[Anomalies of the inferior vena cava (author's transl)].

After a detailed account of the developmental anatomy of the inferior vena cava, which arises from various segments, an attempt is made at a systematic classification of anomalies of the inferior vena cava. For this purpose, the angiographic appearances and available literature have been considered. The following anomalies have been distinguished: 1. left-sided position, 2. duplication, 3. anomalous insertion into the left atrium, 4. agenesis and 5. stenosis and occlusion.


[Renal cystography--a diagnostic and therapeutic procedure (author's transl)].

Pre-operatively, renal cysts can be distinguished with certainty from solid tumours only by their direct radiological demonstration; this is best done by double contrast methods after percutaneous puncture (renal cystography). By introducing a small amount of oily contrast medium, the cyst may be cured at the same time. The method is described and the results in different patients are reported. This combined diagnostic-therapeutic procedure is proposed as an alternative to surgery of renal cysts and is recommended as the method of choice in old patients and those carrying an increased risk.


[Indirect lymphography of the retrosternal lymphatics and of the thoracic duct (author's transl)].

Successful demonstration of the retrosternal lymphatics and of the thoracic duct after infradiaphragmatic injection is reported, using a contrast medium developed by the author. The lymphatics opacify after 45 minutes. After fourteen days the substance can no longer be seen radiogically. Following some preliminary investigations, it is proposed to try the method in humans. The importance of the lymphatic pathways in medial carcinomas of the breast and in the movement of carcinoma cells from the peritoneal cavity and the liver is stressed.


[The mechanism of contrast filling of the liver following lymphography (author's transl)].

Contrast filling of the liver after lymphography is described in one patient. The authors agree with the literature which indicates the significance of lymphatic blocks and caval obstruction. A splenoportogram was demonstrated during the performance of a cavogram and in this way direct contrast filling of the liver is demonstrated for the first time.


[Extra-thoracic sarcoidosis of the retroperitoneal lymph nodes and of the spleen with ascites. Comments on the problem of lymphatic obstruction (author's transl)].

Sarcoidosis without radiologically demonstrable intrathoracic changes is described, in which there was involvement of the retroperitoneal lymph nodes and of the spleen, together with ascites. Contrary to descriptions in the literature of the lymphographic findings in sarcoidosis, there is evidence of lymphatic obstruction and at operation fluid was found in the peritoneal cavity. The relationship between the lymphatic obstruction which was associated with low-grade peritoneal involvement and the increased amount of peritoneal fluid is discussed.


[A modified method for labeling erythrocytes with 99mTc (author's transl)].

A modified method for labeling erythrocytes with 99mTc-sodium pertechnetate for spleenic scintigraphy is described. In vitro investigations have shown that by omitting the customary process of washing and by simultaneous alteration of the erythrocytes during the tin incubation period, the duration of erythrocyte labeling can be considerably reduced. A yield of 96 of 98% can be achieved. The effect of the tin on the erythrocytes is further investigated. The effect of the incubation period and temperature, of the plasma in the erythrocyte suspension, of stirring and agitation, of the dispersion liquid and tin concentration were studied. The method proves satisfactory in 56 controlled spleenic scintigrams.


[Clinical evaluation of cardiovascular reactions to renal contrast media using an impedance method (author's transl)].

The hemodynamic changes during intravenous urography were studied in 120 non-selected patients (60 patients with bolus injection, 60 with infusion: 30 of each group with methylglucamine ioxitalamate, while the other 30 with sodium methylglucamine diatrizoate). There was a uniform statistically significant increase in heart rate (p less than 10(-9) as well as in cardiac output (p less than 10(-9)) (measured by means of a non-invasive impedance method). The observed changes were more marked following bolus injection than infusion. With the application of methylglucamine ioxitalamate the increase in heart rate, but not in cardiac output, was significantly less (p less than 0,01) than with that of sodium methylglucamine diatrizoate.


Complications by translumbar aortography.

The complications resulting from different methods of translumbar aortography were evaluated by checking 1192 examinations performed with 1) a cannula, 2) a straight catheter, and 3) a curved catheter. The latter method was found to be far more safe than the other two. According to the complication rates found in the literature the curved catheter translumbar method is also safer than the transaxillar and comparable to transfemoral aortography by the Seldinger method.


[New aspects on the mode of action of cardiac glycosides].

A dissociation of the therapeutic from the toxic effects of cardiac glycosides has repeatedly been described. Whereas it is generally accepted that the toxic effects of cardiac glycosides are based on an inhibition of the Na+-K+-ATPase, the mechanism of action of therapeutic concentrations of cardiac glycosides still remains uncertain. To test the hypothesis, that cardiac glycosides might be transported into a distinct compartment of the myocardium with the Na+-K-ATPase acting as a carrier, the interaction of some inhibitors of this enzyme (digitoxin, dihydroouabain, cassaine, N-ethylmaleimide, p-hydroxy-mercuribenzoate, ethacrynic acid, spironolactone) with ouabain was studied at different levels of cardiac glycoside actions: Myocardial function, cardiac uptake and subcellular distribution and binding to the Na+-K+-ATPase. The following results were obtained: All cardioactive drugs (ethacrynic acid and spironolactone showed no such effects) reduced dose-dependently the inotropic action of ouabain and in high concentrations increased its toxicity. The same drugs inhibited dose-dependently the cardiac uptake of ouabain without affecting the subcellular distribution pattern of ouabain. The binding of ouabain to the Na+-K+-ATPase was influenced in a similar way by these drugs, showing a competitive type of interaction with digitoxin, dihydroouabain and cassaine and a non-competitive mechanism with N-ethylmaleimide and p-hydroxymercuribenzoate. These results support the concept of a cardiac glycoside-ATPase interaction as a basis for the therapeutic action of these drugs. This may be explained either by a direct influence of cardiac glycosides on the ATPase activity and/or by a carrier mediated cardiac glycoside-transport into a distinct compartment of the myocardial cell.


[Liver tumors. Laparoscopic aspects].

A review is given on laparoscopic possibilities in the differentiation of tumours of the liver. Benign lesions are rare and usually discovered by chance. Primary cancer of the liver can be verified ante finem in a maximum of 40% of the cases. Characteristic is the endoscopic picture in the case of haemangiosarcoma after arsenic, Thorotrast, vinyl chloride (marked capsular fibrosis, net-like marking, "restless" surface of theliver). All the tumours need to be confirmed histologically. A tumour-like appearance is presented by echinococcus alveolaris and coarse-nodular granulomatous changes (sarcoidosis, lymphogranulomatosis, etc.).


[Considerations regarding the influence of intrauterine and early postnatal diseases and nutritional deficiencies on immunity and disease epidemiology (author's transl)].

Malnutrition, infectious and toxic stress, hormonal and enzymatic deficiencies as well as graft versus host reactions during the last trimester of pregnancy and during the first six months of life lead to persistent depressions of cell mediated immunity. The subsequent imbalance between the cell mediated and humoral system of immunity leads to differences in disease prevalence in poor and rich populations. Particularly leprosy, tuberculosis, viral disease as for instance frequently fatal measles and diseases due to complexes between humoral antibody and bacterial components as for example acute rheumatic fever occur with increased frequency in B (+) T (-) populations. Desturbances of immune surveillance due to suppression of specific cell mediated immune function leads to an increased frequency of neoplasia, particularly B-cell lymphoma and gastrointestinal tumors. Populations in which the T-cell system can mature without interference show a trend towards diseases in which excessive T-cell response plays a major role, as for instance rheumatoid arthritis, Sjögren syndrome, terminal ileitis, autoimmune angiopathies, multiple sclerosis and possibly also disseminated lupus erythematodes.


A micromethod for the activation of human blood lymphocytes in vitro.

A micro culture system is described in which 2.5 X 10(4) human blood lymphocytes in aliquots of 100 mul are stimulated by PHA, Pokeweed, "Varidase" antigen, allogeneic small lymphocytes or mitomycin-C-treated allogeneic LCL cells. Careful regulation of the pH by a combination of bicarbonate and MOPS buffers seems to be important for detecting a response to weak stimuli. High and reproducible levels of activation by powerful stimuli (PHA and LCL cells) can be recorded from even smaller cultures (10(4) responding cells in 40 mul aliquots). The technique allows large numbers of replicate cultures to be set up from a single blood sample so that the time course and/or dose-response relationships can be examined for a range of differen mitogens.


I. Evidence that lymphotoxin activity involves both cytotoxic and stimulating factors.

This paper describes experiments undertaken to determine more uniform conditions for the generation of cytotoxic lymphokine--'lymphotoxin' (LT) in differently stimulated rat lymphocyte cultures, and to compare the sensitivity of different test systems for detecting rat LT activity in vitro. Wistar rat lymphoid cells were activated by culture with phytohaemagglutinin (PHA) and mixed lymphocyte cultures (MLC) with semi-allogeneic (Wistar x August) F1 lymphoid cells. Lymphocyte supernatants harvested between 1 and 8 days were tested for their effect on metabolism and viability of cultured mouse fibroblasts (L-929 cells) by four methods: (i) inhibition of [14C]leucine incorporation; (ii) total and viable cell counts in test tube cultures ('macro test'); (iii) viable cell counts in microtitre plates ('microtest'); and (iv) chromium (51Cr) release from chromated L-cell monolayers. Cytotoxic effects on target cells of lymphocyte supernatants were evident after 3 days of PHA stimulation and 6 days of mixed lymphocyte culture, and the most sensitive indication of cytotoxic activity provided by inhibition of amino-acid incorporation and by loss of viable L cells from monolayers in tube cultures. In dilutions greater than 1:16-1:32 both cytotoxic supernatants exhibited a stimulating effect on target-cell proliferation. Stimulation of L cells growth was also observed when monolayers were exposed for 24 h to 'early' (24 h) PHA undiluted supernatants. At a later time of exposure to these supernatants a considerable loss of total and viable cells in the monolayers was evident. The results indicated that both cytotoxic and growth-stimulating lymphokines could be generated during activation of rat lymphocytes. A hypothesis is suggested whereby 'lymphotoxin' activity in vitro arises from the sequential effects of stimulating and cytotoxic lymphokine, and whereby the balance of these effects in vivo might determine the response of fibroblasts involved in reactions of chronic allergic inflammation.


Biogenic amines and affective disorders. A critical analysis.

The evidence linking biogenic amines and affective disorders is critically reviewed. Surveyed are studies on the level of biogenic amines and their metabolites in the brain, blood, urine and cerebrospinal fluid of patients with affective conditions; the effects of biogenic amine precursors, depletors and blockers on affective states and the action of present methods of treatment of these disorders on the level of biogenic amines. Reference is also made to the existence of various disease states where abnormalities of biogenic amines exist in the absence of affective disorders. The review fails to uncover convincing evidence that affective disorders are related to abnormal levels of biogenic amines. Reasons are outlined why the "catecholamine hypothesis" and related theories can neither be proven nor disproven with the available techniques.


[Hemosiderin pigmentation in Sezáry's syndrome].

A 75 year old woman suffering from Sezary syndrome with edemas and excessive pigmentations is presented. In previous observations, pigmentation was caused by melanosis. In our patient it is caused by hemosiderosis. The origin of the pigmentation by a purpura-pigmentosa-progressiva-like mechanism or by the tumor disease itself is discussed.


[Trichosporon capitatum as pathogen of dermatomycoses].

Three cases of dermatomcycosis are reported, caused by Trichosporon capitatum, a yeast-like organism, belonging to genus Trichosporon of the Cryptococcaceae, which till now was isolated out of dermatological material only in a few cases. One of the lesions, characterized as "leuconychia mycotica", is a problematic task with regard to the treatment, for it is a professional infection in an offset-printer. A glycerol containing fluid for wiping his printing machine was detected as reservoir of yeastlike agents and continuously scattering infection cause. The pathogenic role of Trichosporon capitatum is discussed with regard to the present literature and to the three reported cases.


[Ultrastructure and microflora in follicles and comedones].

The resident flora of skin surface, sebaceous follicles, and acne comedones: Pityrosporum, micrococci, and corynebacterium (propionibacterium) acnes is described. The nomenclature, topography, incidence, pathogenic role, and especially the ultrastructural morphology of these organisms are presented.


Effect of calcium on creatine kinase activity of cerebellum.

The effect of Ca++ ions on the histochemical activity of creatine kinase (CK) was investigated in striated muscle and cerebellum of the rat. The intensity and pattern of CK activity was unchanged in the striated muscle when Ca++ was present in the incubation medium instead of Mg++. In the cerebellum, however, Ca++ inhibited the enzymatic activity except in the Purkinje cells.


[Mersilene mesh in reconstruction of the osseous walls of the pneumatised skull (author's transl)].

The use of Mersilene mesh in reconstruction of the stable walls anterior to the air-containing sinus cavities is emphasized. The basic advantage of the technique employed is the need for tissue covering on only one side of the mesh. Subsequent penetration of the mesh by connective tissue with secondary epithelialization from adjacent mucosa develops a stable wall, even in the presence of local infection. The possibility for good fixation to surrounding bone, immediate availability as well as good cosmetic and functional results recommend its application to selected patients.


Analogs and conversion products of streptozotocin.

Reduction of the nitroso group in streptozotocin (1a) has led to cyclized products rather than a semicarbazide (1c). Some analogs (1e, 9a, 9b, 9c and 9d) of streptozotocin in which the nitroso group was replaced by other groups have been prepared.


Activation of calcium transport in skeletal muscle sarcoplasmic reticulum by monovalent cations.

The rates of calcium transport and Ca2+-dependent ATP hydrolysis by rabbit skeletal muscle sarcoplasmic reticulum were stimulated by monovalent cations. The rate of decomposition of phosphoprotein intermediate of the Ca2+-dependent ATPase of sarcoplasmic reticulum was also increased by these ions to an extent that is sufficient to account for the stimulation of calcium transport and Ca2+-dependent ATPase activity. The order of effectiveness of monovalent cations tested at saturating concentrations in increasing rate of phosphoprotein decomposition is: K+, Na+ greater than Rb+, NH4+ greater than Cs+ greater than Li+, choline+, Tris+.


Chemical cross-linking studies of chloroplast coupling factor 1.

Cross-linking reagents have been used to link covalently adjacent subunits of solubilized spinach chloroplast coupling factor 1, which is a latent ATPase. 1,5-Difluoro-2,4-dinitrobenzene, dimethyl-3,3'-dithiobispropionimidate, and dimethylsuberimidate are able to form bridges of 3 to 11 A between amino groups, and hydrogen peroxide and the o-phenanthroline-cupric ion complex catalyze the oxidation of intrinsic sulfhydryl groups. The five individual subunit bands (alpha, beta, gamma, delta, and epsilon) and several new aggregate bands can be separated by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The same four fastest moving aggregate bands, as characterized by their mobilities, migrate more slowly than the heaviest subunit band and appear with all of the cross-linkers employed. The subunit composition of the aggregate bands has been determined through the use of the reversible cross-linkers, dimethyldithiobispropionimidate, (o-phenanthroline)2Cu(II), and H2O2, and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis in which aggregates are separated in the first dimension, the disulfide cross-links are cleaved, and the individual subunits present in the aggregates are separated in the second dimension. The subunits are detected by Coomassie brilliant blue staining and by labeling some of the sulfhydryl groups of the gamma and epsilon subunits with radioactive N-ethylmaleimide. The results obtained indicate that the alpha and beta subunits can cross-link directly with each of the other subunits, that two beta subunits are adjacent, and that gamma epsilon, gamma epsilon 2, alpha delta, and beta delta aggregates are present. A minimal subunit stoichiometry consistent with these results is alpha 2 beta 2 gamma delta epsilon 2. A possible structural model of the coupling factor is derived from the data. Similar, but less extensive, experiments have been carried out with the heat-activated coupling factor (which is an ATPase); no differences in the spatial arrangement of subunits are detected from the two-dimensional gel electrophoresis analysis of the cross-linked aggregates.


Removal of "tightly bound" nucleotides from soluble mitochondrial adenosine triphosphatase (F1).

Soluble mitochondrial ATPase (F1) from beef heart prepared in this laboratory contained approximately 1.8 mol of ADP and 0 mol of ATP/mol of F1 which were not removed by repeated precipitation of the enzyme with ammonium sulfate solution or by gel filtration in low ionic strength buffer containing EDTA. This enzyme had full coupling activity. Treatment of the enzyme with trypsin (5 mug/mg of F1 for 3 min) reduced the "tightly bound" ADP to zero, abolished coupling activity, but had no effect on the ATPase activity, stability, or membrane-binding capability of the F1. When the trypsin concentration was varied between 0 and 5 mug/mg of F1, tightly bound ADP was removed to varying degrees, and a correlation was seen between amount of residual tightly bound ADP and residual coupling activity. Gel filtration of the native F1 in high ionic strength buffer containing EDTA also caused complete loss of tightly bound ADP and coupling ability, whereas ATPase activity, stability, and membrane-binding capability were retained. The ADP-depleted F1 preparations were unable to rebind normal amounts of ADP or any ATP in simple reloading experiments. The results strongly suggest that tightly bound ADP is required for ATP synthesis and for energy-coupled ATP hydrolysis on F1. The results also suggest that ATP synthesis and energy-linked ATP hydrolysis rather than involving one nucleotide binding site on F1, involve a series or "cluster" of sites. The ATP hydrolysis site may represent one component of this cluster. The results show that nonenergy-coupled ATP hydrolysis on F1 can occur in the absence of tightly bound ADP or ATP.


Removal of "tightly bound" nucleotides from phosphorylating submitochondrial particles.

Phosphorylating submitochondrial particles from beef heart (ETPH) prepared here contained about 2.4 nmol of ATP and 1.9 nmol of ADP/mg of protein after repeated washing of the particles. Essentially all of the "tightly bound " ATP and ADP was removed by trypsin treatment. The trypsin-treated ETPH had increased ATPase activity, undiminished NADH oxidase and succinate oxidase activity, but energy-coupling activity (ATP-driven reversed electron transfer) was abolished. Removal of half the ATP and ADP occurred at low levels of trypsin and was associated with loss of half of the coupling activity. Gel filtration of ETPH in high ionic strength buffer also removed ADP and ATP from the particles, resulting in loss of energy-coupling activity, while ATPase activity was increased. The results support the contention that the tightly bound ADP is essential in energy coupling in mitochondria. Tightly bound ATP may also play an essential role.


Exposure of free amino groups in the coupling factor of energized spinach chloroplasts.

Spinach chloroplasts were incubated in the dark with methyl acetimidate in order to amidinate and thereby protect free amino groups. An energy-dependent attack on the coupling factor (CF1) of these amidinated chloroplasts by trinitrobenzenesulfonate was apparent in a second reaction, as long as the reagent was applied in the light or after an acid-base transition. Trinitrophenyl residues approached one each on the alpha and beta subunits, and two to three on the gamma subunit polypeptide of CF1. Accompanying trinitrophenylation was an inhibition of the ATPase activity of CF1 due to a major decrease in the affinity for ATP; however, neither the maximal ATPase rate nor the ability of the protein to serve as a coupling factor for EDTA-extracted chloroplasts was affected. Trinitrophenylation and consequent inhibition of ATPase were 50% prevented by the presence of phosphate, or ADP, or ATP during exposure to trinitrobenzenesulfonate. The protective effects of adenylates were additive with those of phosphate. The ratio of trinitrophenyl groups on the three subunits concerned was the same whether phosphate or ATP was providing 50% protection, or whether neither was present. It is inferred that a conformational change occurs in the amidinated coupling factor when a proton activity gradient is placed across the membranes, and effective ligands tend to prevent the resulting exposure of free amino groups from a previously hidden location.


Restoration of calcium transport in the trypsin-treated (Ca+ + Mg2+)-dependent adenosine triphosphatase of sarcoplasmic reticulum exposed th sodium dodecyl sulfate.

When sarcoplasmic reticulum vesicles are exposed to trypsin for 1 min the adenosine triphosphatase (Mr = 102,000) is cleaved to fragments of Mr = 45,000 and 55,000. The purified ATPase, containing both fragments, transports Ca2+ when incorporated into vesicles containing excess phospholipid. The two fragments can only be dissociated in solutions containing 1% sodium dodecyl sulfate (SDS). Ca2+ transport activity is restored in SDS-dissociated preparations in a series of steps involving dilution with 5 volumes of 5% phospholipids in 0.75% sodium cholate, incubation in ice for 30 min, and passage through an anion exchange column. Vesicles formed in this procedure regain high Ca2+ transport activity if they are incubated in SDS solution at 24 degrees for less than 20 min. However, the extent of renaturation diminishes if the vesicles are incubated for longer periods and little acitivity is recovered in vesicles incubated longer than 60 min at 24 degrees.


Behavior of the Ca2+ transport sites linked with the phosphorylation reaction of ATPase purified from the sarcoplasmic reticulum.

Both the formation and decay of phosphorylated ATPase, and Ca2+ release from and rebinding to transport sites alpha sites) on the enzyme have been investigated in the pre- and post-steady states by means of rapid mixing acid quench technique and a stopped flow technique, respectively. At a relatively high concentration of enzyme (10 muM protein) and a low concentration of free Ca2+ (1.42 muM), rapid changes in the Ca2+ affinity of the transport sites could be monitored with the use of a Ca2+ indicator dye, Arsenazo III. As the enzyme becomes phosphorylated, Ca2+ is released. In the early stages, the ratio of Ca2+ released to acid-stable phosphoenzyme is less than 2, while at the maximum of Ca2+ release, which occurs later than the maximum phosphorylation, it is greater than 2. Assuming that phosphorylation of the enzyme releases both Ca2+ bound to it (Ikemoto, N. (1975) J. Biol. Chem. 250, 7219), these data are consistent with the sequential formation of two acid-stable intermediates differing in Ca2+ affinity and a third acid-labile phosphorylated species of low Ca2+ affinity. The changes in Ca2+ affinity are probably related to changes in the structure of the ATPase transport protein that are involved in the Ca2+ translocation in the intact sarcoplasmic reticulum. As the ATP is being used up, Ca2+ rebinding occurs concomitantly with the decay of phosphoenzyme. The comparison of the time courses of Ca2+ rebinding with that of dephosphorylation in the post-steady state suggests that interactions among phosphorylated and nonphosphorylated ATPase molecular may also be involved in the Ca2+ affinity changes.


5-bromodeoxyuridine may alter the differentiative program of the embryonic pancreas.

The thymidine analog, 5-bromodeoxyuridine (BrdU), inhibits the differentiation of the acinar cells of the embryonic rat pancreas, while having little effect on the growth of the tissue. The BrdU-treated pancreas contains elevated alkaline phosphatase and carbonic anhydrase activities, and, unlike the normal pancreas, contains numerous extracellular fluid-filled vacuoles, surrounded by ductlike cells. Both alkaline phosphatase and carbonic anhydrase activities are located preferentially in the ductlike cells lining the vacuoles. The biochemical, morphological, and functional features of these epithelial cells are therefore characteristic of the normal pancreatic duct cell. Thus, in the exocrine pancreas, BrdU seems to alter the normal program of differentiation by favoring the functional duct cells while inhibiting the differentiation of acinar cells.


Isolation and characterization of subcellular membranes of Entamoeba invadens.

A method is described for the isolation of subcellular membranes of Entamoeba invadens. Plasma membranes were obtained by rate centrifugation followed by isopycnic centrifugation on a sucrose gradient. Intact phagolysosomes floated in a 10% sucrose solution providing a simple technique for isolation. Phagolysosomal membranes were collected by isopycnic centrifugation, after lysis of the phagolysosomes. Microsomes were obtained by differential centrifugation. Membrane fractions were examined by electron microscopy, and the contamination of each fraction was determined with marker enzymes. Mg2+-ATPase is associated with the plasma membrane. Acid phosphatase (beta-glycerophosphate) was associated mainly with phagolysosmal membranes. Plasma membranes also contained acid phosphatase activity which hydrolyzes p-nitrophenylphosphate but not beta-glycerophosphate. The localization of the two phosphatases was confirmed cytochemically. Isolated plasma membranes were contaminated with phagolysosomal membranes (15%) and with microsomes (25%). No more than 5% of the phagolysosomal membrane fraction consisted of plasma membranes. Contamination of the microsomes by plasma and phagolysosomal membranes was 10% and 7%, respectively. Plasma membranes and phagolysosomal membranes had a high ratio of cholesterol to phospholipid (0.93 and 1.05 mumol/mumol, respectively). Microsomes were relatively poor in cholesterol (0.39 mumol/mumol). Microsomes, plasma, and phagolysosomal membranes contained increasing amounts of spingolipids (12%, 17%, and 28%). Phagolysosomal membranes had a high percentage of phosphatidylserine but little phosphatidylcholine. Microsomes were rich in phosphatidylcholine (45%). Differences in phospholipid composition between plasma and phagolysosomal membranes are discussed in view of the phagocytic process.


The effect of antidynein 1 serum on the movement of reactivated sea urchin sperm.

Rabbit antiserum prepared against an ATPase-containing tryptic fragment of dynein by Ogawa and Mohri (J. Biol. Chem. 250: 6476-6483) specifically inhibited the ATPase activity of dynein 1 and not that of dynein 2. Varying amounts of this antidynein 1 serum were added to demembranated sperm while they were swimming in reactivating solution containing 1 mM ATP. The sperm continued to form regularly propagated flagellar bending waves, but the beat frequency decreased gradually with time, the greater part of the change occurring in the first 15 min. The beat frequency after 1 h was a function of the amount of antiserum used, and could be as low as 1 Hz. The waveforms of the treated sperm resembled those of normal reactivated sperm except that the bend angles of both the principal and reverse bends were larger in the proximal portion of flagellum. The ATPase activity and corresponding beat frequency of sperm which had been pretreated with varying amounts of antidynein 1 serum for 15 min at 0 degrees C and then diluted were both decreased as a function of the amount of antiserum added, the ATPase activity of homogenized, nonmotile sperm also decreased upon pretreatment with antiserum, but the percentage decrease was less than for motile sperm. For moderate to low concentrations of antiserum, the rates of reaction with motile and with rigor sperm were almost identical. The overall results suggest that antidynein 1 inhibits the functioning of the dynein arms, probably by blocking the ATPase sites of the dynein 1.


Glucose and ATP levels in pancreatic islet tissue of normal and diabetic rats.

It has been suggested that the hyperglucagonemia observed in diabetic animals and man may be due to an impairment of glucose uptake and metabolism by the alpha-cells resulting in a decreased production of ATP. To test this hypothesis glucose, ATP, glucagon, and insulin were measured in pancreatic islets of normal and alloxan or streptozotocin diabetic rats. Two experimental approaches were used. In the first, the pancreas was perfused in vitro for assessing insulin and glucagon release due to 10 mM amino acids with and without 5 mM glucose. These perfusions were performed in the presence and absence of insulin. After perfusion, the pancreas was frozen and processed for analysis of islet glucose, ATP, insulin, and glucagon content. The second approach was to investigate the islet sucrose, urea, and glucose spaces together with ATP, insulin, and glucagon content in vivo in normal and in insulin-treated and untreated streptozotocin diabetic rats. Perfusion of the pancreas in vitro with 5 mM glucose resulted in higher glucose content of normal islets than in alloxan and streptozotocin diabetic islets. Similarly in the in vivo studies, the intracellular glucose space of the streptozotocin diabetic islets was 30% the value found in normals. In the in vivo experiments, despite the relatively small intracellular glucose space of alpha-cell islets, the ATP content of these islets was only 15-20% lower than the ATP content of normal islets. In the in vitro experiments, perfusion with glucose resulted in ATP contents of alpha-cell islets and of normal mixed alpha-beta-cell islets which were indistinguishable. However, the ATP content of alpha-cell islets was maintained for prolonged periods in the absence of glucose in contrast to mixed islets, composed primarily of beta-cells, in which the ATP level decreased by 45% when glucose-free medium was perfused for sustained periods. Finally, insulin infused in high concentrations or administered to the diabetic animal had no effect on the glucose spaces or the ATP contents of normal or alpha-cell islets. It can be calculated that in vivo the intracellular glucose level of islets from streptozotocin treated rats is approximately 15 mM. Since in normals an extracellular glucose concentration of this magnitude inhibits stimulated glucagon release completely, it would seem unlikely that a lack of intracellular glucose is the cause of the apparent glucose "blindness" of the alpha-cells in diabetes. In fact, in perfusion studies as little as 2.5 mM free intracellular glucose was sufficient to suppress glucagon secretion from diabetic alpha-cells. The results of the ATP measurements clearly eliminate a possible energy deficit of diabetic alpha-cells as cause of the apparent glucose resistance of alpha-cells.


The Sézary syndrome: a malignant proliferation of helper T cells.

The Sézary syndrome is a frequently lethal disease characterized by circulating malignant cells of thymus-derived (T)-cell origin. The capacity of circulating malignant lymphocytes from patients with this syndrome to synthesize immunoglobulins and to function as helper or suppressor cells regulating immunoglobulin synthesis by bone marrow-derived (B) lymphocytes was determined. Peripheral blood lymphocytes from normal individuals had geometric mean immunoglobulin synthetic rates of 4,910 ng for IgM, 1,270 ng for IgA, and 1,625 ng for IgG per 2 X 10(6) cells in culture with pokeweed mitogen for 7 days. Purified normal B cells had geometric mean synthetic rates of 198 ng for IgM, 145 ng for IgA, and 102 ng for IgG. Leukemic cells from patients with the Sézary syndrome produced essentially no immunoglobulins. Adding normal T cells to normal B cells restored their immunoglobin producing capacity. Leukemic cells from four of five patients tested had a similar capacity to help immunoglobulin synthesis by purified normal B cells. Additionally, Sézary cells from one patient studied induced a nearly 10-fold increase in IgA synthesis by lymphocytes from a child with ataxia telangiectasia and selective IgA deficiency. Furthermore, these Sézary cells induced more than a 500-fold increase in IgG and IgA synthesis by lymphocytes from a child with Nezelof's syndrome. When Sézary cells were added to normal unfractionated lymphocytes, they did not suppress immunoglobulin biosynthesis. In addition, unlike the situation observed when large numbers of normal T cells were added to purified B cells, there was no depression of immunoglobulin synthesis at very high malignant T-cell to B-cell ratios. These data support the view that Sézary T cells do not express suppressor cell activity. The results presented in this paper suggest that neoplastic lymphocytes from the majority of patients with the Sézary syndrome originate from a subset of T cells programmed exclusively for helper-like interactions with B cells in their production of immunoglobulin molecules.


Antibodies to guinea pig lymphokines. IV. Suppression of the mixed leukocyte culture reaction by anti-lymphokine globulin.

An antiserum, (anti-lymphokine globulin, ALyG) directed against highly purified products of activated lymphocytes, inhibits the proliferation of responding cells in guinea pig mixed leukocyte cultures ((MLC). This serum recognizes three newly synthesized lymphocyte products (one of them being migration inhibition factor (MIF) which are involved in the mediation of delayed hypersensitivity reactions in vivo. Since ALyG does not appear to contain cytotoxic antibodies against guinea pig lymphocyte antigens and its inhibitory activity cannot be removed by absorption with lymphoid cells it seems likely that the inhibition of MLC reactivity is not mediated by the lysis of stimulator or responder cells and its target is not the lymphocyte per se but possibly some factor elaborated during MLC response. By contrast, antisera with specificity for histocompatibility (H) antigens can inhibit the MLC when the appropriate H antigens are present on the responding and/or stimulating cell population. However, this inhibitory of the antisera can be effectively absorbed with lymphoid cells bearing the appropriate H antigens. The addition of ALyG even 48 hr after the initiation of culture results in a marked inhibition of MLC reactivity. This finding is consistent with the elaboration of a mitogenic factor or signal during the first 48 hr of culture and the delivery of this signal to the responding cell population. Thus, ALyG does not appear to interfere with the synthesis of this factor but must be present after its release in order to block responder cell proliferation. Furthermore, the stimulatory effect of this MLC mitogenic factor, but not of PPD-induced mitogenic factor on "third party" cells can be completely inhibited when the cells are cultured in the presence of ALyG. These findings suggest that the MLC-mitogenic factor in this system is identical to or cross-reactive with one or both of the newly synthesized molecules recognized by the ALyG.


Immunity to Plasmodium Berghei yoelii in mice. I. The course of infection in T cell and B cell deficient mice.

The course of infection with 17X nonlethal Plasmodium berghei yoelii was examined in BALB/c mice which were deficient in either T cells or B cells. Markedly increased parasitemia and mortality were observed in athymic (nude) mice which had been backcrossed on a BALB/c background (T cell deficient) compared to similar mice which had been grafted with neonatal BALB/c thymus, and were also observed in BALB/c mice suppressed from birth with goat antiserum to mouse mu-chain (B cell deficient) compared to age- and sex-matched BALB/c controls. These results establish the requirement for the presence of both T cells and B cells for effective resistance to an intercurrent infection with 17XNL P.b. yoelii in adult BALB/c mice. Mechanisms by which the requirement for both T cells and B cells could be explained were discussed. The model of mu suppression was shown to be a valuable tool for an evaluation of the cellular basis of immunity to an infectious disease.


Mechanism of mastocytoma-mediated suppression of lymphocyte reactivity.

Previously we have shown that mitomycin-treated P-815 (H-2d) cells (P-815m) inhibit the in vitro response of C57BL/6 spleen cells to mitogens and DBA/2 alloantigens. The present data indicate that P-815m cells also inhibit the response of C57BL/6 spleen cells to AKR (H-2k)-stimulating cells and that the inhibition does not appear to be the result of crowding. Subpopulations of spleen cells obtained by density gradient centrifucation or after renoval of glass-adherent cells are all sensitive to the inhibitory effects of P-815m. We also show that P-815m cells appears to operate both in vitro and in vivo. The experiments characterize the suppression of MLC reactivity by cell-free preparations of sonicated P-815 mastocytoma cells. The results suggest that the inhibition is not removed by ultracentrifugation, ultra-violet irradiation, or dialysis of the sonicate. P-815 sonicates subjected to chloroform extraction, heating at 56 degrees C, 0.1 mu filtration, or treatment with anit-minute virus of mice serum are still inhibitory. This inhibition is resistant to RNAase treatment but destroyed by pronase. P-815 sonicate suppresses the response of C57BL/6 spleen cells to both DBA/2 and AKR-stimulating spleen. Inhibition of MLC reactivity is only seen if P-815 sonicate is added within the first 48 hr after the initiation of culture. These results indicate that the inhibition of MLC by sonicates of P-815 cells is due to a nondialyzable protein whose effects are not H-2 restricted and are acting on the early phase of sensitization.


Effect of Vibrio cholerae neuraminidase on the generation of cell-mediated cytotoxicity in vitro.

Vibrio cholerae neuraminidase (VCN))12.5 units/2 X 10(6) cells/ml) continuously present for a standard 5-day MLC will significant (p less than 0.02) increase the cytotoxic activity generated by a given number of responding spleen cells without reducing the specificity. Heat-inactiviated VCN produced no such augmentation. This augmented cytotoxicity could be reproduced by preincubating (1 hr) the responding spleen cells with VCN (25 units/2.5 X 10(6) cells/ml) before addition of stimulating spleen cells. Preincubating the stimulator spleen cells with VCN had no effect. VCN preincubation of target cells or presensitized effector cells produced no augmentation. The addition of soluble VCN to the killing assay also did not increase cytotoxicity. Thus, VCN acts only during the generation of specifically sensitized cytotoxic T cells. When the effect of VCN on MLC reactivity, cell recovery and total cytotoxicity (lytic units/10(6) cells) were compared, it became apparent that VCN increases the proliferation of responder cells after stimulation resulting in both an increased number of cells and also an increase in the proportion of specifically sensitized cytotoxic cells in the culture. VCN treatment of responder cell membrane apparently permits a more ready response to allogenic antigens in culture facilitating both increased proliferation and the increased development of specific cytotoxic killers.


Activation of cytotoxic function in T lymphocytes.

The requirements for activation of cytotoxic function in mouse T lymphocytes were investigated. Initial generation of cytotoxicity in normal lymphocytes was equal in magnitude with either Con A or specific alloantigen, and in either case required DNA synthesis. Cytotoxic function in MLC-primed cells could also be regenerated by Con A, the magnitude and target specificity of the cytotoxicity thus generated being indistinguishable from that recalled by specific alloantigen. Cytotoxicity could also be regenerated by third party-stimulating cells; however, the cytotoxicity evoked by third party cells was always specific only for target cells of the original stimulating cell H-2 genotype. The data presented suggest that there are a number of ways to activate cytotoxicity in effector T cells, and are most consistent with a model for T cell triggering that minimizes a strict informational function of antigen-receptor interactions.


The fluorescein-antifluorescein quenching system as a model for assaying changes in antibody function.

A rapid, simple and inexpensive procedure is presented for utilizing the fluorescein-antifluorescein quenching reaction as a model system to monitor and evaluate changes in antibody function brought about by conjugation and modification procedures. Experiments showing 1) the gradual loss of antigen-binding activity during dinitrophenylation, 2) the partial loss and subsequent recovery of activity during the reversible blocking of amino groups by citraconylation and 3) the quantitative release of bound fluorescein during guanidine denaturation were performed to demonstrate the versatility of the model.


Two restriction and modification systems in Staphylococcus aureus NCTC8325.

The presence of two distinct host specificities in Staphylococcus aureus strain NCTC8325 was revealed by the isolation of restriction- and modification-deficient mutants. The two host specificity systems, designated S1 and S2, are both active on phage 80mualpha but are not additive in their restricting activity. Restriction-deficient, modification-proficient mutants were invariably affected in both restriction systems. The functional relationship between these two systems is discussed.


Peritoneal bypass in the treatment of hydrocephalus: historical review and abdominal complications.

The development of intraperitoneal shunting systems is presented chronologically and a review is made of the wide variety of intraperitoneal complications which have occured to date.


The effect of the local anaesthetic bupivacaine on the muscle spindle of rat.

The effects of a long-lasting local anaesthetic, bupivacaine, in combination with hyaluronidase, on the intrafusal muscle fibres of adult rat muscle spindles have been investigated by light and electron microscopy. Necrotic changes in the axial bundle are present within 4 h of the combined drug injection and are widespread by 2 days. The equatorial nucleation of the spindle is lost owing to the necrosis of the myonuclei. Satellite cells, however, appear to survive these changes, even where the underlying muscle fibres are grossly necrotic. Sensory nerve terminals undergo necrosis during this period and plate-type fusimotor nerve terminals withdraw from the degenerating muscle fibres. Macrophage infiltration and early regeneration of the axial bundle is apparent by the third day. Myoblasts first appear at the periphery of the affected muscle fibres, but decrease in number as regeneration proceeds. By the end of the third week, regeneration of the muscle component is complete and re-innervating fusimotor nerve terminals established. The encapsulated regions of the spindle remain abnormal, without recognizable bags or chains of nuclei, although the muscle fibres differ both in size and ultrastructure. Regenerating sensory axons make contacts with the intrafusal bundle that differ in their configuration and ultrastructure from normal terminals. The production of enucleated spindles is discussed in the light of the morphogenetic influences of the spindle nerve terminals, as well as the possible use of such a model in discovering the function of nuclear bags and nuclear chains.


Analysis of muscle receptor connections by spike-triggered averaging. 1. Spindle primary and tendon organ afferents.

1. The synaptic connections of 44 single identified muscle spindle Ia afferents and of 21 Golgi tendon organ (Ib) afferents from medial gastrocnemius (MG) were studied in 46 cats by the spike-triggered averaging of synaptic noise in 803 motoneurons of various types. 2. The well-known monosynaptic Ia excitatory connections were confirmed and their characteristics examined in 113 cells. The method was used at greater sensitivity than before and revealed that, in addition to the larger EPSPs of the order of 300 muV, there were many below the previously reported lower 17-muV limit. 3. By studying the Ia disynaptic inhibitory pathway with quick stretch-evoked Ia volleys and by spike-triggered averaging (STA), it was shown that the latter method can reveal disynaptic and possibly trisynaptic excitatory connections. This is believed to depend on having continuous activity in the relevant interneurons. 4. Latencies of individual connections showed broad distributions and arguments are advanced for setting working limits to mono- and disynaptic paths for Ia excitation and inhibition. Monosynaptic EPSP latency from cord entry was 0.4-1.1 ms and disynaptic inhibition was 1.2-2.4 ms. It was recognized that the boundaries are not rigid and monosynaptic Ia EPSPs may have latencies up to 1.5 ms. 5. Rise times of disynaptic PSPs were, on average, significantly longer than monosynaptic, but individual disynaptic responses could have values within the monosynaptic range. 6. A small diphasic wave shortly preceding the monosynaptic EPSPs was interpreted as a presynaptic spike. Its timing was consistent with this and, as such, permitted estimates to be made of central conduction time. 7. An early negative wave (latency less than or equal 1.1 ms) of small emplitude was sometimes detected in antagonist motoneurons when triggering from Ia afferents. It was found tha extracellular fields could be detected due to single Ia afferent excitations and efforts were made to see if the early negative wave could be explained by this. In a few cases there was evidence that a very short-latency IPSP might be occuring. This evidence and its implications are discussed with attention to the new factors which have to be considered in using the spike-triggered averaging method at very high sensitivity.  8. Ib effect were di- or trisynaptic. They were excitatory to 18% of synergists and to 28% of antagonists. They were inhibitory to 41% of synergists and to 19% of antagonists. The Ib IPSPs were larger than the EPSPs.


Analysis of muscle receptor connections by spike-triggered averaging. 2. Spindle group II afferents.

1. The spike-triggered averaging (STA) method has been used to study synaptic connections of nine spindle group II afferents from medial gastrocnemius to 151 motoneurons of leg muscles in the cat. 2. EPSPs were found in 40 cells, predominantly of triceps surae with latency from cord entry ranging from 0.3 to 4.2 ms. Those with latency less than or equal to 1.4 ms were deduced to be monosynaptic in confirmation of Kirkwood and Sears (13). Mean amplitude for MG-LGS cells was 30.1 muV and mean rise time 1.0 ms (compared with 65.4 muV and 1.0 ms for monosynaptic Ia EPSPs from the preceding report (26)). It is argued that monosynaptic latency for spindle group II afferents could be as large as 1.65 ms. 3. The occurrence of a presynaptic spike permitted the division of EPSP latencies into central conduction time and synaptic delay components. Sindle group II central conduction times were significantly longer than those of Ia afferents, while there were no differences in the synaptic delays associated with the two afferent types. 4. EPSPs of longer latency were judged to be di- or trisynaptic. They were smaller and had longer rise times than the monosynaptic effects. Evidence is presented to show that short rise time of an individual PSP does not guarantee that it is monosynaptic. Rise times are different only on a population basis. 5. Inhibitory responses were found with latencies and mean rise times appropriate for di- and trisynaptic connections. Their mean amplitude was 4.6 muV. 6. The distribution of EPSPs and IPSPs was generally consistent with their exerting stretch reflex effects similar to that of Ia afferents and inconsistent with the inclusion of them in the grouping known as "flexor reflex afferents."


Kinetic modifications of the acetylcholinesterase and (Ca2+ + Mg2+)-ATPase in rat erythrocytes by cholesterol feeding.

The influence of cholesterol on the membrane-bound acetylcholinesterase and (Ca2+ + Mg2+)-ATPase was studied in erythrocytes of five groups of male rats fed different fat-supplemented diets. Two groups of rats were fed essential fatty acid (EFA) sufficient diets with 5% lard or corn oil as the dietary fat, and two groups were fed EFA-deficient diets: a basic, fat-free diet and the same diet supplemented with 5% hydrogenated beef fat. One additional group of rats was fed a stock diet. The kinetic changes recorded were in the degree of the cooperativity of the inhibition by F- of the acetylcholinesterase and the activation by Ca2+, and by Mg2+ of the (Ca2+ + Mg2+)-ATPase. The kinetic behavior of the enzymes was only modified by cholesterol feeding when they were bound to a membrane with a high fatty acid fluidity (e.g. derived from rats fed the corn oil-supplemented diet). The enzymes from a membrane with a low fatty acid fluidity (e.g. derived from rats fed a lard-supplemented diet) were not altered by cholesterol feeding. The changes were noticeable after 24 hours of cholesterol feeding. It is suggested that the in vivo cholesterol sites are involved in a regulatory mechanism for mammalian membrane-bound enzymes.


Properties and physiological effects of thermal carbon black.

Studies have been carried out to determine the possible physiological effects of contact with carbon black made by the thermal process. Contact was made by ingestion, skin application and/or inhalation. Ingestion and/or skin application do not appear to lead to any changes from the normal. Inhalation exposures do not produce pulmonary function changes (antemortem or postmortem studies) but may lead to moderate to severe "perifocal" emphysema in Rhesus monkeys (not observed in guinea pigs). Inhalation studies also suggest right ventricular septal and to a degree, left ventricular hypertrophy in Rhesus monkeys as an effect. Additional studies are necessary to support these finding and consideration must be given to the variability among exposed subjects as compared to controls.


Potassium release from the rat submaxillary gland in vitro. II. Induction by parasympathomimetic secretagogues.

The kinetics of K+ release from an in vitro system of rat submaxillary gland slices were studied after stimulation with parasympathomimetic secretagogues. The slices were incubated at 37degreesC in an oxygenated, enriched Krebs-Ringer bicarbonate medium in the presence and in the absence of Ca++ and of ouabain and, in some experiments, in the presence of the specific antagonists atropine (5 x 10(-6) and 2 x 10(-5) M), phentolamine (2 x 10(-5) M) or propranolol (2 x 10(-5) M. K+ release was elicited by the addition of acetylcholine (2 x 10(-5) M), pilocarpine (2 x 10(-5) M) and carbamylcholine (10(-9) to 2 x 10(-5) M). The results demonstrate that: 1) The selective stimulation of cholinergic receptors induces a rapid net release of K+ from the slices. After 10 minutes of incubation, the percent K+ released after a 2 x 10(-5) M dose of each of the three secretagogues was, respectively, 20.8%, 15.5%, and 19%. 2) The response to carbamylcholine does not occur when Ca++ is absent from the medium and is blocked by atropine but not by phentolamine or by propranolol. Atropine (5 x 10(-6) M) causes a 17-fold shift to the right on the dose-response curve to carbamylcholine. 3) The magnitude of K+ release is the ratio of two opposing mechanisms, a passive efflux and an active reuptake. The latter depends on the activity of the ouabain-sensitive Na+-K+-adenosine triphosphatase. 4) The sensitivity of the slice system to carbamylcholine seems to be greater than that to norepinephrine in terms of net K+ release after equimolar doses of 2 x 10(-5) M and also in terms of the dose required to induce a half maximal passive K+ efflux. However, the maximal passive K+ efflux is similar after both types of secretagogue and amounts of approximately 45% of the K+ present in the slices.


Distribution of fusimotor axons to intrafusal muscle fibres in cat tenuissimus spindles as determined by the glycogen-depletion method.

1. The distribution of fusimotor axons to bag1, bag2 and chain muscle fibres in cat tenuissimus spindles has been studied using a modification of the glycogen-depletion technique of Edstrrom & Kugelberg (1968). Single fusimotor axons were stimulated intermittently at 40-100/sec for long periods (30-90 sec) during blood occlusion. Portions of muscle containing the activated spindles were quick-frozen, fixed in absolute ethanol during freeze-substitution, and then embedded in paraffin wax. Serial transverse sections were stained for glycogen using the periodic acid-Schiff method, and examined for depletion. 2. Dynamic gamma axons (i.e. those that increase the dynamic index of primary-ending responses to ramp stretches of large amplitude) depleted bag1 fibres almost exclusively. 3. Static gamma axons (i.e. those that reduce or abolish the dynamic index) depleted both bag and chain fibres. Bag1 and bag2 fibres were depleted about equally. 4. A single static gamma axon may activate both bag and chain fibres in one spindle (the most common pattern), chain fibres only in another, and bag fibres only in a third spindle. 5. Static gamma axons with conduction velocities less than 25 m/sec also had a non-selective distribution, but no depletion was observed in bag2 fibres. 6. The zones of depletion produced by dynamic gamma axons were distributed more or less equally in the intra- and extracapsular parts of spindle poles, whereas those produced by static gamma axons were mainly intracapsular. 7. The results are compared with the glycogen-depletion studies of Brown & Butler (1973, 1975) and our own study of the distribution of static gamma axons to spindles in which all other motor axons had degenerated (Barker, Emonet-Dénand, Laporte, Proske & Stacey, 1973). The implications of the finding that both static gamma and dynamic gamma axons activate bag1 fibres are discussed.


Laparoscopy in Macaca mulatta: specialized equipment employed and initial observations.

Laparoscopy in rhesus monkeys and specialized equipment developed or adapted for this procedure are described. Repeated laparoscopy in the same animal throughout the menstrual cycle showed by morphological and hormonal criteria that this technique does not significantly influence follicular growth, ovulation, luteal function or cycle length. Observations of the side of ovulation and other follicle/corpus luteum relationships are also described.


Technique of laparoscopy in the chimpanzee.

We have developed the technique of laparoscopy in the chimpanzee using a modification of the standard human procedure. The technique requires careful management, but permits repeated examination, high quality photography, ovarian biopsy, follicle aspiration and injections into the ovary. The chimpanzee may thus be useful for evaluating new techniques or instrumentation intended for human patients. In addition, laparoscopy access to the internal genitalia enhances the value of the chimpanzee as an experimental model.


The development of modern endoscopy.

The development of endoscopic techniques and instruments is traced from Bozzini's light transmitter in the early 1800's to the present. Early use of laparoscopes, cystoscopes, hysteroscopes and culdoscopes is described. While the site of use of these instruments differs, certain general aspects are similar. Modern instrumentation permits visualization into almost any body cavity.


Laparoscopic techniques for biomedical research.

The techniques used for laparoscopic observation of reproductive systems in 23 species are described. The application of various techniques to biomedical research is examined, and the methods used for laparoscopic follicular aspiration and injection, as well as the techniques for uterine flushing collection from swine and nonhuman primates, and oviductal deposition of test solutions are described.


Analytical isolation of plasma membranes of intestinal epithelial cells: identification of Na, K-ATPase rich membranes and the distribution of enzyme activities.

A procedure was developed for the analytical isolation of brush border and basal lateral plasma membranes of intestinal epithelial cells. Brush border fragments were collected by low speed centrifugation, disrupted in hypertonic sorbitol, and subjected to density gradient centrifugation for separation of plasma membranes from nuclei and core material. Sucrase specific activity in the purified brush border plasma membranes was increased fortyfold with respect to the initial homogenate. Basal lateral membrane were harvested from the low speed supernatant and resolved from other subcellular components by equilibrium density gradient centrifugation. Recovery of Na, K-ATPase activity was 94%, and 61% of the recovered activity was present in a single symmetrical peak. The specific activity of Na, K-ATPase was increased twelvefold, and it was purified with respect to sucrase, succinic dehydrogenase, NADPH-cytochrome c reductase, nonspecific esterase, beta-glucuronidase, DNA, and RNA. The observed purification factors are comparable to results reported for other purification procedures, and the yield of Na, K-ATPase is greater by a factor of two than those reported for other procedures which produce no net increase in the Na, K-ATPase activity. Na, K-ATPase rich membranes are shown to originate from the basal lateral plasma membranes by the patterns of labeling that were produced when either isolated cells or everted gut sacs were incubated with the slowly permeating reagent 35S-p-(diazonium)-benzenesulfonic acid. In the former case subsequently purified Na, K-ATPase rich and sucrase rich membranes are labeled to the same extent, while in the latter there is a tenfold excess of label in the sucrase rich membranes. The plasma membrane fractions were in both cases more heavily labeled than intracellular protein. Alkaline phosphatase and calcium-stimulated ATPase were present at comparable levels on the two aspects of the epithelial cell plasma membrane, and 25% of the acid phosphatase activity was present on the basal lateral membrane, while it was absent from the brush border membrane. Less than 6% of the total Na, K-ATPase was present in brush border membranes.


Localization of Na/K-ATPase sites in the secretory and reabsorptive epithelia of perfused eccrine sweat glands: a question to the role of the enzyme in secretion.

Na/K-ATPase sites in both the secretory and reabsorptive epithelia of isolated and microperfused human eccrine sweat glands are localized cytologically. Localization was accomplished through autoradiography of bound 3H-ouabain, a specific inhibitor of the enzyme. Ouabain binding characteristics were determined to ensure maximum specific binding. Enzyme sites are localized only on the basolateral surface of both epithelia in spite of the fact that sodium transport is reversed, i.e., secretory: blood to lumen and reabsorptive: lumen to blood. In view of these findings and in comparison with other recent observations, the role of Na/K-ATPase in secretory electrolyte transport is questioned.


Ruthenium red-staining coat of paired neoplastic and nonneoplastic mouse cell lines in cluture.

The ruthenium red-staining coat (RRSC) of a pair of neoplastic and nonneoplastic cell lines derived from a common pool of mouse embryo cells was examined by electron microscopy. The object was to establish whether a thickening of RRSC is associated with "spontaneous" neoplastic transformation of cells in culture. Ruthenium red has an affinity for acid mucopolysaccharides of the cell coat. Differences in thickness of RRSC between cells of these neoplastic and nonneoplastic lines were negligible.


Evidence for a colon chalone.

The molecular weight fractions of 10,000-50,000 daltons prepared from "used" medium obtained during cultivation of human colon carcinoma cells (SW-48) in vitro inhibited the proliferation and DNA synthesis of these cells. Fractions exceeding 50,000 daltons were not inhibitory; those less than 10,000 daltons were cytotoxic. The inhibitory fraction did not affect either proliferation of human fibroblasts or transformation of human lymphocytes in vitro. Similar fractions from the colon mucosa of other species inhibited the proliferation of SW-48 cells, whereas extracts of dog jejunum or lung did not. This mitotic inhibition was completely reversible and could be destroyed by preincubation with trypsin. Therefore, colon cells appear to contain a cell- (but not species) specific, endogenous mitotic inhibitor or chalone.


Clindamycin and diarrhea.

Of 298 outpatients receiving clindamycin for a variety of indications, diarrhea developed in ten (3.4%); none were diagnosed as having pseudo-membranous colitis. This is well within the range of relative frequencies of diarrhea development (0% to 22.2%) reported in 45 previous studies. Reliance solely on computer-recorded instances of diarrhea yielded a substantial underestimate of diarrhea occurence, but the rate was still significantly higher (P less than .05) in clindamycin users than in nonusers.


Unsuspected substantial pericardial effusions detected by echocardiography.

In a consecutive series of 1,225 routine echocardiograms, a pericardial effusion (PE) was present in 15% and was considered substantial in 10%. Sixty-one percent (68/111) of the latter were clinically unsuspected. A retrospective analysis of these patients showed that 60% (41/68) had underlying heart disease such as congestive heart failure, left ventricular hypertrophy, or asymmetric septal hypertrophy without other discernible causes for PE. In patients referred specifically for suspected pericardial effusion, 67% (29/43) had diseases recognized as causes of pericardial effusion. Unsuspected substantial PE are commonly detected by echocardiography, especially in patients with underlying heart disease.


Peritoneoscopy in Hodgkin disease. Confirmation of results by laparotomy.

Peritoneoscopy was used to evaluate the liver in 35 previously untreated patients with Hodgkin disease. Four were found to have hepatic involvement. Of the 31 patients with normal peritoneoscopies, only one had liver disease demonstrated subsequently at confirmatory exploratory laparotomy. The diagnostic accuracy of peritoneoscopy was 93% for patients at high risk for hepatic disease and 97% for all patients studied. Morbidity from the procedure was minimal. There was no mortality. Peritoneoscopy is a highly accurate staging procedure that should be considered as an antecedent or as an alternative to laparotomy in patients with Hodgkin disease. Its accuracy and minimal morbidity should be considered in the critical selection process of determining which individuals should undergo laparotomy.


Ultrastructural changes in hypertrophied myocardium of spontaneously hypertensive rats.

In seven arbitrarily selected groups of 3-, 5-, 9-, 11-, 15-, and 21-week and one-year-old spontaneously hypertensive rats (SHR) (F25-26), qualitative as well as quantitative light and electron microscope studies were conducted on the myocardium in the left ventricular papillary muscle and lateral wall. Age-and body-weight-matched groups of normotensive Wistar-Kyoto rats were employed as controls. In the strain of SHR used in this study blood pressure was significantly higher than in the controls in the 7th week of age, reached sustained levels (approximately 180 mmHg) in the 15th week of age and remained higher thereafter. The heart weight/body weight ratio gradually increased in SHR after the 15th week of age. In 3-week-old SHR, there was no significant difference in fine structure of the myocardium in comparison with control animals. The cardiocytes were small in size and rapidly growing. Myofibrillogenesis seemed to occur in association with juvenile forms of intercalated discs, subsarcolemmal dense mats, occasional Z band expansions and unorganized filamentous structures in the periphery of preexisting myofibrils. In 5- and 9-week-old SHR, the muscle fiber diameters in the papillary muscle were significantly larger than those in control animals of the same age. Subcellular structures of many cardiocytes were, however, apparently equal in both papillary muscle and lateral wall of the left ventricle in SHR and control animals. In 11-week-old SHR, the cardiocytes were moderately hypertrophied in both papillary muscle and lateral wall. The myofibrils were significantly increased in mass in association with altered intercalated discs, extended subsarcolemmal dense mats and unorganized filamentous structures in the periphery of preexisting myofibrils. Focal interstitial fibroses occurred in in the myocardium, and tunnel capillaries appeared in some hypertrophied cardiocytes. In 15-week-old SHR, the cardiocytes had markedly hypertrophied in accordance with the establishment of sustained hypertension. The intracellular volume ratios of myofibrils and T system had increased significantly whereas the mitochondrial volume had markedly decreased resulting in a significant increase in the myofibril/mitochondria volume ratio. There were marked changes in the intercalated discs associated with numerous immature sarcomers, and frequent abnormal Z band expansions. Interstitial fibroses had markedly increased in number and extent. In 21-week-old SHR, in addition to findings similar to those in 15-week-old SHR, heterogeneous changes of the contractile material, mitochondria and sarcoplasmic matrix were more prominent, suggesting heterogeneous malfunctions in later stages of cardiac hypertrophy. In one-year-old SHR, subcellular changes in many cardiocytes qualitatively resembled those in 21-week-old SHR. Interstitial fibroses and tunnel capillaries were also increased in extent and number...


Biochemical aspects of experimental cardiac hypertrophy.

In experimental cardiac hypertrophy induced by aortic constriction of rats, the hypertrophy was established after 5-7 days. The basic biochemical changes for increasing tissue mass; increases in protein, nucleic acid, and polyamine synthesis started to occur between 2 and 8 hours followed by an increase in uridine nucleotide pools via predominance of "salvage" pathway. Although the precise coupling mechanism between mechanical stress and biochemical changes is still obscure, an interval between increased load and DNA transcription may be quite short. Some of the key enzymes regulating these processes showed a biphasic response the reason of which is not clear. The established hypertrophied heart muscle showed a decrease in velocity of isotonic shortening and an increase in resting tension. The former alteration is referred to a decrease in myosin ATPase activity and an disorder in excitation-contraction coupling mechanism, and the latter is supposed to be due to an increase in collagen in heart muscle.


Hemodynamic studies on hypertrophied hearts in man.

A total of 31 patients were studied during cardiac catheterization. Hemodynamics in 9 cases with cardiac hypertrophy secondary to chronic volume load (MI, AI) and in 9 cases with cardiomyopathy were compared to those in 5 normal cases, 5 cases with MS and 5 cases with near normal left ventricle. A hypertrophied left ventricle had an enlarged cavity, an increased wall mass and an elevated systolic midwall tension. Peak systolic stress in left ventricular wall was normal in compensated patients with volume load, but below normal in compensated patients with cardiomyopathy. Peak systolic stress was elevated in decompensated patients either with regurgitation or with cardiomyopathy. The significance of this stress difference between volume over load and cardiomyopathy was discussed. Depressed pump function was observed in hypertrophied left ventricle especially in cardiomyopathy.


Outcomes of children with rheumatic fever not diagnosed by revised (1965) Jones criteria.

The Jones criteria proposed in 1944 for the diagnosis of rheumatic fever (RF) underwent a modification in 1955, and then a revision in 1965. The importance of establishing antecedent streptococcal infection was stressed, and the criteria became more difficult to meet with. Thirty-two children, whose clinical and laboratory manifestations met the modified but not the revised Jones criteria were encountered at the National Taiwan University Hospital during 1967-1971. They were labeled as probable RF, and were treated and follwed for 1 month to 6 years (average 3.7 years). The diagnosis in each case was evaluated in the light of their outcomes. The diagnosis of RF was justified in 27 cases based upon the following observation: Favorable responses to anti-rheumatic treatment (27 cases); normalization of the enlarged heart with disappearance of the murmurs (3 cases); normalization of the enlarged heart with persistence of the murmurs (5 cases); significant reduction of the enlarged heart (12 cases); typical RF recurrence (8 cases); development of pure mitral stenosis (1 case); valvular pathology verified at surgery or autopsy (4 cases). The diagnosis in the rest of 5 cases remained not confirmed or negated because that: the enlarged heart stayed unchanged (1 case); the child remained uneventful and free of cardiac involvement (2 cases); and the patient died and no postmortem study obtained (2 cases). The risk factors leading to underdiagnosis in these patients are: 1) late coming under observation; 2) no response in ASO titers; 3) limited studies for the evidence of streptococcal infection; and 4) drug induced modification of the clinical manifestations. This study implicates that patients who are very suspicious of RF on the clinical grounds, yet fail to meet the revised Jones criteria, especially those with established valvular heart disease, should be labeled as cases of probable RF, and be treated, followed, and placed on prolonged chemoprophylaxis until proved otherwise. Recurrence of RF may thus by prevented, and the regression or even natural healing of the rheumatic heart disease will become a possibility.


Giant T wave inversion as a manifestation of asymmetrical apical hypertrophy (AAH) of the left ventricle. Echocardiographic and ultrasono-cardiotomographic study.

Left ventricular scanning by echocardiography and ultrasono-cardiotomography was performed to search the possible muscular abnormality in 9 cases with giant T wave inversion without documented cause. The deeply inverted T wave was more than 1.2 mV (average was 1.63 mV) in the left precordial leads. All the cases had electrocardiographic left ventricular hypertrophy of obscure origin and ischemic episode was absent. Conventional echo beam direction to measure the short axis of the left ventricle disclosed almost normal thickness and movement of both interventricular septum (IVS) and the posterior wasll (PW), so that the report of these cases is frequently within normal limits. However, ultrasono-cardiotomography (sector B scan) disclosed the fairly localized hypertrophy near the left ventricular apex, and conventional echocardiography also revealed the same area of either IVS or PW or both below the insertion of the papillary muscles, when the scanning towards the apex was performed (asymmetrical apical hypertrophy: AAH). Control study of 9 cases with IHSS showed asymmetrical septal hypertrophy (ASH) with almost equally hypertrophied IVS from base to apex. All cases had inverted T waves, but these were of lesser degree. Three cases had relatively deep T wave compatible with those of AAH, and these cases also had the apical hypertrophy of considerable degree (unusual type of IHSS, i.e., intermediate type between AAH and ASH). The close relationship between the depth of the inverted T waves and the Apex/Mid wall thickness ratios suggests that the altered recovery process of the hypertrophied apical musculature is responsible for the giant T wave inversion of heretofore unsolved origin. Until the connective link of AAH to the other forms of hypertrophic cardiomyopathy is disclosed, the cases with such a T wave and the apical hypertrophy may be designated as asymmetrical apical hypertrophy (AAH).


The neuropathology of subacute myelo-optico-neuropathy, "SMON", in the humans: --with special reference to the quinoform intoxication.

The detailed neuropathology of the 16 autopsy cases of the acute to chronic "SMON" of different ages together with those reported before and at the "SMON" Research Commission has been described and reviewed. This was compared with the alreadyknown neurological disorders, particularly systemic and/or pseudosystemic degeneration of the long tracts of the spinal cord with polyneuropathy together with the "SMON"-like diseases caused by certain neurotoxic and/or neurotropic agents. It has been concluded that "SMON" can be belonged to a subgroup of some endemic diseases within the main group of combined degenerations associated with complex nutritional deficiency or toxic interference of tissue metabolism. Since an excess and mal-administration of quinoform in quantity, time and space were identified in a great majority of the autopsy cases, quinoform could comprize the most important etiology for the "SMON" onset. This view also was supported by the almost complete reconstruction of the neuropathology of the human "SMON" in the experimental animals intoxicated with quinoform which disclosed a precise neurotropism from the autoradiograms of the same animals. As compared with the similar lesions in SLE,hepatocerebral diseases, etc., the pathogenesis and clinicopathological relationship of "SMON" was speculated. The accerelating and/or modifying factors for the SMON" onset aside from quinoform were discussed.


Neurotoxity of iodoxyquinoline: a further study on beagle dogs.

Eight beagle dogs were treated orally with iodoxyquinoline (chinoform, clioquinol) again, and all of them manifested similar neurologic symptoms to those observed in our previous experiments on mongrel dogs, beagle dogs, cats and a monkey. The neurologic symptoms were identical both clinically and pathologically with those of subacute myelo-optico-neuropathy (abbreviated to SMON) in man. In the present study, the neurologic manifestations evolved with less total chinoform doses (5.1-13.8 g/kg) in shorter periods (19-47 days) and there was no difference between each group on the increasing dosage and on fixed dosage. It is discussed in the present paper that onset of the neurologic symptoms is associated with unconjugated chinoform levels in the blood.


Prognosis of SMON patients.

The following points have become clear on prognosis of SMON through the analysis of 981 cases collected. 1) The prognosis of the old whose ages were 60 year old or over is not favorable, when compared with that of the young. However, there is no prognostic difference between male and female. 2) The cummulative death rate of SMON which was calculated by the life table method is approx twice as much as the generally expected value. 3) Approximately 80% of the patients showed some sort of improvement 7 to 12 months after the onset of the disease. The rate for 13 months or over if nearly the same. 4) The abdominal symptoms found at the time of the onset of the disease decreased markedly in the course of the disease. 5) Among neurological symptoms, the prognosis of motor disorders is more favorable. The complete recovery of sensory disturbances was extremely rare, but approx 60% showed more or less favorable in the course of the illness. Approximately 40% of the cases with visual disturbances completely recovered or showed favorable improvement, whereas 9% of them became worse. As for the prognosis of visual impairment, it is more serious than other symptoms. 6) The patients who had been administered clioquinol over long period displayed a higher rate of severe or moderate motor, sensory and visual disturbances, compared with the group with short-term administration of clioquinol. The death rate was also higher in the former group. 7) The rate of relapse as a whole was 16.7% and 68% of them was seen within 18 months after the onset. There is no difference in relapse according to sex. There was seen a high rate of relapse in the group of longterm administration of clioquinol. 8) A 10.5% of total cases were either unable to walk or in need of assistance in walking, whereas the rate of patients who cannot get dressed or who cannot defecate unassisted was lower. 9) Approximately 65% returned to the job in 12 months or more after the onset. The employment rate was not different according to sex, whereas it was lower along with the age advances. 10) Approximately 20% were not received medical treatment. The rate of non-treated patients is higher in the younger patients. The rate of hospitalized patients was higher in the older patients.


[Some metabolic values of acid mucopolysaccharides in myopia (author's transl)].

The study group consisted of 82 children aged 9 to 14 years with the following forms of myopia: progressive (37 cases), stable (22 cases), congenital stable (11 cases) and incipient (12 cases). During the period of progression in myopia, a significant increase was found in the 24-hour-excretion of acid mucopolysaccharides. In the patients with stable and congenital stable forms the excretion of acid mucopolysaccharides did not differ significantly from that in the control group of emmetropic and hypermetropic children. The results of electrophoresis on cellulose acetate and of the carbazol and orcinol reactions showed predominant excretion of A and C chondroitinsulphates in patients with myopia.


Morphometry and autoradiography of early hypertrophic changes in the ventricular myocardium of adult rat: an electron microscopic study.

The ultrastructural composition of left ventricular myocardium and myocytes has been investigated at 20 hours after the surgical production of a subdiaphragmatic aortic stenosis in the adult rat. Morphometric analysis shows a 20% increase in the size of myocytes resulting principally from a 36% increase in mitochondrial volume and a 78% increase in the volumes occupied by smooth endoplasmic reticulum and matrix. Myofibrillar volume is increased only 4%. Measurements of the size and shape of individual mitochondria indicate that the augmentation of this compartment is wholly accountable by the enlargement of preexisting mitochondria. Quantitative autoradiographic analysis following a 2-hour pulse labeling with 3H-leucine shows that the rate of incorporation of newly synthesized proteins is practically uniform throughout all structural components of the sarcoplasm 18 to 20 hours after aortic banding. The conclusion is reached that this interval of hypertrophic response is a transition period characterized by an increasing rate of synthesis of contractile proteins overtaking a previously greater rate of mitochondrial synthesis.


Surface alterations induced by stress in gastric mucosa: protective effect of zolimidine. A transmission and scanning electron microscope investigation.

The fine structure of gastric epithelial cells of normal rats and of rats restrained at low temperature has been examined by transmission and scanning electron microscopy. The observation of the normal mucosa has revealed that in comparison with the cells overlying the flat surface of the stomach, those covering the plicae have a different surface structure with numerous microvilli and a peculiar organization of intercellular junctions. Restrained animals initially showed swelling of the foveolae that was followed by a detachment of the superficial cells leading to severe stomach erosions. It is postulated that the first modification induced by stress is a decrease in the amount of mucus granules and a weakness of the mucous protective barrier. This was confirmed by the results obtained by pretreating rats with zolimidine. This substance, which has been shown to increase secretion and synthesis of gastric mucus, completely protected the animals from the stress-induced damage of the stomach surface. Furthermore, this treatment produced an enlargement of the cisternae of the endoplasmic reticulum and an increase in the surface of the Golgi apparatus in the epithelial cells.


Some changes in the properties of dynein ATPase in situ and after extraction following heat treatment of cilia.

Glycerol-extracted cilia from Tetrahymena pyriformis were demembranated by treatment with Triton X-100 and then heated for up to 30 min at temperatures between 34-38 degrees C. Heat treatment caused an uncoupling of the ATPase from motility as indicated by an increase in ATPase activity and a loss of pellet height response. After heat treatment, the ATPase activity of the dynein in situ differed from that in unheated cilia as shown by an increased sensitivity to a lower temperature of assay (0 degrees C) and by a loss of the activation normally observed upon reaction with N-ethylmaleimide or p-phenylenedimaleimide. Upon extraction of the heat-treated cilia by Tris-EDTA, there was a large loss in ATPase activity so that the heat-treated cilia yielded a crude dynein fraction with a lower specific activity compared with that obtained from unheated controls. The difference was not due to a change in the amount of protein recovered or in the amount of ATPase activity which remained unextracted. Resolution of the crude dynein by sucrose density sedimentation indicated that activity was lost from both the 14S and 30S peaks but more so from the latter than from the former. Thus dynein in situ in cilia in which the ATPase has been uncoupled from motility by gentle heat treatment differs in several important respects from dynein inside unheated cilia.


Functional organization of the outer membrane of escherichia coli: phage and colicin receptors as components of iron uptake systems.

The functional interaction of outer membrane proteins of E. coli can be studied using phage and colicin receptors which are essential components of penetration systems. The uptake of ferric iron in the form of the ferrichrome complex requires the ton A and ton B functions in the outer membrane of E. coli. The ton A gene product is the receptor protein for phage T5 and is required together with the ton B function by the phages T1 and ø80 to infect cells and by colicin M and the antibiotic albomycin, a structural analogue of ferrichrome, to kill cells. The ton B function is necessary for the uptake of ferric iron complexed by citrate. Iron complexed by enterochelin is only transported in the presence of the ton B and feu functions. Cells which have lost the feu function are resistant to the colicins B, I or V while ton B mutants are resistant to all 3 colicins. The interaction of the ton A, ton B, and feu functions apparently permits quite different "substrates" to overcome the permeability barrier of the outer membrane. It was shown for ferrichrome dependent iron uptake that the complexing agent was not altered and could be used repeatedly. Only very low amounts of 3H-labeled ferrichrome were found in the cell. It is possible that the iron is mobilized in the membrane and that desferri-ferrichrome is released into the medium without having entered the cytoplasm. Growth on ferrichrome as the sole iron source was used to select revertants of T5 resistant ton A mutants. All revertants exhibited wild-type properties with the exception of partial revertants. In these 4 strains, as in the ton A mutants, the ton A protein was not detectable by SDS polyacrylamide gel electrophoreses of outer membranes. Albomycin resistant mutants were selected and shown to fall into 5 categories: 1) ton A; 2) ton B mutants; 3) mutants with no iron transport defects and normal ton A/ton B functions, which might be target site mutants; 4) mutants which were deficient in ferrichrome-mediated iron uptake but had normal ton A/ton B functions. We tentatively consider that the defect might be located in the active transport system of the cytoplasmic membrane; 5) a variety of mutants with the following general properties: most of them were resistant to colicin M, transported iron poorly, and, like ton B mutants, contained additional proteins in the outer membrane. The outer membrane protein patterns of wild-type and ton B mutant strains were compared by slab gel electrophoresis in an attempt to identify a ton B protein. It was observed that under most growth conditions, ton B mutants overproduced 3 proteins of molecular weights 74,000-83,000. In extracted, iron-deficient medium, both the wild-type and ton B mutant strains had similar large amounts of these proteins in their outer membranes. The appearance of these proteins was suppressed by excess iron in both wild-type and mutant. From this evidence it is apparent that the proteins appear as a response to low intracellular iron rather than being controlled by the ton B gene...


The closure of abdominal wounds in cancer patients.

A total of 102 abdominal surgical wounds in cancer patients were closed with absorbable suture material. The object of the study was to evaluate whether patients with cancer having possible wound healing impairments could be closed with absorbable sutures, thereby omitting the difficulties involved with retention sutures or nonabsorbable material. Polyglycolic acid sutures were used in the fascia in all of these patients, and they were studied regarding the incidence of wound infection, wound dehiscence, and incisional hernias. There were no instances of wound dehiscence in the entire series. A wound infection rate of 14.8% was encountered. The incidence of incisional hernia following either infection or primary healing was noted to be markedly decreased. The rate of wound dehiscence and wound hernia was sufficiently low to lead us to recommend this type of abdominal wound closure in all cancer patients.


[The fistula of the first branchial cleft A critical presentation of its pathogenesis and its clinical and surgical problems. A description of one case combined with a fistula auris congenita (author's transl)].

A case of congenital fistula of the first branchial cleft (Ohr-Hals-Fistel) is presented. Based on the previously published cases the anatomical and clinical aspects of the disease are summed up. Pathogenesis is discussed on the underlying principles of embryological basic research, centering on Hochstetter's "Entwicklungsgeschichte der Ohrmuschel und des äusseren Gehörganges des Menschen" (1948), a publication that is hardly known. Some hypotheses on the genesis of the fistula of the first branchial cleft are criticised and a new hypothesis is developed to the effect that the fistula is caused by a disturbance in the development of the external ear channel. A few critical remarks are added concerning the discussion on the genesis of fistula auris congenita.


[Animal experiments concerning the neurotoxicity of aminoglycosid antibiotics. Electron microscopic findings regarding dose--depending mitochondrial damage in the cochlear nucleus of the guinea pig (author's transl)].

In 34 guinea pigs (plus 8 healthy animals for control) Gentamicin, Tobramycin and Amikacin show a characteristic pattern of mitochondrial damage in the cochlear nucleus of the guinea pig after a 10-day-administration of 4/40 mg/kg body weight Gentamicin, 4/50 mg/kg Tobramycin, respectively 15/75/150 mg/kg Amikacin. Amikacin causes the most pronounced physiological damage observed by the early loss of Preyer reflex and general toxic signs--these observations correspond with the morphological findings. By the evaluation of morphological findings we are convinced that there is indication for a direct neurotoxic effect on mitochondria in the cochlea nucleus regarding the dose and time of administration.


[Morphological findings of the mucosa after irradiation with an argon-laser an animal experiment (author's transl)].

The macroscopical and lightmicroscopical findings at the mucosa of the hamster cheek pouch after irradiation with an Argon-Ionlaser (wave-length 0.488 mum, power 200 mW, beam diameter 0.5 mm, power density 100 W/cm2) are described immediately, 1; 2,5; 5 and 10 days after irradiation. The mucosa was exposed 10 sec, 30 sec, 1, 2, 5 and 10 min to the radiation. With growing laser dosis the lesions of the mucosa and the postoperative healing time increases.A clinically useful optimal mucosa lesion with a short healing duration (within 5-10 days) is obtained only by application of a low and a middle laser dosis. There are two kinds of mucosa destruction by laser: 1. The destruction can be producing a coagulation necrosis with subsequent unbloody sloughing, resulting in an ulcus with rapid healing by complete reepithelisation and a small amount of connective tissue. 2. An immediate destruction is obtained by carbonisation of tissue after application of a middle to high laser dosis resulting in an ulcus with slow healing and also complete reepithelisation and small connective tissue. The typical laser lesion consists of 3 zones: The carbonisation zone is surrounded by a small coagulation zone followed by a zone of granulation tissue including thrombs and necrosis of the vessel walls. The unbloody procedure is based on an occlusion of the vessels immediately during laser radiation and later on by thrombosis in the periphery of the lesions. The possible clinical applications are discussed.


[A study on the closure of radical cavities. Animal experiments (author's transl)].

Preserved homograft bone chips were implanted into the middle ears of 12 rabbits with close contact of the graft with the bony wall of the bulla. Bone new formation was observed in all grafts, but remodelling was incomplete after a year. The bone chips were connected to the graft bed by new host bone. The homograft was identified by radioactive labelling. For bone new formation the intercellular substance of the transplant was removed by thost tissue and was partly reused in bone new formation.


[Penicillin levels in tonsillar tissue and serum after intramuscular and oral application (author's transl)].

Penicillin levels were determined in both serum and tonsillar tissue 1-2 h (29 patients) and 3-4 h (16 patients) after intramuscular application of Penicillin-G resp. oral doses of Penicillin-V. All tonsils were found to contain different amounts of Penicillin. Concentrations of antibiotic in the tonsillar tissues are dependent on the respective serum levels. The serum-tonsil-difference of the Penicillin levels increases significantly in adults. The efficiency of oral and intramuscular Penicillin application and the occasionally described failure of oral Penicillin medication are discussed with regard to the minimal inhibitory concentrations (MIC) for group A streptococci.


[Classification of malignant lymphomas of the neck: combined morphological, immunocytological, serological and tissues culture studies (author's transl)].

81 untreated malignant lymphomas of the neck were classified morphologically according to the German Kiel classification and to the American classification of Rappaport and Berard and these tumors were typed immunologically as to their T- or B-cell nature. Cells from 16 of these patients were subsequently grown in tissue culture for periods up to seven months. Tissue culture cells were monitored as to spontaneous variations in the morphologic cell type and to the expression of T- or B-cell surface determinants. In addition in 10 patients sera were tested for anti-Epstein-Barr virus (EBV) antibodies. The results of these investigations were correlated with the course of the individual neoplastic disease. Significantly elevated titers against EBV antigens were detected primarily in 8 of 10 patients, mainly in lymphocytic lymphomas respective lymphoplasmacytoid immunocytomas. All such neoplasms belonged immunologically to B-cell lymphomas and were readily grown in tissue culture. The morphological cell type and the expression of B-cell determinants showed some variation during the culture period. In contrast,lymphomas of EBV-negative patients or patients with low EBV-titers grew poorly in tissue culture and remained morphologically more stabile. Immunocytologically they belonged to tumors with B- and T-cell deficiency and were classified primarily as histiocytic lymphomas and as Hodgkin's lymphomas. The clinical course in slow proliferating tumors seemed to be rather disadvantageous.


[Simultaneous estimations of flowrate, protein, immunoglobulin a and lysozyme secretion of human parotid glands with whartin tumours and scintigraphic and sialographic examinations (author's transl)].

7 patients with uni- and bilateral Whartin tumors are examined. In comparison to sialographic and scintigraphic findings simultaneous estimations of flowrate, total protein, immunoglobulin A and lysozyme excretion of these glands in rest and under stimulation are done. Thereby the following results were found: 1. Flowrate is normal like in glands with pleomorphic adenomas. 2. Total protein secretion is significant lower in stimulated secretions. 3. Immunoglobulin A secretion is diminished in stimulated secretions, lysozyme is diminished under rest and stimulation.


[Reconstructive surgery of the lips (author's transl)].

In reconstructive surgery of the lips after resection of tumours the method of Fries will grow to be a most universal procedure. Clinical examples show the advantage of this method which, especially in case of necessary excision of the regional lymphnodes can successfully be combined with a methodical variation.


[Excessive auricular keloid after correction of a protruding ear (author's transl)].

Report of a large auricular keloid after a correction of a protruding ear. Healing by excision and dermatoplasty.


[The prognostic evaluation of Bell's palsy by combined functional diagnosis (author's transl)].

In 82 patients with Bell's palsy nerve excitability test (NET) and electrogustometry (EGM) were performed and the stapedius muscle reflex (SMR) was measured on the day of admission. The comparison of test results with the final outcome of the disease allowed the following observations: in cases with normal SMR a soon and complete regeneration may most probably be expected. The EGM seems to be of prognostic value during the first couple of days after onset only; together with a loss of stapedial function a threshold difference of 15 EGU or more indicates an unfortunate course of the disease. The NET does not say anything about prognosis before the third day after onset. When the threshold of electrical excitability on both sides differs 3.5 mA or more an incomplete recovery has strongly to be expected; however, during the first two weeks a normal excitability does not exclude an incomplete recovery; only after that time such finding means a good prognosis. Those cases with pathological results in all three function tests made a--sometimes severe--defective recovery. These observations suggest, that the above function tests supplement each other very well and that a combined application of these tests can improve prognostic evaluation of Bell's palsy. It seems of special value that by means of these tests cases with a bad prognosis may be already recognized on the third or fourth day after onset with a relatively high reliability.


[Neurootological diagnostic in children by means of cranio-corpo-graphy (author's transl)].

Among the different tests of the neurootological test battery, especially Cranio-Corpo-Graphy, is very useful for objectively investigating children. This method can be performed together with the Unterberger's stepping procedure. Then the blind-folded patient has to make 80-100 steps on the spot. This procedure should be completed within 1 min. For the fotographic recordings lights are installed in a helmet which is placed on the head and are also attached to both shoulders. The Polaroid-film is exposed during the whole duration of the test. Thus the body and head movements can be recorded as light tracings. For evaluating the tracings quantitatively it is necessary to superimpose in a second exposure a coordinate system. We usually take a polar coordinate system which is been produced in the hite of the head of the patient. Typical reactional patterns are used for differentiating peripheral from central or combined peripheral and central vestibular spinal lesions. Especially the Unterberger's stepping procedure is apt to be applied on the children's gaming behaviour. In cases of congenital or early children's torticollis, the amount of angular deflection of the head can be measured by means of Cranio-Corpo-Graphy.