. This pool of inorganic phosphate anions may be involved in the mitotic cycle of chromatin condensation, and it may be partially responsible for the absence of mature ribosomes in the nucleolus through the chelation of divalent cations. It is evident that the siver-reducing component detected in the nucleoli of fixed cells (6) is a completely different substance.

Lead acetate treatment of unfixed cells immobilizes the intracellular water-soluble, inorganic orthophosphate ions as microcrystalline lead hydroxyapatite precipitates (see reference 1). These precipitates have been analyzed with the electron microprobe. A much higher concentration of phosphorus has been found in the nucleoli of maize root tip cells fixed in lead acetate-glutaraldehyde (organic phosphorus plus inorganic orthophosphate), as compared to the nucleoli of roots fixed in glutaraldehyde alone (organic phosphorus). The concentration of the inorganic orthophosphate pool in these nucleoli is three to five times as high as the concentration of the macromolecular organic phosphate. Since nearly all of the latter is in RNA, the concentration of inorganic phosphate in the nucleolus is calculated to be roughly 0.5–0.8 M. About 30%—and up to 50%—of the total cellular inorganic phosphate is accumulated in the nucleolus since the mean concentration per cell is about 10-2 M. In the extranucleolar part of the nucleus the mean concentration was estimated by densitometry to be roughly six times less than in the nucleolus (⩽ 0.1 M), and appears more concentrated in the nucleoplasm than in the condensed chromatin. While there is no direct evidence for the concentration in the cytoplasm, it certainly must be much lower than the mean cellular level (i.e., < 10-2 M) since the nucleus is about 10% of the total cell volume. The implications of this compartmentation in the intact cell are discussed in connection with (A) the availability of orthophosphate ions for the cytoplasm in those processes in which these ions affect the rate of enzymatic reactions, and (B) protein nucleic acid interactions within the nucleus and nucleolus.

Inorganic cations have been localized in several animal and plant tissues by means of the pyroantimonate fixation procedure (1-3) . We demonstrated that an aqueous solution of potassium pyroantimonate enters the cell without disruption of cell membranes and produces electron-opaque cation-antimonate precipitates in a constant and reproducible manner. The results obtained in mouse testis (3) were further extended to the meiotic prophase in spermatogenesis and to Sertoli's cells .' In the present paper we studied the distribution of pyroantimonate-precipitable cations during spermiogenesis . We demonstrate regional differences with respect to the occurrence of the precipitate of the spermatid during evolution .

A. Chromatographische Analysen des Alkali-Hydrolysates von athanolextrahiertenAcetabularia-Zellen ergaben, das grose Mengen noch nicht naher identifizierter, Salkowski-positiver Indole (rotlich) freigesetzt werden. Ihre Menge entspricht ungefahr der des freigesetzten Tryptophans. Die Indolylderivate sind bei der alkalischen Hydrolyse jedoch nicht aus Tryptophan entstanden.

The distribution of pyroantimonate-precipitable inorganic cation was studied inAllium cepa root tips using the direct potassium pyroantimonate fixation procedure. An accumulation of cation-antimonate precipitates was found at the boundary with the cell wall, presumably at the site of the plasmalemma; it extends to numerous infoldings at the cytoplasmic side suggesting a form of pinocytosis. A few large intramitochondrial antimonate deposits were also found and may reflect cation transport into mitochondria. The observed sites of inorganic cation accumulation in the nucleus apparently correlate with its RNA synthetic activity and reside mainly in the nucleolus.

The chemical specificity of the lead acetate reaction (17) has been improved by the use of ammonium acetate and other variants in the cytochemical technique. It has been possible to establish that inorganic phosphate is a normal component of the nucleolus. Whether it is present only in the nucleoli, as suggested by the cytological specificity of the test, or whether it represents only a fraction of the total inorganic phosphate of the cell, remains to be proved.

Earlier reports indicated the presence of significant amounts of inorganic salts in the nucleus. In the present study the possibility that this might be related to the transcription process was tested on seminiferous epithelium of the adult mouse, using potassium pyroantimonate as a fixative. The results indicated that a correlation exists between the inorganic cations comprising the pyroantimonate-precipitable fraction and the RNA synthetic activity. During meiotic prophase an accumulation of cation-antimonate precipitates occurs dispersed through the middle pachytene nuclei, the stage in which RNA synthesis reaches a maximum. At other stages (zygotene to diplotene), where RNA synthesis falls to a low level, that pattern is not seen; cation-antimonate deposits are restricted to a few masses in areas apparently free of chromatin. The condensed sex chromosomes, the heterochromatin of the "basal knobs," the axial elements, and the synaptonemal complexes are devoid of antimonate deposits during the meiotic prophase. The Sertoli cells, active in RNA synthesis in both nucleoplasm and nucleolus, show cation-antimonate deposits at these sites. In the nucleoplasm some "patches" of precipitates appear coincident with clusters of interchromatin granules; in the nucleolus the inorganic cations are mainly located in the fibrillar and/or amorphous areas, whereas relatively few are shown by the granular component. The condensed chromatin bodies associated with the nucleolus were always free of antimonate precipitates. It is suggested that the observed sites of inorganic cation accumulation within the nucleus may at least partially indicate the presence of RNA polymerases, the activity of which is dependent on divalent cations.

The staining technic for the visualization of acid phosphatase is based on the finding that when sections of tissue are placed in a medium containing a phosphone ester, the sites of the enzymatic liberation of phosphate could he determined if lead ions are present to precipitate the phosphate as it is formed. fh deposit of lead phosphate could be converted to an easily visualized black pre(ipitate of lead sulfide (Oomori, 1941). This note describes a technic based on the same principle, but replacing lead by (ol)alt, which precipitates a blue insoluble orthophosphate at loci of phosphatase action. Acetone fixed tissues (1 mm. thick) were washed with distilled water and passed to a I % (obalt a(etate solution for 10 minutes. The tissues were then placed for 12 hours iii it medium (pH 6.0-6.5) containing equal parts 2 % cobalt acetate “stock” solution and 0.2 % sodium-$-glycerophosphate, freshly prepared. The zones where the phosphatase is concentrated develop a deep blue color. It is not possible to use the cobalt acetate technic at pH 5.0 (which is the optimum pH of acid phosphatase) because at this pH precipitation of phosphate is incomplete. Ilowever, cobalt acetate presents the advantage of not precipitating with the phosphoric esters and proteins. For microscopic observation frozen sections 20 to 40 /2 thick were incubated 3 to 6 hours in the substrate medium, rinsed with distilled water, and immersed 15 minutes in either of the two following solutions, prepared just before using: a) Ammonium sulfide-alcohol (1 cc. of concentrated ammonium sulfide in 50 cc. of 70 % alcohol), b) H2S water prepared by bubbling H2S through distilled water. Wash with 70 % alcohol or distilled water respectively, dehyd rate in alcohol and mount in Euparal. The deposit of cobalt phosphate is thus converted to the more easily visualized black precipitate of cobalt sulfide. Simultaneously a control is run by treating tissue with cobalt acetate only, to (heck preformed cobalt-positive substances. Typical blue precipitates are obtained with the root tips of A ilium cepa, rootlets and cotyledons of Pisum sativum, liver and the gray substance of the guinea pig’s spinal cord, among others. Phosphate liberation is inhibited in the presence of sodium fluoride (M/10), by heating of the tissue to 95#{176}C for 30 minutes, by fixation in 10% formalin or by sulfuric acid (0.25 N) treatment for two hours. The enzyme is not inhibited in the presence of iodoacetate (M/ 100) or by exposure of the acetone fixed tissues to 2 citric or acetic acid for 2 to 6 hours. Fixation in Carnoy’s fluid (2#{176}-5#{176}C.)

A flexible, generic model of the Space Station Freedom active thermal control system has been developed which is designed to analyze dynamic interactions of the major subsystems of the ATCS. Models are described for the components of the central thermal bus, the radiator external thermal environment, and the internal thermal control system. Two programs are described which facilitate the development of the integrated ATCS model. The first, SIMRAD, simplifies an external thermal environment model given a desired level of accuracy in integrated model performance. The model reduction technique is shown to reduce model execution time significantly while maintaining the desired accuracy. The second, GENFLU, generates SINDA/FLUINT input code for the evaporator and load interface models and automates the integration of load submodels. The component submodels and integration techniques were used to create an integrated model of the thermal control system for an early assembly flight configuration. The results demonstrate the utility of the integrated model in studying dynamic interactions of the ATCS subsystems.

The article contributes to the development of health-welfare in Austria in the first third of the 20th century. The various relations to such activities of health protection in Germany and the efforts and merits of the Vienna physician, anatomist and social politician Julius Tandler (1869-1936) are accentuated.

A mixture of absolute methanol and acetic anhydride (MA) (5:1, v/v; 24 h at 25 degrees C) which is known to methylate and acetylate--respectively--free carboxyl and amino groups in proteins, was tested for its effectiveness as a blocking agent in glutaraldehyde-fixed mouse testis and skeletal muscle. In young spermatids the staining of the acrosome with either uranyl acetate (UA) or ethanolic phosphotungstic acid (PTA) was completely prevented by prior treatment with MA. Esterification of carboxyl groups with 0.1 N HCl in methanol (24 h at 30 degrees C) eliminated the UA staining without affecting that due to PTA. It is concluded that--COOH groups are responsible for UA binding and that acetylation (of amino groups) prevented PTA binding. MA also abolishes the strong affinity of PTA to the lateral elements of the synaptonemal complex in meiotic chromosomes, the axoneme and fibrous sheath of the spermatid tail and the Z band in skeletal muscle. Reactivity was diminished in nucleoli and remained unaffected in chromatin, the outer coarse fibers of the flagellum and collagen fibrils. Different functional groups may participate in PTA staining. The ultrastructure was well preserved in all cases.

Fur die dringlichen operativen Eingriffe am Urogenitaltrakt kommen vor allem die Behinderungen in der normalen Wegsamkeit der Harnwege in Betracht. Hierzu kommen die Zertrummerungen der Niere mit den eintretenden Blutungen, eventuell noch die Blutungen bei extrauteriner Graviditat. Was zunachst die letzte anbelangt, sei hier kurz darauf verwiesen, das nach der Stellung der richtigen Diagnose der operative Eingriff in der Eroffnung des Cavum peritoraeale mit der daran anschliesenden Entfernung des Fruchtsackes besteht. Profuse Blutungen aus einer geplatzten Tubargraviditat liesen sich auch einschranken, wie dies schon erwahnt wurde, entweder durch die Momburgsche Abschnurung, oder durch die Unterbindung der Arteria hypogastrica.

Aus dem grosen Gebiete der Hernien kommen fur die dringlichen Operationen nur jene Falle in Betracht, in welchen es sich um inkarzerierte Hernien handelt. Obwohl fast alle Formen der Hernien zur Inkarzeration fuhren konnen, soll hier nur die Topographie jener besprochen werden, welche erfahrungsgemas eine gewisse Haufigkeit besitzen. Ich mochte gleich hier anfuhren, das mit der Bezeichnung Hernie insoferne Misbrauch getrieben wird, als eine Reihe von Erscheinungen, welche mit einer Hernie im engeren Sinne des Wortes nichts zu tun haben, mit diesem Namen belegt werden. So spricht man z. B. von Muskelhernien, wenn ein Teil eines Muskels, besonders wahrend der Kontraktion durch einen Schlitz der Faszie vorquillt. Auch Divertikelbildung der Hohlorgane hat man vielfach als Hernien bezeichnet. Wir wollen unter Hernie im strengen Sinne des Wortes nur den Austritt von Eingeweidestucken des Cavum peritonaeale unter gleichzeitiger Vorstulpung des daselbst befindlichen Peritonaeums und der daruber gelegenen Schichten verstehen. Mechanisch sind die Hernien der Ausdruck der Differenz zwischen der Hohe des Abdominaldruckes und der Widerstandsfahigkeit der Abdominal wand an der betreffenden Stelle. Uberall dort, wo aus irgendeinem Grunde, sei er traumatischen Ursprunges oder in der Textur der Wand gelegen, der Wandwiderstand der Hohe des Abdominaldruckes nicht gewachsen ist, kommt es zur Bildung einer Hernie. Nach diesem Prinzip konnen wir zwei Formen der Hernien unterscheiden.

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Krause operiert bei aufrecht sitzendem Patienten und last sich von einem Assistenten den Schadel in dieser Stellung halten. Zweckmasiger erscheint uns, wie auch anderen Chirurgen (Frazier, Borchardt) die liegende Stellung mit Hilfe eines eigenen Apparates, wie ihn Cushing angegeben hat. Der Patient liegt am Bauch, der Kopf ruht mit Stirne und Wange auf einer halbmondformigen Stutze, welche am oberen Ende des Operationstisches aufgestellt wird. Damit die Atmung bei dem am Bauch liegenden Patienten nicht gehindert wird, wird die Brust des Patienten auf Binden gelagert, welche zwischen zwei Achselstutzen ausgespannt werden.

Die Hirnpunktion wird teils zu diagnostischen, teils zu therapeutischen Zwecken ausgefuhrt.

A new type of cytoplasmic structure is described in the root tips ofAllium cepa and several other genera. They are very probable composed of insoluble phosphate salts and as such can be demonstrated at the light and electron microscope level by conversion into its lead salt. They appear to be formed by coalescence of electron dense “vesicles” which have granular phosphate deposits at their periphery.

Fur die Operationen an der Konvexitat des Groshirns kommen folgende Regionen in Betracht: 1. die motorische Region, 2. das Stirnhirn, 3. der Parietal- und Hinterhauptlappen und 4. der Schlafelappen.

Lead acetate treatment of unfixed cells immobilizes the intracellular water-soluble, inorganic orthophosphate ions as microcrystalline lead hydroxyapatite precipitates (see reference 1). These precipitates have been analyzed with the electron microprobe. A much higher concentration of phosphorus has been found in the nucleoli of maize root tip cells fixed in lead acetate-glutaraldehyde (organic phosphorus plus inorganic orthophosphate), as compared to the nucleoli of roots fixed in glutaraldehyde alone (organic phosphorus). The concentration of the inorganic orthophosphate pool in these nucleoli is three to five times as high as the concentration of the macromolecular organic phosphate. Since nearly all of the latter is in RNA, the concentration of inorganic phosphate in the nucleolus is calculated to be roughly 0.5–0.8 M. About 30%—and up to 50%—of the total cellular inorganic phosphate is accumulated in the nucleolus since the mean concentration per cell is about 10-2 M. In the extranucleolar part of the nucleus the mean concentration was estimated by densitometry to be roughly six times less than in the nucleolus (⩽ 0.1 M), and appears more concentrated in the nucleoplasm than in the condensed chromatin. While there is no direct evidence for the concentration in the cytoplasm, it certainly must be much lower than the mean cellular level (i.e., < 10-2 M) since the nucleus is about 10% of the total cell volume. The implications of this compartmentation in the intact cell are discussed in connection with (A) the availability of orthophosphate ions for the cytoplasm in those processes in which these ions affect the rate of enzymatic reactions, and (B) protein nucleic acid interactions within the nucleus and nucleolus.

Die Punktion wird ausgefuhrt: 1. Zu diagnostischen Zwecken: Untersuchung des Liquor auf Blut, Entzundungsprodukte, Bakterien. 2. Zu therapeutischen Zwecken zur Druckentlastung oder Einverleibung von Medikamenten in den Duralsack. 3. Zur Einleitung der Ruckenmarksanasthesie.

For localization of pyroantimonate-precipitable cations, mouse testes were fixed with a saturated aqueous solution of potassium pyroantimonate (pH about 9.2, without addition of any conventional fixative), hardened with formaldehyde, and postosmicated. A good preservation of the cell membranes and over-all cell morphology is obtained as well as a consistent and reproducible localization of the insoluble antimonate salts of magnesium, calcium, and sodium. Four sites of prominent antimonate deposits are revealed, besides a more or less uniform distribution of the precipitates. These sites are: (a) In the walls of the seminiferous tubules, localized in two concentric layers corresponding to the inner and outer layers of the tubular wall; (b) Around the blood vessels and adjacent connective tissue; (c) At the area of contact between the Sertoli cell and spermatids, where a double line of precipitate surrounds the head of the mature spermatids; and (d) In the cell nuclei, disposed between regions of the condensed chromatin. The nucleus of mature spermatids did not show any sign of antimonate precipitation. The implications of this inorganic cation distribution are discussed with relation to their anionic counterparts, their localization in other animal and plant tissues, and the possibility that those sites may represent barriers to the free passage of ions.

Die Eroffnung der knochernen Schadelkapsel kann auf zweierlei Weise erfolgen: 1. mit dauernder Wegnahme des Knochens, 2. mit temporarer Wegnahme des Knochens (osteoplastische Operation).