Histochemical investigations on the localization of the purple acid phosphatase in the bovine spleen

Summary The localization of the purple tartrate-resistant, iron-containing acid phosphatase in the bovine spleen was studied by enzyme histochemistry at the light and electron microscopic levels as well as by immunohistochemistry. The purple phosphatase was localized only in lysosome-like organelles of cells belonging to the reticulo-phagocytic system. The same cells were identified as containing large iron(III)-deposits as ferritin in homogeneously granular accumulations and freely in the cytoplasm, or as hemosiderin in siderosomes. The phagocytosing cells containing purple phosphatase and ferritin often had close contact with clusters of aged and deformed erythrocytes.A possible catabolic role of the purple enzyme as a phosphatase degrading phosphoproteins of the erythrocyte membrane and the cytoskeleton was assumed.     

Histochemical investigations on the localization of the purple acid phosphatase in the bovine spleen

The localization of the purple tartrate-resistant, iron-containing acid phosphatase in the bovine spleen was studied by enzyme histochemistry at the light and electron microscopic levels as well as by immunohistochemistry. The purple phosphatase was localized only in lysosome-like-organelles of cells belonging to the reticulo-phagocytic system. The same cells were identified as containing large iron(III)-deposits as ferritin in homogeneously granular accumulations and freely in the cytoplasm, or as hemosiderin in siderosomes. The phagocytosing cells containing purple phosphatase and ferritin often had close contact with clusters of aged and deformed erythrocytes. A possible catabolic role of the purple enzyme as a phosphatase degrading phosphoproteins of the erythrocyte membrane and the cytoskeleton was assumed.     

Improvement of the micromethod for the limulus lysate test.

Frauch's micro-slide method was improved to facilitate the endpoint-determination of the Limulus test. Two precise observations, by inverted phase contrast microscopy and with a staining procedure, were newly performed as additions to the slide test. The staining procedure was proposed as an improved method for the Limulus test since it is simple and convenient. In the staining method, bromophenol blue (BPB) solution was used as the staining solution. A negative (-), a strong positive (++) and a weak positive reaction (+) were characterized by a "ring" formation, a "cloud-like" spread of gel and a "spot" in the "cloud" respectively. Since the distinction between (-) and (+) reactions was obvious in the proposed method, determination of the endpoint was easier than in the ordinary tube and Frauch's method. The sensitivity of the present method was equal to or higher than that of other methods. Inverted phase contrast microscopy was utilized to confirm the findings obtained by the staining method. The volume of the lysate used in this method was as little as 1/10 of that used in the tube method.     

Anatomical variations of the extensor tendons to the fingers over the dorsum of the hand: a study of 50 hands and a review of the literature

The extensor tendons to the fingers were studied in dissections of 50 fresh cadaveric hands, and the divisions of the tendons, as well as the communications (juncturae), were analyzed. The pattern of distribution most frequently observed was as follows. The extensor digitorum communis provided one tendon to the index finger, one to the middle finger, two to the ring finger, and none to the little finger. The extensor indicis exhibited one tendon, whereas the extensor digiti minimi exhibited two tendons. The extensor indicis tendon was always observed to lack a junctura tendinum. The extensor indicis was absent in both hands of one cadaver. A tendon slip from the extensor digiti minimi to the ring finger was observed in one hand. All surgeons must bear in mind the existence of these variations when performing common tendon transfers.     

Immobilization of the exo-maltohexaohydrolase by the irradiation method

Exomaltohexaohydrolase (E.C.3.2.1.98) was immobilized by radiocopolymerization of some synthetic monomers which were mixed in various combinations. Irradiation was carried out while the mixture of monomers and enzymes was frozen in petroleum ether-dry-ice bath. Recovery of the immobilized enzyme was 44-75%.The optimum pH of the enzyme slightly shifted to the acidic side. The pH stability was improved remarkably by immobilization. The enzyme was stable retaining more than 90% of its original activity in the range pH 4-11. The optimum reaction temperature of the enzyme increased about 2 degrees C. Heat stability was also improved by immobilization, and that the enzyme retained about 40% of its original activity after treatment at 75 degrees C for 15 min. The immobilized enzyme was stable to the repeated use of 20 cycles. The K(m) value of the enzyme for short-chain amylose was almost the same as that of native enzyme. When soluble starch was used as the substrate, the K(m), value of the enzyme was three times as large as that of native enzyme. Effects of various metal ions and inhibitors on the immobilized enzyme were also studied compared to the native enzyme.     

From the saltpan to the plate: An evaluation of the use of the edible halophyte Salicornia ramosissima in catering

The halophyte species Salicornia ramosissima has long been consumed as food and is currently regarded as a high-value gourmet item. However, given that these plants grow in estuarine areas, often impacted by pollution, and are prone to the accumulation of ions, the risks associated with food borne pathogens or toxic metals has to be carefully considered. The objective of this work was to gather evidence that S. ramossissima harvested locally in an estuarine system (Ria de Aveiro, Portugal) can be safely consumed as a healthy and catering-amenable food. Fresh shoots, taken as green vegetables, and dried powdered shoots (green salt) intended as seasoning, were analysed for total aerobic bacteria, total coliforms, Escherichia coli, Bacillus cereus and fungi (yeasts and moulds). The nutritional profile and the concentration of macro and micro elements, including potential toxic elements, was also established. Fresh shoots and green salt were incorporated in catering meals (soup and a meat dish) and subjected to sensory analyses. The microbiological quality of fresh shoots and green salt was compliant with the guidelines for salad greens and herbs and spices, respectively. The nutritional and elemental profile revealed that fresh shoots have a high content in fibre and NaCl. From the concentration of Na determined in green salt, a daily dose of 7 g was estimated as corresponding to the recommended daily dietary intake Na. None of the other elements was present in concentrations that could exceed the dietary intake limits. In the sensory tests, fresh shoots were well accepted as salad greens but green salt, used as seasoning, was considered as insufficient in terms of saltiness and does not completely substitute cooking salt. The results confirm the nutritional and sensory value of S. ramossisima and represent a promising prospect for the incorporation of locally harvested or crop cultivated plants in catering meals.     

The role of the ganglioside lipid moiety in the process of ganglioside-cell interactions.

The role of the ceramide moiety of gangliosides, together with the deriving aggregative properties of ganglioside in solution, in the process of ganglioside-cell interactions was studied. The natural GM1(stearoyl) and the synthetic GM1(acetyl), containing the stearoyl and acetyl groups as the acyl moiety, respectively, were used in binding experiments to rat cerebellar granule cells. Regardless of the cell culture conditions, such as the presence of absence of fetal calf serum, the association of GM1(acetyl) to the cells was much greater than that of GM1(stearoyl). GM1(acetyl) was present in the incubation medium as monomers. After incubation, a large part of the total GM1(acetyl) associated to cells, 76-93% depending on the experimental conditions, was removed by washing with protein solutions. The remaining associated ganglioside was not removed by repeating washing with protein solutions or trypsin treatments and was considered as a component of the membrane. The cell association of GM1(stearoyl), present in solution as monomers as well as micelles, could be classified as serum-labile, trypsin-labile and trypsin-stable. The trypsin-stable form of association, corresponding to the molecules stably inserted into the membrane, was proportionally higher, the proportions varying with increasing incubation time and decreasing ganglioside concentration. This form of association was particularly high when incubation was performed in the presence of fetal calf serum. Incubation experiments performed with a mixture of GM1(stearoyl) and GM1(acetyl) in a molar ratio which allowed their presence in the medium as monomers as well as mixed micelles, led to a ganglioside association suggesting that besides the aggregative properties of the molecule other ganglioside properties are involved in the ganglioside-cell interaction process.     

Influence of testosterone on the development of the ischiocavernosus muscle of the rat

The mean area of the neuromuscular endplates and the acetylcholinesterase (AChE) activity at the myotendinous junction of the ischiocavernosus muscle were studied in normal, castrated and testosterone-treated castrated Wistar rats by the Koelle method. The mean endplate area was found to be smaller in castrated rats, compared to normal ones (p less than 0.001), while testosterone treatment restored its normal size (0.8 greater than p greater than 0.7). The terminal AChE activity in castrated rats was as strong and spread as in juvenile ones, while it was almost absent in normal and in testosterone-treated castrated rats. The same parameters were examined in the tibialis anterior muscle of the same rats, chosen as a specimen of 'nonhormone-dependent' muscle, without finding any difference among the single groups.     

Studies on the effect of 2-hydroxy-5-nitrobenzyl bromide on the mechanism of insulin.

The effects of HNB were tested on the binding of insulin to fat cells and fat cell membranes as well as the responsiveness of fat cells to insulin. Although metabolism was found to be inhibited, the reagent was found to stimulate the binding of insulin. The stimulatory effect on insulin binding as well as inhibitory effect on metabolism was reversed by free tryptophan consumption of the reagent. The results are discussed relative to these findings.     

The role of the methylation in the detoxication of arsenate in the rabbit

The biotransformation, tissue retention, intracellular binding and biokinetics of arsenic were studied in rabbits exposed to [74As]arsenate (0.4 mg As/kg body wt., i.v.). Inhibition of the methyltransferase activity by injection of periodate-oxidized adenosine (PAD) caused a marked decrease of the formation of [74As]dimethylarsinic acid (DMA), which gave rise to 1.5-4 times increased tissue levels of 74As. This is almost the same as reported for rabbits given arsenite in combination with PAD and was due to a rapid reduction of the arsenate to arsenite which bound to the tissues. Only about 30% of the arsenate given was excreted unchanged in the urine, indicating that a large part was reduced to AsIII. Thus the methylation to DMA seems to be almost as important for the detoxication following exposure to arsenate as that following exposure to arsenite. In the rabbits with normal methylating capacity 50-70% of the produced AsIII was methylated to DMA. The liver was the only organ in which DMA was present 1 h after the administration, indicating that this is the main site of the methylation. The DMA was rapidly cleared from all tissues except the thyroid.     

Conditions of formation of the heparin-fibronectin-collagen complex and the effect of plasmin

The formation and composition of the insoluble heparin-fibronectin-collagen complex and its degradation by proteolysis was investigated. At fixed concentrations of the other molecular components of the complex, the maximal rate of complex formation, measured turbidimetrically, was reached at a concentration of 4 microM heparin and 0.9 microM collagen, while the rate of complex formation was linearly related to concentrations of fibronectin as high as 3 microM. Heparin was incorporated into the complex in a saturable manner, and was released in active anticoagulant form by plasmin but not by urokinase. The complex formation was inhibited by 5 mM calcium or 250 mM NaCl as well as by polybrene or spermin. It is suggested that fibronectin binds both heparin and collagen cooperatively to form an insoluble ternary complex of the extracellular matrix.     

On the effect of the food ofMyzus persicae Sulz. on the hymenopterous parasiteAphelinus asychis Walker

Summary The effect of different synthetic diets of the hostMyzus persicae on the parasiteAphelinus asychis was tested. Deficient diets were prepared by reducing sucrose or iron concentration: sucrose concentration was reduced from 15% in the complete diet (D.I.) to 7.5% in D. II and 3.75% in D. III; that of iron from 0.0023% in D. I to 0.0011% in D. IV and 0% in D.V.The development of the immature stages ofA. asychis in aphids fed on the deficient diets was delayed (especially on D. V). The size of the emerging adults was small, which corresponds with the reduced size of the host. The percentage of the male parasites was found to be higher on the deficient diets than on the full one. The lowest average longevity of the females was found among those reared on aphids on D. III and D.V.If honeydew fromM. persicae was offered to the females, a marked reduction in the longevity of these parasites was observed on D. II and D. III. On the other hand, on honeydew from aphids on D. IV and D.V,A. asychis lived as long as on honeydew from those fed on D.I.The fecundity of the females ofA. asychis was found to be dependent on the food composition of the hosts during larval development as well as during adult life.     

Effect of past stress on the resistance of the myocardium to hypoxia

The effect of emotional painful stress on myocardial contractility and resistance to hypoxia was studied on the rat isolated atrium. It was established that in stress-exposed rats, myocardial resistance to hypoxia was reduced and contractility was depressed. It was manifested in accelerated development and greater degree of hypoxic contracture, as well as in a slower recovery of myocardial contractility under reoxygenation. The decreased myocardial resistance to hypoxia under stress is suggested to be related to the stress-induced alterations in glycolysis and calcium transport in cardiomyocytes.     

Medicine at the centre of the nation's affairs.

The medical profession was shaped a century or so ago by the interaction of three forces. These were a class structure in which doctors were largely held to be gentlemen to whom deference was due, a society dominated by the activity of production (hence the label of the working class for the majority), and the doctrines of liberalism as the guiding star in politics. Prime Minister Gladstone defined his task as "opening doors and windows." The outcome was a minimum of government interference and control with the belief that professional self regulation was the way to ensure that practice matched principle. The state (the word was hardly ever used) was self effacing almost to the point of non-existence. These are ghosts of the past, but it is a comparatively recent past. Within memory, the major domestic preoccupation of politicians of all parties was how, and to what ends, the working class could be absorbed into the political system. Health had a crucial part to play in this task, as Lloyd George and others saw early on. The "panel" was very much a forerunner of the NHS. Indeed, Bevan based part of his case in 1946 on the claim that 21 million people were already on the "panel," clear evidence of the degree to which society was still dominated by production. While speaking of Nye Bevan, we might examine his claim that the NHS was "pure socialism." In fact, it was rather closer to being "impure liberalism" in the consideration with which general practitioners and consultants were treated, the considerable freedom enjoyed by local administrations, and the low profile of government itself. That is why many remember the period as something of a golden age. It suited almost everyone very well.     

Purification of the extracellular pectinolytic enzyme from the fungus Rhizopus oryzae NBRC 4707

The pectinolytic enzyme from the solid-state culture of Rhizopus oryzae NBRC 4707 was purified to homogeneity by column chromatography on CM-Toyopearl 650 M and hydroxylapatite. The molecular weight of the enzyme was estimated by SDS-polyacrylamide gel electrophoresis to be 31,000 and was reduced to 29,700 after treatment with endoglycosidase H. Maximal activity was observed near pH 4.5 at 45°C. The enzyme was shown to be endopolygalacturonase, as judged from the formation of oligogalacturonides as its reaction products. The addition of purified enzyme, as expected, enhanced the formation of lactic acid and ethanol in potato pulp grown with R. oryzae.     

Health status of the stands and the young growth in the native spruce forests at the ural foothills,the upper reaches of the Pechora River

Health status of trees was studied in five types of native spruce forests in the Pechora-Ilych Nature Biosphere Reserve. The study has shown that trees in the old-growth bilberry-spruce and fern-shamrock-spruce forests can be characterized as “healthy”, with damage index varying from 0.03 to 0.5. The haircap-moss spruce forest with damage index 0.7 is categorized as a “weakened stand”. Regeneration was pronounced in all forest types. Young growth (1500 to 4300 stems/ha) is mainly composed of conifers and is classified as “healthy”.     

Morphology of the female reproductive organs of the African lion (Panthera leo)

The topography and splanchnology of the reproductive organs of the African lioness were studied and described in situ and after removal. The kidneys were located far caudally in relation to the thirteenth ribs. The suspensory ligament was very well developed, originated in a fan‐like manner from the dorsolateral abdominal wall lateral to the kidney and extended up to a few centimetres cranial to the kidney. The proper ligament of the ovary as well as the round ligament was well developed. The round ligament inserted on the medial femoral fascia. The left ovary was bigger than the right. The ovarian bursa had a short mesosalpinx that did not cover any part of the ovary and the fimbriae extended along the entire length. The urethral tuberculum as well as the urethral crest were well developed. The left uterine horn was longer than the right. The uterine tube was found to open directly into the tip of the uterine horn and not onto a papilla. The reproductive organs of the lioness resembled those of the domestic cat and dog but with some major differences.     

Purification of the colicin I receptor

The colicin I outer membrane receptor was solubilized from the cell envelope of Escherichia coli K12 by extraction with Triton X-100 and purified to homogeneity by a combination of ion exchange and gel filtration chromatography as well as isoelectric focusing. The receptor was isolated as a single polypeptide and retained capacity to form a complex with pure colicin. The apparent molecular weight of the receptor as determined by polyacrylamide gel electrophoresis in sodium dodecy sulfate was 74,000 or 54,000 depending on whether the preparation was boiled or not in sodium dodecyl sulfate, respectively, prior to electrophoresis. Isoelectric focusing of the receptor in the presence of Triton X-100 revealed that the protein was slightly acidic (pI 4.75).     

Factors influencing the oxidation of the radioprotector WR-1065

N-(2-Mercaptoethyl)-1,3-diaminopropane (WR-1065) is the free thiol form of the radio- and chemoprotector S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721). Interest currently exists in the clinical use of WR-2721 and WR-1065 as radio- and chemoprotectors of normal tissues. However, measurement of plasma levels of WR-1065 has proven difficult, due to rapid drug oxidation. Therefore, we studied factors influencing the oxidation of WR-1065, in Hepes-buffered saline as well as in tissue culture media containing 10% fetal bovine serum. The rate of oxygen consumption by WR-1065, as determined using the Clark oxygen electrode system, was faster in medium plus serum than in Hepes-buffered saline. That this effect is largely due to the presence of trace metal ions in tissue culture media and serum was indicated by the observation that addition of Cu2+ or Fe3+ to buffer stimulated oxygen consumption. Addition of KCN inhibited the reaction of WR-1065 with oxygen, and this effect was dependent on KCN concentration. That KCN blocked WR-1065 oxidation to the disulfide was verified using Ellman's reagent to quantitate the free thiol form. The rate of oxygen consumption was shown to be affected by temperature as well as concentration of WR-1065. Catalase reduced the rate of oxygen consumption of WR-1065, indicating that peroxide is formed in this system. Superoxide dismutase had a stimulatory effect. WR-1065 was found to stimulate the hexose monophosphate shunt in A549 cells. Since this stimulation was prevented by the presence of catalase, it appeared to be due to the response of the cells to peroxide, formed as a result of WR-1065 autooxidation.     

The development of the pouch-like structures at the angles of the mouth in the ontogeny of the hamster Phodopus campbelli]

The development of mouth angle sacciform structure was studied in the ontogenesis of hamster Phodopus campbelli. It was found that the true sacciform structure with all components characteristic for adult animals was formed by the 20th day of postnatal development. Histological data have shown than the sacciform structure is formed in ontogenesis as a result of a complex epidermal transformation involving muscular and connective tissues which comprise the external coat. The structure studied is not formed on the basis of sebaceous glands and cannot be assigned to the class of gland as considered before.