The effect of sodium salicylate on the osteoclast-like cell formation and bone resorption in a mouse bone marrow culture

Salicylates are reported to have an inhibitory effect on bone resorption in vivo and in vitro. The present study examined the effect of sodium salicylate on the formation of osteoclast-like cells in vitro. When mouse bone marrow cells were cultured for 8 days with 10^-8 M 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃), numerous clusters of mononuclear and multinucleated cells (MNCs) formed, which stained positive for tartrate-resistant acid phosphatase (TRAP-positive). In similar cultures using sodium salicylate, the number of both TRAP-positive mononuclear and TRAP-positive MNCs were found to diminish in proportion to the concentration of sodium salicylate. A time-course experimental model showed that the number of TRAP-positive MNCs decreased slightly when sodium salicylate was given early in the culture period, and decreased markedly when the drugs were given later in the culture period. Pit formation and bone-resorption area on the bone slices were also inhibited by adding sodium salicylate continuously with 1,25(OH)₂D₃. The sodium salicylate showed no cytotoxic effect because the total number of adherent cells, including both TRAP-positive and TRAP-negative cells, was independent of the presence of sodium salicylate. These results suggest that sodium salicylate has an inhibitory effect on the recruitment of osteoclast-like MNCs and that this inhibition is greater during the later stage of mouse bone marrow culture.     

An in vitro method for comparing the efficacy of two preservation solutions in one canine liver using the 5′-nucleotidase assay

The activity and localization of the plasma membrane-bound enzyme 5-nucleotidase (5-NT) in liver tissue are sensitive parameters of ischemic damage. The value of 5-NT as a marker of liver graft viability was studied in relation to liver preservation. In six mongrel dogs, the main right and left branches of the portal vein were cannulated and flushed separately in situ with cold University of Wisconsin (UW) solution and Euro-Collins (EC) solution, respectively. After hepatectomy, the right and left liver lobes were split and stored at 5°C in either of the two solutions. 5-NT activity was demonstrated in cryostat sections of liver tissue using the lead salt method. After 48 h of storage in EC solution, the 5-NT score had decreased to 31%±16% (n=6), whereas in UW solution the 5-NT score was 76%±10% (n=6). Significantly (P<0.05) higher 5-NT scores were also found after 24-h and 72-h preservation times in UW versus EC solutions. This result is in keeping with the higher preservation tolerance of liver grafts preserved in UW solution. The 5-NT assay was studied in relation to graft function in orthotopic liver transplantation experiments in dogs. All dogs with liver grafts preserved in UW solution for 24 h (n=4) and 48 h (n=3) survived (>5 days). pretransplant 5-NT scores ranged from 61% to 100%. The 72-h-preserved livers (n=5) did not show life-supporting function. Pretransplant 5-NT scores (33%±12%, n=5) were significantly (P<0.05) decreased. The 5-NT score pretransplantation was a more reliable indicator of graft function than peak SGOT values post-transplantation. In conclusion, the 5-NT assay, in conjunction with the double flush method through the portal vein, provides a simple and rapid in vitro method to test solutions for liver preservation.     

The effect of phenytoin on parathyroid extract and 25-hydroxycholecalciferol-induced bone resorption: Adenosine 3′, 5′ cyclic monophosphate production

Phenytoin, at a concentration which is known to inhibit parathyroid extract-induced bone resorptionin vitro, has been shown to have no significant effect on either the cyclic AMP content of four-day-old mouse calvaria which had been in tissue culture for 48 h or on the increase in cyclic AMP content occurring after the addition of parathyroid extract to the medium. Phenytoin has also been shown to be an effective inhibitor of bone resorptionin vitro induced by 25-hydroxycholecalciferol. These observations suggest that the inhibitory effect of phenytoin on parathyroid extract and hormone-induced bone resorption takes place beyond the cyclic AMP stage.     

Effect of 1,25-dihydroxycholecalciferol on adenosine 3′,5′-cyclic monophosphate production in calvaria of mice

Summary 1,25-dihydroxycholecalciferol (1,25-(OH)₂D₃), added at doses which stimulate bone resorption in the system used, did not increase adenosine 3,5-cyclic monophosphate (cAMP) production in vitro in mouse calvaria incubated for up to 48 h. Thus 1,25-(OH)₂D₃ does not appear to stimulate bone resorption through involvement of an increased cAMP production.     

The effect of pyrophosphate infusion on the response of the thyroparathyroidectomized rat to parathyroid hormone and adenosine-3′,5′-cyclic monophosphate

The infusion of inorganic pyrophosphate at a rate of 15 moles/h into thyroparathyroidectomized rats blocked the effects of parathyroid hormone infusion upon plasma calcium and phosphate, and upon the renal excretion of calcium, phosphate, hydroxyproline-containing peptides, and cyclic 35 AMP. In contrast, pyrophosphate infusion caused no significant change in the effects of cyclic 35 AMP infusion upon plasma calcium and phosphate, or upon the urinary excretion of calcium, phosphate, and hydroxyproline-containing peptides.

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Zusammenfassung Wurde thyroparathyreoidektomierten Ratten anorganisches Pyrophosphat mit einer Geschwindigkeit von 15 Mol/Std infundiert, so blockierte dies die Wirkung der gleichzeitig laufenden Parathormoninfusion auf Plasmacalcium und-phosphat, sowie auf die renale Ausscheidung von Calcium, Phosphat, Hydroxyprolin-haltigen Peptiden und cyclischem 3,5-AMP. Hingegen bewirkte die Pyrophosphatinfusion keine signifikante Veränderung der Wirkung einer Infusion von zyklischem 3,5-AMP auf Plasmacalcium und-phosphat oder auf die Ausscheidung im Urin von Calcium, Phosphat und Hydroxyprolin-haltigen Peptiden.

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Résumé L'injection de pyrophosphate inorganique, à la concentration de 15 moles/h, au niveau de rats thyro-parathyréodectomisés, inhibe les effects de l'hormone parathyroidienne injectée, sur le calcium et le phosphate plasmatique et sur l'excrétion rénale du calcium, du phosphate, des peptides contenant l'hydroxyproline et de l'AMP 35 cyclique. Inversement, l'injection de pyrophosphate ne provoque pas de changements significatifs sur les effects d'injection d'AMP 35 cyclique sur le calcium et le phosphate du plasma, ou sur l'excrétion urinaire de calcium, phosphate et peptides contenant de l'hydroxyproline.

     

Atrial natriuretic peptide and cyclic 3′5′-guanosine monophosphate as indicators of fluid volume overload in children with chronic renal failure

Plasma atrial natriuretic peptide (ANP) and cyclic 35-guanosine monophosphate (cGMP) were investigated as indicators of fluid volume overload in children and adolescents with chronic renal failure. Plasma ANP and cGMP were measured in both paediatric patients with chronic renal failure (n=17, mean serum creatinine 371±242 mol/l) and those with end-stage renal disease on haemodialysis (n=18). cGMP was higher in children with chronic renal failure than in 45 healthy controls (1.0±0.4 vs 2.1±0.8 nmol/l,P<0.01), whereas plasma ANP was similar (26.9±9.7 vs 34.0±12.3 pmol/l). Both ANP and cGMP were markedly elevated in children with end-stage renal disease before haemodialysis and fell significantly during dialysis. During dialysis body weight decreased by 1.6±0.7 kg, corresponding to 4.5±2.1% of body weight. Plasma ANP correlated positively with plasma cGMP in haemodialysed patients (r=0.43,P<0.05). Reduction in body weight and in mean arterial pressure correlated more closely with plasma ANP than with cGMP. Therefore, elevation of plasma ANP appears to indicate volume overload in children undergoing haemodialysis, but whether it can be used also in children with chronic renal failure requires further investigation     

Testosterone metabolism in primary cultures of epithelial cells and stroma from benign prostatic hyperplasia

We studied the metabolism of testosterone in primary cultures of prostate epithelial cells and fibroblasts obtained from patients with benign prostatic hyperplasia (BPH). The conversion of ³H-testosterone in both cell cultures was predominantly to the oxidative pathway, with the formation of ³H-androstenedione increasing with cell number and time of incubation. Although we also detected some 5-reductase activity in these cells, the activity in the stroma component (0.00688 pmol/mg protein/min) was nonetheless insignificant when compared to the 5-reductase activity in the tissue of origin (0.0616 pmol/mg protein/min) and well below the 17-hydroxysteroid dehydrogenase activity of the same cells (0.0518 pmol/mg protein/min). The aromatase activity in our cells was also measured by two separate techniques, but neither the deuterium procedure nor the production of oestrone from androgen precursors yielded any positive results, suggesting that under these experimental conditions there was no aromatase activity within the cells. The shift from the reductive to the oxidative pathways in these primary cell cultures was reminiscent of the androgen-metabolizing enzyme profiles seen in poorly differentiated prostate cancer. Whether this fransition is an obligatory step in the development of hormone refractiveness remains to be elucidated.     

A mechanism for topical cutaneous calcinosis

ATP has been found in the fluid from distilled water washes of the dorsum of epilated and non-epilated rats. However, considerably more ATP was found in the epilated rats. Based upon these and previously reported results demonstrating that calcium adenosine 5 triphosphate initiates subcutaneous calcification in rats, a hypothesis is proposed for the mechanism of topical cutaneous calcinosis as follows: Feeding DHT results in generalized hypercalcemia. When this is followed by epilation, tissue trauma occurs, resulting in destruction and/or enhanced permeability of cell membranes and release of ATP. The CaATP which forms hydrolyzes to produce a calcium orthophosphate nucleating seed for the calcification process. The ground substance necessary to form with calcium phosphate, the organized plaque, are the collagen fibers and muccopolysaccarides in the inter-fibrillar matrix in the dermal layer or the hair follicle itself.

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Zusammenfassung Das Dorsum von enthaarten und nichtenthaarten Ratten wurde mit destilliertem Wasser gespült. In dieser Flüssigkeit wurde ATP festgestellt. Bei enthaarten Ratten wurde jedoch beträchtlich mehr ATP gefunden. Auf Grund dieser sowie früher mitgeteilter Befunde, wonach Calcium-Adenosin-5-Triphosphat die subkutane Verkalkung bei Ratten auslöst, wird folgende Hypothese für den Mechanismus der lokalen Hautcalcinosis vorgeschlagen. DHT-Verabreichung führt zu generalisierter Hypercalcaemie. Darauffolgende Enthaarung führt zu Verletzung des Gewebes, was die Zerstörung und/oder erhöhte Permeabilität der Zellmembranen und Abgabe von ATP nach sich zieht. Das dadurch entstehende, CaATP wird hydrolysiert und dabei entsteht ein Calciumorthophosphat-Nukleationskern für den Verkalkungsvorgang. Die Grundsubstanz, welche nötig ist, um mit Calciumphosphat diese Plaque zu bilden, besteht aus Collagenfasern und Mucopolysacchariden der inter-fibrillären Matrix der Hautschicht oder aus dem Haarfollikel selbst.

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Résumé Del' ATP a été trouvé dans les lavages à l'eau distillée du dos épilé et non-épilé de rats. Cependant, des quantités plus élévées d'ATP sont observées chez les rats épilés. En fonction de ces résultats et de travaux antérieurs démonstrant que l'adénosine 5 triphosphate de calcium provoque une calcification sous-cutanée chez le rat, une hypothèse concernant le mécanisme de la calcinose cutanée, locale est proposée. L'ingestion de DHT provoque une hypercalcémie généralisée — Si une épilation est ensuite réalisée, un trauma tissulaire se developpe, avec destruction et/ou perméabilite augmentée des membranes cellulaires et rejet d'ATP. Le Ca ATP qui se forme, s'hydrolyse pour constituer une zone de nucléation pour le processus de calcification. La substance, fondamentale, nécessaire pour la formation de la plaque organisée, est constitutuée par les fibres de collagène et les mucopolysaccharides de la matrice interfibrillaire de couche dermique du follicule pileaux lui-même.

     

Fibrinogen osaka IV: A congenital dysfibrinogenemia found in a patient originally reported in relation to surgery,now defined to have an Aα arginine-16 to histidine substitution

We discovered a congenital heterozygous dysfibrinogen in a patient and reported this case in relation to surgery some time ago (Jpn J Surg (1988) 18:43–46).³ Further studies on the isolated abnormal population of fibrinogen derived from this patient have revealed that fibrinopeptide A was not cleaved by ancrod, a snake venom-derived thrombin-like enzyme, but by thrombin, slowly but completely. The released fibrinopeptide A components, being the A, AY, and AP peptides, were all found to be abnormal, as evidenced by slightly earlier elution positions on high-performance liquid chromatography, compared with the normal counterparts. By analyzing their amino acid sequence, we have identified an arginine to histidine substitution at position 16 of the A chain, the thrombin cleavage site. Utilizing insolubilized abnormal fibrinogen, we confirmed that the polymerization site assigned to the central E domain, the A site, was exposed by thrombin, but not by ancrod. This dysfibrinogen, designated as fibrinogen Osaka IV, is the second abnormal molecule with an A arginine-16 to histidine substitution identified among Japanese families.     

Das Krebsrisiko im Resektionsmagen

Zusammenfassung Es wurde die Frage geprüft, ob obligater duodenogastrischer Reflux unter Langzeitbedingungen das Wachstum von Carcinomen oder Sarkomen im resezierten Magen beeinflußt. 72 männliche Wistarratten wurden einer Magenresektion unterworfen. Die erforderliche Gastroenteroanastomose wurde bei 39 Tieren als short loop GE (Billroth II) mit obligatem Reflux von Galle und Duodenalsekret in den Restmagen angelegt. Die restlichen 33 Tiere erhielten eine Y-förmige GE (nach Roux) mit seitlicher Einmündung des Duodenum in den Dünndarm; Reflux von Galle in den resezierten Magen war dadurch unmöglich. Die Carcinominduktion erfolgte durch kontinuierliche Gabe von N-Methyl-N'-Nitro-N-Nitrosoguanidin im Trinkwasser. Bei der Sektion der Tiere mit Reflux 33 Wochen nach Carcinogengabe fand sich die Mehrzahl der malignen Tumore von Magenstumpf, Anastomose (GE) und angrenzendem Dünndarm in der Tiergruppe mit obligatem Reflux (13 von 22). Die Tiergruppe ohne Reflux (Y-Roux GE) wies demgegenüber signifikant weniger Carcinome auf (7 von 21); die Tumorincidenz dieser Gruppe entsprach derjenigen eines nicht operierten Kontrollkollektives (5 von 22). - Die Ergebnisse unserer Untersuchungen zeigen, daß die in der Technik des short loop durchgeführte Gastroenteroanastomose des Billroth 11-Magens das Krebswachstum im Anastomosengebiet in besonders ausgeprägtem Maße fördert. Ausschlaggebend für die gesteigerte Carcinominduktion ist der chronische Reflux von Galle und Duodenalsekret, der bei der short loop-GE obligat über den Magnrest fließt.

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Cancer risk of the stomach resected for ulcerThe role of duodenogastric reflux

Summary The purpose of this study was to find whether the reflux of bile and pancreatic juices following stomach resection (duodenogastric reflux) enhances the incidence of carcinomas near the gastroenteric anastomosis. 72 male Wistar rats were subjected to stomach resection. The gastroenteric anastomosis (GE) was performed either as short loop anastomosis (Billroth II;n = 39), thus providing a continuous duodenogastric reflux, or as Y-shaped GE (according to Roux;n = 33). By the latter technique, bile and pancreatic juices are derived quantitatively into the jejunum without coming into contact with the remaining part of the stomach. During a period of 33 weeks, operated rats as well as intact animals were given the carcinogen N-methyl-N-nitro-N-nitrosoguanidine (NG; 120 mg/1) in the drinking water. At autopsy, 33 to 36 weeks after daily oral administration of NG, most of the tumours were found in operated rats undergoing continuous ref lux (Billroth II group). In contrast to these findings, the incidence of carcinomas was significantly lower in animals without reflux (Roux group or intact control rats).-The results of our experiments demonstrate that, in rats, the duodenogastric reflux contributes substantially to the development of carcinomas of the resected stomach.

Mit Unterstützung der Deutchen Forschungsgemeinschaft     

The role of the coagulation cascade in brain edema formation after intracerebral hemorrhage

Summary The coagulation cascade has a potential role in brain edema formation due to intracerebral hemorrhage. In this study blood and other solutions were injected stereotactically into the right basal ganglia in rats. Twenty-four hours following injection, brain water and ion contents were measured to determine the amount of brain edema. Intracerebral blood resulted in an increase in brain water content. The amount of brain edema surrounding the intracerebral hematoma was reduced by a thrombin inhibitor Na-(2-Naphthalenesulfonylglycyl)-4-amidino-DL-phenylalaninepiperidide, (-NAPAP) infused into the hematoma after the clot had been allowed to solidify. The inhibitor did not alter the actual size of the clot mass. An artificial clot composed of fibrinogen, thrombin, and styrene microspheres also produced brain edema. A fibrin clot led to edema formation even in the absence of mass effect provided by the microspheres. The single component responsible for production of brain edema in all these models was thrombin. The edema was formed in response to a fibrinogen-independent pathway. These results indicate that the coagulation cascade is involved in brain edema that develops adjacent to an intracerebral hematoma.     

Analgesic effect of intravenous ATP on postherpetic neuralgia in comparison with responses to intravenous ketamine and lidocaine

Purpose No study has been performed on the analgesic effect of adenosine 5-triphosphate (ATP) on postherpetic neuralgia (PHN). We conducted an open-label trial of ATP in patients with PHN, and compared ATP with ketamine and lidocaine.Methods Twelve patients with PHN were studied. On separate days, ketamine (0.3mg·kg^–1), lidocaine (2mg·kg^–1), and ATP (100µg·kg^–1·min^–1 or less for 120min) were administrated intravenously. The intensity of spontaneous pain as well as tactile allodynia was assessed using a visual analog scale (VAS). When the VAS score for spontaneous pain was decreased by more than 50%, the patient was classified as a responder.Results Five, 6, and 6 patients responded to ketamine, lidocaine, and ATP, respectively. In 6 ATP responders, pain relief developed slowly and lasted for 9 (median) h (range: 3–72h). All 5 ketamine responders and only 1 of 7 ketamine nonresponders responded to ATP (5/5 vs 1/7, P < 0.05, ² test) whereas 2 of 6 responders to lidocaine and 4 of 6 nonresponders to lidocaine responded to ATP (2/6 vs 4/6, P > 0.05). The ketamine responders responded to ATP more often than did the lidocaine responders (5/5 vs 2/6, P < 0.05).Conclusion Intravenous ATP exerted slowly developing and long-lasting analgesic effects in half of patients with PHN. Patients with ketamine-responsive PHN were likely to respond to ATP.     

Vitamin C und Dünndarmkrebs

Zusammenfassung Die Studie gilt der Wirkung von Vitamin C auf die chemisch induzierte Dünndarmcarcinogenese der Ratte. Durch Verabreichung von N-Äthyl-N-Nitro-N-Nitrosoguanidin (ÄNNG) im Trinkwasser (120 mg/l) wurden nach 18 Wochen bei über 90 % der Tiere Dünndarmtumoren erzeugt. Diese Tumorinduktion konnte durch die orale Verabreichung großer Mengen von Natriumascorbat im Futter (2g/100g Futter) nicht unterdrückt werden. Allerdings wird die Tiefeninfiltration der erzeugten Carcinome gehemmt. So haben 29 von 36 Ratten, die ausschließlich ÄNNG erhielten, ein Carcinom entwickelt. 25 Tiere (69%) dieser Gruppe wiesen ein Tumorstadium P₄ auf. Hingegen wurde bei 24 von 35 Tieren, die zusätzlich Vitamin C erhielten ein Carcinom beobachtet, das aber nur bei 13 Tieren (37 %) das Tumorstadium P₄ erreichte. Die alleinige Verabreichung von Vitamin C erzeugt keine Dünndarmveränderungen.

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Ascorbic acid and cancer of the duodenumAn experimental study

Summary This study applies to the agency of vitamin C on chemical carcinogenesis in the small intestine of rats. Administration of N-Ethyl-N-nitro-N-nitrosoguanidine (ENNG) in drinking water produced tumors of the small intestine after 18 weeks in more than 90 %. The induction of tumors could not be suppressed by a large amount (2–3 %) of sodium ascorbate in food, but the depth of tumorinfiltation was restricted. Cancer developed in 29 of 36 rats receiving ENNG only. In 25 animals of this group growth of tumor corresponded to a P₄ stage. In 24 of 35 animals, additionally receiving vitamin C, P₄ stage was observed in only 13 cases. Pathological changes in the small intestine could not be observed after the sole administration of vitamin C.

     

Some remarks on breast reduction by the “B” technique

The main disadvantage of the popular techniques for breast reduction are the scars left around the breasts. We present our experience with the B technique for breast reduction, which creates a much smaller size scar that is well hidden under the brassiere. Our conclusion is that the B technique is the method of choice for most breast reductions.     

Changes in the productivity of cytokines and active-oxygen in peripheral blood cells following surgery

An investigation was carried out on the productivity of cytokines and active-oxygen by peripheral blood cells during the pre- and post-operative periods. While the preoperative production of tumor necrosis factor (TNF) and interleukin-l (IL-1) was elevated, that of lymphotoxin (LT) and interferon- (IFN-) were slightly suppressed. In the postoperative period the peak TNF and IL-1 and active-oxygen productivity was elevated, while LT and IFN- productivity was suppressed in patients with an intraoperative bleeding volume of more than 1,000 ml compared to those with that of less than 1,000 ml. Thus, stress stimulates the TNF and IL-1 and active-oxygen producing system, that is, the macrophage-neutrophil system, and suppresses the LT and IFN- system, being, the inflammatory helper T cell system, in the early postoperative period.     

Association of meningiomas with dural “tails”; Surgical significance

Summary Intracranial meningiomas are characteristically benign tumours with a tendency to recur following surgical resection. Our group is investigating the pathogenesis of meningioma recurrence. In our initial studies we identified two cases of dural tails associated with intracranial meningiomas. Gadolinium-enhanced magnetic resonance images were utilized to identify the dural tails preoperatively. These images aided us in performing a more complete surgical resection of the meningiomas. Histopathological confirmation of meningotheliomatous cell infiltration into the dural tails demonstrates their surgical significance.     

The diagnosis and treatment of the ‘mixed’ type of depressed fractures of the calcaneum

Summary The mixed type of depressed fractures of the calcaneum is described and a classification proposed. During a six year period, 78 patients with fractures of the calcaneum were seen. Of these 32 were of the mixed type and 23 were treated by sub-talar arthrodesis and restoration of the calcaneal outline. Very good or good results were obtained in all patients subjected to operation.

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Résumé L'auteur décrit un type de fracture du calcanéum par enfoncement «mixte» (i. e. à la fois vertical et horizontal) et il en propose une classification. Sur 78 fractures du calcanéum observées en six ans, 32 étaient de ce type «mixte» et 23 d'entre elles furent traitées par arthrodèse sous-astragalienne avec reconstitution de la forme du calcanéum. D'excellents ou de bons résultats ont été obtenus chez tous les blessés qui ont bénéficié de cette intervention.