Ultrastructural alterations in gizzard smooth muscle of selenium-vitamin-E-deficient ducklings.

Lesions of gizzard smooth muscle were studied by light and electron microscopy in newly hatched ducklings fed a selenium-vitamin-E-deficient diet for 13-21 days. Histopathologic alterations included initial hyaline change in damaged smooth-muscle cells, subsequent mineralization of sarcoplasmic debris in necrotic smooth-muscle cells, macrophagic invasion and phagocytosis of sarcoplasmic debris, and eventual fibroblastic proliferation and scarring of damaged areas in the gizzard wall. Ultrastructurally, mild damage in smooth-muscle cells was manifested by altered mitochondria with matrical densities and disrupted membranes and dilated elements of sarcoplasmic reticulum. In smooth-muscle cells with severe injury, the alterations of mitochondria and sarcoplasmic reticulum were accompanied by myofibrillar lysis and disruption of plasma membranes and external laminae. Expanding zones of mineralization of sarcoplasmic debris in necrotic smooth muscle cells were present at dense masses of lysed myofilaments that surrounded mineralized and disrupted mitochondria. Numerous macrophages infiltrated the areas of necrosis of smooth muscle and phagocytosed sarcoplasmic debris.


Fowl cholera: immunization of chickens with potassium thiocyanate (KSCN) extract of Pasteurella multocida serotype 3.

Potassium thiocyanate (KSCN) extract of Pasteurella multocida, serotype 3, was found to be immunogenic against a challenge infection of homologous as well as one heterologous strain tested (serotype 1). Ouchterlony's gel-diffusion analysis revealed the presence of two components in the KSCN-extracts of both serotypes 3 and 1, which were antigenically identical.


Photosensitizing effects of Cymopterus watsonii and Cymopterus longipes in chickens and turkey poults.

Two-week-old straight-run white broiler chicks and 2-week-old straight-run white turkey poults were fed seed or whole plants Cymopterus watsonii or seed of Cymopterus longipes at 0.4, 0.8, and 1.6% of body weight, and then placed in sunlight for 5 hours each day for 7 days. All plant material produced moderate to severe photosensitivity in chicks. Toxic signs included photophobia; red discoloration of beak, comb, and feet; loss of feathers in the periorbital area; dried serous fluid on comb and edge of beak; keratoconjunctivitis; and multiple lesions on feet and legs. Turkey poults showed a syndrome similar to that of chicks. Mortality was particularly high in turkeys fed C. longipes seed. Toxic signs in turkeys 6 weeks after treatment included tremors in the legs; upturning and shortening of the upper beak; and shortening disfigurement, and gangrene of the toes. Two phototoxic furocoumarins, oxypeucedanin and isoimperitorin, were isolated from C. longipes seed. Either compound, administered orally, produced photosensitivity in 2-week-old chicks.


Antibody response to strain combinations of Newcastle disease virus as measured by hemagglutination-inhibition.

Differences in antibody response to three Newcastle disease virus (NDV) strains--B-1, LaSota, and Ulster--were investigated using the hemagglutination-inhibition (HI) micro-titer test in chickens hatched from ND-immune and unimmune flocks. When used singly as primary vaccines, the Ulster strain stimulated the lowest antibody response of the three in both immune and unimmune (susceptible) chickens. Subgroups of each of the primary-vaccinated groups were revaccinated with each of the three strains. Ulster-vaccinated chicks, revaccinated with Ulster, gave the poorest booster response. All other revaccination combinations gave a significant titer increase, though some were better than others. It is suggested that the Ulster strain as primary vaccine followed by booster does of B-1 or LaSota will induce a higher antibody response (i.e., immunity) in susceptible chicken populations with less risk of a post-vaccination reaction.


Preincubation dipping of turkey hatching eggs. I. Effect of shell treatment on amount and variability of fluid intake.

Two techniques for egg-shell treatment were evaluated for their effects on the quantity and variability of the fluid absorbed by turkey hatching eggs subjected to the temperature-differential method of egg dipping. The use of pin or drill to pierce a uniform hole through the shell and membrane(s) on the large end of each egg caused a significant increase in the quantity of dip solution absorbed. Variability in fluid uptake by the eggs also was reduced significantly. Utilization of dilute acid to remove cuticle completely from the whole egg surface or partially from either end also had similar effects. The techniques described also permit and effective dipping procedure that does not require the preheating step.


Use of the Nobuto blood-sampling paper strip for Newcastle disease serology.

Dried whole blood collected by the paper-strip method of Dr. Kenzo Nobuto was evaluated for use in the Newcastle disease hemagglutination-inhibition (HI) test. When dried blood on the Nobuto strip was eluted in 1.0 ml of diluent, the resulting HI titers were similar to titers obtained with sera diluted 1:10. Both storage time and temperature influenced the stability of HI activity in dried blood, though the influence was minimal and is considered to have little significance in diagnostic serology, The advantages of this method of blood collection for Newcastle disease HI tests are discussed.


Interactions between viscerotropic velogenic Newcastle diseases virus and pet birds of six species. I. Clinical and serologic responses, and viral excretion.

Clinical and serologic responses to a psittacine isolate of viscerotropic velogenic Newcastle disease virus (VVNDV) were evaluated in pet birds of six species: budgerigar, yellow-headed Amazon parrot, halfmoon conure, lesser hill mynah, black-headed nun, canary. The clinical response was most marked in the budgerigars, parrots, and conures, and only minimal in the nuns. Between post-exposure days (PED) 3 and 5 some birds developed ruffled plumage, conjunctivitis, and central nervous system dysfunction: ataxia, wing tremors, paralysis of the extremities, and tremors of the head accompanied by nodding and jerking. Mortality by PED 203 was 55% (29/52) in the halfmoon conures, 22% (23/105) in budgerigars, 29% (12/42) in parrots, and 21% (15/71) in nuns. The only clinical signs in canaries and mynahs were progressive death losses, respectively 25% (33/132) and 21% (10/48). The visceral lesions common in chickens with VVNDV were not observed in these six species. Canaries rapidly eliminated Newcastle disease virus (NDV), whereas it was detected for protracted periods in the oral and cloacal secretions of the other five species (for more than a year in parrots). Serologic evaluation by the hemagglutination-inhibition and neutralization tests also indicated prolonged NDV infections in 5 of the 6 species. The seroconversion rate observed in canaries was minimal (13%).


Interactions between viscerotropic velogenic Newcastle disease virus and pet birds of six species. II. Viral evolution through bird passage.

Following in vivo studies in pet birds of 6 species, 279 Newcastle disease virus (NDV) reisolates were selected for characterization by the embryonated-chicken-egg mean-death-time, plaque-assay, hemagglutination-elution, and hemagglutinin-thermostability techniques. Initially, the 279 isolates were screened by the mean-death-time and plaque-assay techniques, and 5 sequential isolates were chosen for each of 3 budgerigars and 2 parrots for characterization by the other 2 in vitro assays to determine whether the Colorado Psittacine Isolate of viscerotropic velogenic (VV) NDV (COPI-VVNDV) had evolved during passage through pet birds. Nineteen isolates were then selected for chicken back-passage studies. Fifteen of the 19 isolates were chosen for potential avirulence for 8-week-old domestic chickens. The 4 remaining isolates produced large red plaques when assayed and were therefore used as virulent virus controls likely to be VVNDV. Subsequent in vitro characterization of selected back-passage chicken NDV isolates demonstrated little change in the 4 parameters originally evaluated for the pet-bird isolates used for the back-passage studies. Although the psittacine isolate slowly evolved to relatively avirulent strains of NDV by passage in pet birds, reversion did not occur during the chicken back-passage studies.


Turkey sinusitis: synergism between Mycoplasma synoviae and Mycoplasma meleagridis.

Sinusitis was produced experimentally in turkeys after intrasinus inoculation with broth cultures of both Mycoplasma synoviae and Mycoplasma meleagridis. No evidence of sinusitis was observed in other turkeys exposed to each of these organisms separately- indicating pathogenic synergism with these mycoplasmas.


Purification and identification of turkey immunoglobulin-A.

Turkey immunoglobulin-A (IgA) was isolated from bile, intestinal secretions, and serum by affinity chromatography using monospecific anti-turkey IgA coupled to CNBr-activated Sepharose. The isolated immunoglobulin was antigenically distinct from IgM and IgG. The purity of IgA was demonstrated by immunodiffusion, immunoelectrophoresis, and analytical ultracentrifugation. The predominant forms of polymeric IgA in bile and intestinal secretions had respective So20w values of 16.1 and 15.2. Larger polymers (25-26S) were also present. Two molecular forms (8.5S and 17S) were found in serum. The 8.5S peak was higher than the 17S, indicating a greater concentration of 8.5 S.


[Preleukemia].

The six cases of Preleukemia (or Preleukemic States) are described. The peripheral blood was characterised with Pancythopoenia in 5 and bicytopoenia in 1 patient. Morphological alterations of Erythrocytes were present in all cases. The Bone Marrow was hypercellular in 3 and hypocellular in 3 patients. Erythropoiesis was megaloblastic or partly megaloblastic in 5 cases, with "ring" sideroblasts present in all cases. The percentage of Leukemic blast cells was as follow: 0, 5, 6, 10, 30 and 33. The duration of the preleukemic phase varied from 3-53 months. With the manifestations of overt Acute Nonlymphatic Leukemia the agressive therapy was applied. The results were poor in 5 patients (the survival 1, 5-4 m.). Only one survived more than 60 months and is still in full remission.


An outbreak of erysipelas in Coturnix quails.

A severe outbreak of erysipelas causing high mortality was observed in Coturnix breeder quails. Possible source(s) of erysipelothrix infection in the flock and subsequent infections due to Pasteurella multocida, Salmonella typhimurium, and Staphylococcus aureus are discussed.


[Substitution therapy of coagulopathies].

In this work are presented current experiences and attitudes in the substitutional therapy of acquired and congenital coagulopathies. There is accentuated general accepted attitude at present that for an adequate supstitution is necessary decided knowledge of the clinical and laboratory diagnostic of these diseases, as much as the possibility of arrangements of the specific chemo-preparations and their doses, and the frequency of application and all complications that might result in the frame of this view of therapy.


Pasteurella anatipestifer infection in turkeys.

In a case of excessive mortality in seven-week-old turkeys, the primary lesion at necropsy was severe fibrinous pericarditis and epicarditis. After 48 hours of incubation at 37 C, Pasteurella anatipestifer was isolated. The infection responded to antibiotic therapy with a combination of neomycin and oxytetracycline in the drinking water. The lesions suggested chlamydial infection, but cultural procedures for chlamydia were negative.


A fowl typhoid outbreak in a chicken breeder flock.

Fowl typhoid was diagnosed in a 1-year-old light-breed brown-egg-laying chicken breeder flock and in a 16-week-old progeny pullet flock on the same farm. Salmonella gallinarum was cultured from chickens in both flocks. Signs and lesions were typical. The disease was confined to the one farm. The source was suspected to have been a grossly contaminated well.


[Immunoelectrophoretic method for the determination of factor VIII-related antigens].

Immunological examination of F VIII related antigen gave some new information on the nature of congenital defects of F VIII, in haemophilia A and von Willenrand's disease. Besides classic method in diagnostics of von Willebrand's disease, the determination of F VIII related antigen can be used as a diagnostic criterium in distinguishing von Willebrand's disease from some mild forms of haemophilis, as well as in detection of haemophilia carrier. In addition, the study on the relationship of immunological value of F VIII related antigen and biological activity of F VIII offers more data on the possibility of detection of so-called "hypercoagulability" and states proceeding thrombosis. The method for determination of F VIII related antigen (Rocket electrophoresis--Laurel) as well as the values of F VIII in health persons of our population is described in this paper.


[ABO blood-group system in the fossil remains at the mesolithic cemetary in Vlasac (6,300-5,400 B.C.)].

By the several generally accepted methods, the authors tested the ABO blood group system of human fossil remains of the mezolit cementery of Vlasac (lower flow of the River Danube). The results of these examinations showed a biological similarity of the population in all of the three cementery layers and a difference in blood groups in these people and the present inhabitants of that region. These examinations are of great interest for the study of the structure of the prehistorical population and its migrations.


Metoserpate Hydrochloride for the treatment of hysteria in replacement pullets.

Fifteen flocks of replacement pullets affected with hysteria were treated with metoserpate hydrochloride via the drinking water after water was withheld for an adequate period. The dose was 4 mg/kg body weight (BW) on day 1, and 2 mg/kg BW on days 5 and 9. Metoserpate hydrochloride proved effective and safe for treating clinical cases of hysteria in replacement pullets.


Distribution of the Lewis blood-group system in 331 members of JNA.

The aim of this paper is to describe the phenotype distribution in Yugoslavians, members of JNA. The results are systematised into two groups: a. phenotype distribution of the blood-group system in tested persons; b. the relationship of phenotypes and ABO blood-group system.


The biosynthesis of brain gangliosides. Separation of membranes with different ratios of ganglioside sialylating activity to gangliosides.

Brain subcellular fractions were analysed for ganglioside-sialylating activity by measuring the incorporation of N-[3H]acetylneuraminic acid from CMP-N-[3H]acetylneuraminic acid into endogenous ganglioside acceptors (endogenous incorporation) and into exogenous lactosyceramide (haematoside synthetase activity). The ratios of endogenous incorporation to gangliosides and of haematoside synthetase to gangliosides for the synaptosomal and mitochondrial fractions from a washed crude mitochondrial fraction were lower than those obtained for other membrane fractions. The differences appear to reflect intrinsic characteristics of each membrane fraction. The results of labelling in vitro and the time course of labelling of gangliosides of the different subcellular fractions in vivo after injection of N-[3H]acetylmannosamine are consistent with the possibility of a subcellular site for synthesis of gangliosides different from that of ganglioside deposition.


Modifications of major aspects of myocardial ribonucleic acid metabolism as a response to noradrenaline. Action of the hormone on cytoplasmic processing of ribonucleic acid after reserpine treatment.

Treatment of perfused rabbit heart with reserpine causes a decrease of incorporation of labelled precursors into RNA species of subcellular fractions and polyamines. Ornithine decarboxylase, S-adenosylmethionine decarboxylase and cytoplasmic Mn2+-stimulated polyadenylate polymerase activities are not modified. Addition of noradrenaline to reserpine-treated perfused hearts enhances, compared with the control, the incorporation of precursor into RNA in all subcellular fractions other than the nuclear one, restores incorporation of labelled putrescine into polyamines, enhances ornithine decarboxylase and S-adenosylmethionine decarboxylase activities and causes a 12-fold increase in cytoplasmic Mn2+-dependent polyadenylate polymerase activity. After treatment with noradrenaline the increase in radioactivity was found solely in AMP after hydrolysis of microsomal RNA to nucleoside monophosphates.


Effects of hormones on the synthesis of alpha 1 (acute-phase) glycoprotein in isolated rat hepatocytes.

Hormone effects on the synthesis of alpha(1) (acute-phase) glycoprotein and of albumin by isolated rat hepatocytes in suspension were examined. Insulin, glucagon, cortisol, somatotropin (bovine growth hormone) and tri-iodothyronine were added to achieve physiological concentrations in the medium [Jeejeebhoy, Ho, Greenberg, Phillips, Bruce-Robertson & Sodtke (1975) Biochem. J.146, 141-155]. After periodic additions, there were increases (compared with values for non-hormone-treated suspensions) in the concurrent absolute syntheses of alpha(1) (acute-phase) glycoprotein and of albumin. Trends were detectable after 24h, and significant increases were demonstrated after 48h of incubation (219 and 119% respectively of control values). Manipulation of hormones, by omission from the mixture or by addition of only one or two hormones in various combinations, indicated that for alpha(1) (acute-phase) glycoprotein (which may be representative of some other acute-phase proteins), cortisol was one of the most important hormones involved in the stimulation of synthesis, with glucagon enhancing the effect of cortisol but not being stimulatory by itself. Addition of actinomycin D inhibited this stimulation, suggesting that cortisol might have acted through promotion of RNA synthesis. For albumin, cortisol alone did not stimulate synthesis, but its absence from a hormone mixture significantly decreased synthesis compared with that observed with the complete hormone mixture. Our findings support the possibility that following tissue injury, synthesis of alpha(1) (acute-phase) glycoprotein may be stimulated by the hormonal response to this injury (which response includes elevated blood concentrations of cortisol and glucagon).


Biliary excretion of some anionic derivatives of diethylstilboestrol and phenolphthalein in the guinea pig.

The metabolic fates and modes of excretion of diethylstilboestrol mono[35S]sulphate and diethylstilboestrol di[35S]sulphate were studied in the guinea pig. Comparative studies were also made with [G-3H]diethylstilboestrol and phenolphthalein di[35S]sulphate. Diethylstiboesterol di[35S]sulphate was extensively eliminated in the bile unchanged. After administration of diethylstilboestrol mono[35S]sulphate, extensive biliary elimination of radioactivity was also recorded. Radioactive components were identified as diethylstilboestrol disulphate, diethylstilboestrol monosulphate monoglucuronide and unchanged diethylstilboestrol monosulphate. When [G-3H]diethylstilboestrol was administered, 3H-labelled diethylstilboestrol monoglucuronide, diethylstilboestrol monosulphate monoglucuronide and diethylstilboestrol disulphate appeared in the bile. Phenolphthalein di[35S]sulphate was excreted unchanged in bile. These findings are discussed in relation to studies carried out in the rat [Barford, Olavesen, Curtis & Powell (1977) Biochem. J. 164, 423--430] and species differences are related to differences in enzyme activities in rat and guinea-pig liver.


Regulation of L-ornithine decarboxylase and S-adenosyl-L-methionine decarboxylase in rat ventral prostate and seminal vesicle.

1. The activities of l-ornithine decarboxylase (EC 4.1.1.17) and S-adenosyl-l-methionine decarboxylase (EC 4.1.1.50) were dramatically enhanced in both the ventral prostate and the seminal vesicle of castrated rats in response to androgenic stimulation. The time course of the stimulation of ornithine decarboxylase together with the quantitatively different response of adenosylmethionine decarboxylase to testosterone treatment in the prostate gland and seminal vesicle indicated that the enhancement in polyamine synthesis in the ventral prostate may reflect both cellular proliferation and the restoration of the secretory functions of the organ. In the seminal vesicle, however, the stimulation of the polyamine-biosynthetic pathway more closely resembled the pattern found in other rat tissues, such as regenerating liver, undergoing compensatory growth. 2. Ornithine decarboxylase activity in the ventral prostate and especially in the seminal vesicle of sexually mature rat was diminished in vivo by various short-chain diamines such as 1,2-diaminoethane, 1,3-diaminopropane and putrescine (1,4-diaminobutane). These diamines had no direct effect on the enzyme activity in vitro. 3. In contrast with the marginal decrease in ornithine decarboxylase activity produced by diaminoethane in the ventral prostate of non-castrated animals, repeated injections of the latter amine completely prevented the intense stimulation of the enzyme activity in the ventral prostate and seminal vesicle of castrated rats at 24h after the commencement of testosterone treatment. 4. The decrease in ornithine decarboxylase activity observed after injections of diamines (putrescine) in the ventral prostate was apparently associated with a similar decrease in the amount of immunoreactive protein as revealed by immunotitration of the enzyme with antiserum to rat ornithine decarboxylase.


Role of a specific choline pool in sphingomyelin synthesis in rat heart.

Sphingomyelin synthesis was studied in slices of rat heart by using [Me-14C]choline, [1,2-14C]ethanolamine, S-adenosyl-L-[14C]methionine and [32P]Pi as as precursors. In the presence of both [Me-14C]choline and [32P]Pi the ratio of the specific radioactivities of 14C and 32P in phosphatidylcholine was greater than in sphingomyelin at all the times studied. This suggested that synthesis of phosphatidylcholine and sphingomyelin de novo did not involve the utilization of a common pool of cytidine diphosphate choline. In addition, studies with [1,2-14C]ethanolamine and S-adenosyl-L-[14C]methionine indicated that a quantitatively significant pool of choline, derived from these precursors, was selectively utilized for sphingomyelin formation. This pool was not represented by phosphatidylcholine formed by methylation of phosphatidylethanolamine or by other pathways.


Porphyrins and porphyrinogen carboxy-lase in hexachlorobenzene-induced porphyria.

1. Qualitative and quantitative studies of the porphyrins and the porphyrinogen carboxylyase of the liver, spleen, kidney, harderian gland and erythrocytes from normal rats and from those hexachlorobenzene-induced porphyria were carried out. 2. Hexachlorobenzene has no effect on erythrocyte porphyrin content, but produces a decrease in that of Harderian gland and an increase in the porphyrin content of the kidney and spleen, and a marked increase in the liver (1 mumol/g of tissue). Octacarboxylic (isomer III) and heptacarboxylic porphyrins accumulated in kidney, spleen and liver, the former porphyrin being predominant. 3. Hexachlorobenzene has no effect on the activity of porphyrinogen carboxy-lase in erythrocytes; there is a slight decrease in enzyme activity in the Harderian gland, and a marked decrease in the liver and kidney enzyme activities. In the liver the removal of each carboxyl group from uroporphyrinogen III appears to be affected by this treatment. 4. The liver is the principal site of action of hexachlorobenzene, with the kidney next in decreasing order of effect, and erythropoietic tissue is unaffected. The marked decrease in porphyrinogen carboxy-lyase activities observed in liver and kidney could explain the high accumulation of octacarboxylic and heptacarboxylic porphyrins found in these tissues. 5. The results are discussed in relation to changes promoted by hexachlorobenzene in other enzymes of the haem pathway.


Effects of analogles of ethanolamine and choline on phospholipid metabolism in rat hepatocytes.

1. Analogues of ethanolamine and choline were incubated with different labelled precursors of phospholipids and isolated hepatocytes and the effects on phospholipid synthesis were studied. 2. 2-Aminopropan-1-ol and 2-aminobutan-1-ol were the most efficient inhibitors of [(14)C]ethanolamine incorporation into phospholipids, whereas the incorporation of [(3)H]choline was inhibited most extensively by NN-diethylethanolamine and NN-dimethylethanolamine. 3. When the analogues were incubated with [(3)H]glycerol and hepatocytes, the appearance of (3)H in unnatural phospholipids indicated that they were incorporated, at least in part, via CDP-derivatives. The distribution of [(3)H]glycerol among molecular species of phospholipids containing 2-aminopropan-1-ol and 1-aminopropan-2-ol was the same as in phosphatidylethanolamine. In other phospholipid analogues the distribution of (3)H was more similar to that in phosphatidylcholine. 4. NN-Diethylethanolamine stimulated both the conversion of phosphatidylethanolamine into phosphatidylcholine and the incorporation of [Me-(14)C]methionine into phospholipids. Other N-alkyl- or NN-dialkyl-ethanolamines also stimulated [(14)C]methionine incorporation, but inhibited the conversion of phosphatidylethanolamine into phosphatidylcholine. This indicates that phosphatidyl-NN-diethylethanolamine is a poor methyl acceptor, in contrast with other N-alkylated phosphatidylethanolamines. 5. These results on the regulation of phospholipid metabolism in intact cells are discussed with respect to the possible control points. They also provide guidelines for future experiments on the manipulation of phospholipid polar-headgroup composition in primary cultures of hepatocytes.


Loss of haem from cytochrome P-450 caused by lipid peroxidation and 2-allyl-2-isoprophylacetamide. An abnormal pathway not involving production of carbon monoxide.

1. Microsomal preparations undergoing lipid peroxidation produce CO and lose haem from cytochrome P-450. 2. The amount of CO produced does not correlate with the amount of haem lost and, after pre-labelling of microsomal haem in its bridges with 5-amino[5-14C]laevulinate, the radioactivity lost from haem is not recorved as CO. 3. Similarly, when pre-labelled microsomal haem is destroyed by the action of 2-allyl-2-isopropylacetamide, no radioactivity is recovered as CO. In clear contrast, on degradation of haem by the haem oxygenase system, CO is produced in an amount equimolar to the haem lost. 4. It is concluded that (a) the CO produced during lipid peroxidation originates from a source different from haem and (b) the degradations of haem caused by lipid peroxidation and 2-allyl-2-isopropylacetamide do not involve to any significant extent evolution of the methene-bridge carbon of haem as CO.


Clearance of human N-acetyl-beta-hexosaminidases from rat circulation.

Human N-acetyl-beta-hexosaminidases As and P are probably sialylated since they are susceptible to neuraminidase attack. twhen infused into the circulation of a rat they are removed more slowly than the non-sialylated forms of the enzyme from tissue and urine.


Activity of covalently cross-linked cholera toxin with the adenylate cyclase of intact and lysed pigeon erythrocytes.

Reaction of cholera toxin with NN'-bis(carboximidomethyl)tartaramide dimethyl ester produced several cross-linked species that had subunit B (which binds to the cell surface) and peptides A1 (which activates adenylate cyclase) and A2 all covalently joined together. This cross-linded material had activity with pigeon erythrocytes that was comparable in all respects with that of native toxin. It activated the adenylate cyclase of whole cells, showing a characteristic lag phase, and this activation was increased if the cells had been preincubated with ganglioside GM1, but abolished if the protein had been preincubated with the ganglioside. It activated the enzyme in lysed cells more strongly and without the lag phase. These results show that the toxin is active even when peptide A1 cannot be released from the rest of the molecule.


Control of glucose metabolism in isolated acini of the lactating mammary gland of the rat. The ability of glycerol to mimic some of the effects of insulin.

Inhibition of glucose uptake by acetoacetate and relief of this inhibition by insulin found previously in slices of rat mammary gland [Williamson, McKeown & Ilic (1975) Biochem. J. 150. 145-152] was confirmed in acini, which represent a more homogeneous population of cells. Glycerol (1mM) behaved like insulin (50 minuits/ml) in its ability to relieve the inhibition of glucose (5 mM) utilization caused by acetoacetate (2 mM) in acini. Both glycerol and insulin reversed the increase in [citrate] and the decrease in [glycerol 3-phosphate] and the [lactate]/[pyruvate] ratio in the presence of acetoacetate. Lipogenesis from 3H2O, [3-14C] acetoacetate, [1-14C]- and [6-14C]-glucose was stimulated, whereas 14CO2 formation from [3-14C]acetoacetate was decreased. Neither insulin nor glycerol relieved the acetoacetate inhibition of glucose uptake when lipogenesis was inhibited by 5-(tetradecyloxy)-2-furoic acid. From measurements of [3-14C]acetoacetate incorporation into lipid in the various situations it is suggested that a cytosolic pathway for acetoacetate utilization may exist in rat mammary gland. In the absence of acetoacetate, glycerol inhibited glucose utilization by 60% and increased both [glycerol 3-phosphate] and the [lactate/[pyruvate] ratio. Possible ways in which glycerol may mimic the effects of insulin are discussed.


The mechanism of C-20 hydroxylation of alpha-ecdysone in the desert locust, Schistocerca gregaria.

1. The C-20 hydroxylation of alpha-ecdysone to produce beta-ecdysone was investigated in the desert locust, Schistocerca gregaria. 2. alpha-Ecdysone C-20 hydroxylase activity was located primarily in the fat-body and Malpighian tubules. The properties of the hydroxylation system from Malpighian tubules investigated further. 3. The enzyme system was mitochondrial, had a pH optimum of 6.5, an apparent Km of 12.5 micron and required O2 and NADPH. 4. The activity of the hydroxylation system showed developmental variation within the fifth instar, the maximum activity corresponding to the maximum tire of endogenous moulting hormone. The significance of these results is assessed in relation to the control of the endogenous titre of beta-ecdysone. 5. The mechanism of the hydroxylation system was investigated by using known inhibitors of hydroxylation reactions such as CO, metyrapone and cyanide. 6. The CO difference spectrum of the reduced mitochondrial preparation indicated the presence of cytochrome P-450 in the preparation. 7. It concluded that the alpha-ecdysone C-20 hydroxylase system is a cytochrome P-450-deendent mono-oxygenase.


The pathway of glutamate metabolism in rat brain mitochondria.

1. The pathway of glutamate metabolism in non-synaptic rat brain mitochondria was investigated by measuring glutamate, aspartate and ammonia concentrations and oxygen uptakes in mitochondria metabolizing glutamate or glutamine under various conditions. 2. Brain mitochondria metabolizing 10mm-glutamate in the absence of malate produce aspartate at 15nmol/min per mg of protein, but no detectable ammonia. If amino-oxyacetate is added, the aspartate production is decreased by 80% and ammonia production is now observed at a rate of 6.3nmol/min per mg of protein. 3. Brain mitochondria metabolizing glutamate at various concentrations (0-10mm) in the presence of 2.5mm-malate produce aspartate at rates that are almost stoicheiometric with glutamate disappearance, with no detectable ammonia production. In the presence of amino-oxyacetate, although the rate of aspartate production is decreased by 75%, ammonia production is only just detectable (0.3nmol/min per mg of protein). 4. Brain mitochondria metabolizing 10mm-glutamine and 2.5mm-malate in States 3 and 4 were studied by using glutamine as a source of intramitochondrial glutamate without the involvement of mitochondrial translocases. The ammonia production due to the oxidative deamination of glutamate produced from the glutamine was estimated as 1nmol/min per mg of protein in State 3 and 3nmol/min per mg of protein in State 4. 5. Brain mitochondria metabolizing 10mm-glutamine in the presence of 1mm-amino-oxyacetate under State-3 conditions in the presence or absence of 2.5mm-malate showed no detectable aspartate production. In both cases, however, over the first 5min, ammonia production from the oxidative deamination of glutamate was 21-27nmol/min per mg of protein, but then decreased to approx. 1-1.5nmol/min per mg. 6. It is concluded that the oxidative deamination of glutamate by glutamate dehydrogenase is not a major route of metabolism of glutamate from either exogenous or endogenous (glutamine) sources in rat brain mitochondria.


Amino acid transport in brush-border-membrane vesicles isolated from human small intestine.

Uptake of L-alanine and L-phenylalanine by purified bursh-border-membrane vesicles isolated from human small intestine was investigated by using a rapid-filtration technique. L-Alanine entered the same osmotically reactive space as D-glucose, indicating that transport into the vesicle rather than binding to the membranes was being observed. The uptake rate for L-alanine was higher in the presence of a Na+ gradient than in the presence of a K+ gradient. In the presence of a Na+ gradient, the lipophilic anion SCN- caused an increase in L-alanine transport, whereas the nearly impermeant SO42- anion decreased the uptake of L-alanine compared with its uptake in the presence of Cl-. The uptake of L-phenylalanine into the brush-border-membrane vesicle was also stimulated by Na+. The results indicate co-transport of Na+ and neutral amino acids inthe human intestinal brush-border membrane.


A study of the kinetics of the muscarinic effect on phosphatidylinositol and phosphatidic acid metabolism in rat brain synaptosomes.

The uptake of [32P]phosphate into phosphatidylinositol and phosphatidate was measured in synaptosomes incubated in Krebs-Ringer bicarbonate buffer, pH7.4. The apparent dissociation constants for acetylcholine and carbamoylcholine was estimated from the increase in 32P uptake caused by these agents. These apparent constants were similar for both phosphatidylinositol and phosphatidate and were 2.7 +/- 0.5 MICROmeter for acetylcholine and 12 +/- 2 micrometer for carbamoylcholine when Ca2+ concentration was 0.75 mM. Under the same conditions the inhibition of the carbamoylcholine-induced increase in 32P uptake, caused by atropine, is consistent with atropine being a competitive inhibitor, with an apparent inhibition constant of 0.35 +/- 0.05 micrometer. The apparent constants were dependent on the Ca2+ concentration, and were greater in 2.54 mM-Ca2+. The former values for the kinetic constants are similar to the muscarinic-receptor dissociation constant, which indicates that the binding of the agonist to the receptor may be rate-limiting in this series of reactions when the Ca2+ concentration is 0.75 mM.


Occurrence of 3-hydroxyretinol in the freshwater fish Bagarius bagarius and Wallago attu. Isolation and synthesis.

A new chromogen that absorbs maximally at 329 nm was frequently found in the liver oils of the freshwater fish Bagarius bagarius and Wallago attu. It was shown to be a diester of 3-hydroxyretinol. 3-Hydroxyretinol was found to be a metabolite of lutein during the biosynthesis of 3-dehydroretinol in a freshwater fish. A new method for the chemical synthesis of 3-hydroxyretinol from methyl 3-dehydroretinoate is described. The possible pathway for the conversion of lutein into 3-dehydroretinol in vivo is discussed.


Phospholipid requirement for 2-acetamidofluorene N-and ring-hydroxylation by hamster liver microsomal cytochrome P-450 enzyme system.

A phospholipid requirement of 2-acetamidofluorene N- and ring-hydroxylation was investigated with partially delipidated microsomal fraction from livers of 3-methylcholanthrene-pretreated hamsters. Butan-1-ol extraction of microsomal fraction removed 90% of the total lipid content without any appreciable effect on microsomal proteins. Such extracted microsomal fractions had much lower capacity to N- and ring-hydroxylate 2-acetamidofluorene: 25 and 44% of control respectively. Addition of butan-1-ol-extracted total lipid restored both oxidations to some extent, whereas addition of phosphatidylcholine fraction restored both oxidations almost completely. Addition of synthetic phospholipid, dilauroyl phosphatidylcholine, restored both oxidations to a large extent, whereas synthetic dipalmitoyl or distearoyl phosphatidylcholine was ineffective in restoring these oxidations.


Transmembrane migration ('flip-flop') of cholesterol in erythrocyte membranes.

After exchange with [14C]cholesterol-labelled plasma lipoproteins for 0.5-4h, erythrocytes were extracted with bile-salt solutions. The extracted cholesterol (mainly from the outside of the erythrocyte membrane) had the same specific radioactivity as the residual sterol. Thus cholesterol equilibrates rapidly (half-time less than 1 h) between the two sides of the membrane.


Stability of rat skin collagen during recovery from under-nutrition.

The growth of young rats was arrested for 6 weeks from 48 h after receiving an injection of L-[5-3H]proline. The 3H in the hydroxyproline of the newly synthesized skin collagen remained steady during under-nutrition and did not decrease during the subsequent recovery period. It was concluded that in this animal model the renewed growth did not induce degradation of the pre-existing collagen fibres.


Effect of Methylene Blue on glutamate and reduced glutathione of rabbit erythrocytes.

Methylene Blue injected into rabbits resulted in increased glutamate, reduced glutathione and pyrrolid-2-one-5-carboxylate in erythrocytes. The increase in reduced glutathione was probably due to increased glutamate changing the feedback control of glutathione on gamma-glutamylcysteine synthetase.


Affinity chromatography of thiol-containing purines and ribonucleic acid.

Evidence is presented for the specific interaction of 6-mercaptopurine with mercurated cellulose. Following from this, a new method is described for the affinity chromatography of thiol-containing molecules and of RNA containing incorporated 6-thioguanosine on columns of mercurated cellulose. This technique may find application in the study of RNA metabolism and gene expression.


Lysosomes and membrane recycling. A hypothesis.

A mechanism for intralysosomal membrane recycling is proposed. After invagination of the lysosomal membrane during autophagy, intralysosomal vesicles are formed. It is suggested that membrane can bleb out from these internal vesicles, probably in the form of very small vesicles, and return to the external lysosomal membrane by membrane fusion. This mechanism would conserve lysosomal membrane during autophay, and is analogous to current models of plasma-membrane recycling. Its relationship to turnover of lysosomal-membrane proteins and other proteins is discussed.


[Uterine adenomyosis (author's transl)].

Uterine adenomyosis is a common, almost banal, lesion, since it is found in one uterus out of three by careful histological examination. In a continuous series of 926 hysterectomies for various indications, 246 adenomyosis lesions were demonstrated. In only 65 cases, adenomyosis was the sole histological lesion explaining bleeding or pain, symptoms refractory to routine conservative treatment (haemostatic curettage and/or hormone treatment) and therefore justifying surgical removal of the uterus. The role of adenomyosis in the failure of conservative treatment for diffuse fibromatosis at around the age of the menopause must be remembered. Almost 50% of specimens of hysterectomy for fibroma show evidence of associated histological lesions of internal uterine endometriosis. Whilst the hysterographic diagnosis may be suspected in almost 2/3 of cases, direct appearances of adenomyosis are found in only one case in four.


Blood filter evaluation.

Massive blood transfusion and extracorporeal circulation result in bombardment of the small pulmonary arterioles with micro-aggregates which are mainly composed of cellular degradation products, damaged platelets and leukocytes, fibrin strands, portions of cellular membrane and protein precipitates. Such amorphous material can cause patients to die of respiratory insufficiency without underlying chest trauma. Increasing amounts of stored blood are transfused to patients and extracorporeal circulation has become one of the most useful techniques for surgeons. Unfortunately, these procedures are often accompanied by disturbing post-operative consequences. This is because micro-aggregates invade the capillary network of several organs, especially lungs, kidneys, brain and retina. It is why blood filtration has recently gained added interest because of widespread efforts to minimise the number or emboli which are either transfused or reinfused to the patient through the blood return line.


Recent experience with the Baylor left ventricular bypass pump.

The current Baylor left ventricular bypass pump is a pneumatically actuated diaphragm pump lined with segmented polyurethane. It is a swirl design in which blood moves in a circular motion within the pump. It has tilting disc inflow and outflow valves and the stroke volume is 60 ml. It is intended for periods of use up to fourteen days. Eighteen devices have been implanted in normal calves. The mean duration of pump operation was eleven days. The pump-canula junctions and regions of the valve seats have been sites of thrombus formation and are being redesigned to eliminate areas of flow separation and stasis.


Left ventricular failure in calves produced by supravalvular aortic stenosis.

A stable model of left ventricular failure is required to test left ventricular assist devices. Supravalvular aortic constriction will cause left atrial pressure to rise to new levels which are stable for several days. When left atrial pressure becomes unstable, an automatically actuated left ventricular assist device will increase its output and prevent pulmonary edema.


No-reflow phenomenon and acute myocardial ischemia. The need for further investigation.

It has been shown that localized myocardial ischemia is accompanied by microvascular changes which produce capillary obstruction when blood flow is restored. This so-called no-reflow phenomenon has been noted in the brain, kidney, dermis and, more recently, in the myocardium. Ultrastructural studies have pointed out the role of myocardial and endothelial cells swelling. It seems likely that such damage of the vascular bed may constitute the first irreversible change during ischemia and result in failure to obtain successful reperfusion of involved myocardium. It can be suggested on the basis of recent clinical and experimental observations, that this phenomenon may play a role in subendocardial necrosis associated with cardiac surgery, and in myocytolytic necrosis. In both of these conditions, the most salient features are the microcirculatory defect and the reperfusion injury following transient ischemia. Further investigations are needed to determine the basic alterations induced by no-reflow phenomenon and the value of various prophylactic and therapeutic measures.


Dose-response relationship in the migration inhibition test using peritoneal exudate cells and blood leucocytes of tuberculin sensitive guinea pigs.

A comparison of the direct migration inhibition using peritoneal exudate cells and peripheral blood leucocytes from the same tuberculin sensitive guinea pigs was performed. Three antigen concentrations: 3, 15, and 75 microgram of PPD per ml were used. Both type of cells provided similar results except at early incubation intervals when leucocytes, in the presence of lower doses of the antigen, displayed stronger inhibition than peritoneal cells. Thus, in our study, peripheral blood leucocytes are at least equivalent to peritoneal exudate cells in the migration inhibition test.


Effects of busulfan (BU) on hemopoiesis and immune reactivity in dogs.

Hematologic and immunosuppressive effects of BU were evaluated in 21 dogs. Twenty to 30 mg BU/kg produced severe but in most cases reversible myelotoxicity. All animals given 40 mg BU/kg died of marrow failure which was consistently reversed by the infusion of autologous marrow. Granulocytic progenitor cells (CFU-C) in the bone marrow of BU-treated dogs were severely depressed by day 7 after drug administration and in surviving animals showed only slow recovery to normal values. Cell-mediated immunity as determined by the in vitro reactivity of lymphocytes was normal after stimulation with T-cell mitogens (PHA, ConA), but impaired after stimulation with B-cell mitogens (PWM, SpA). Antibody formation against sheep red blood cells was not suppressed by BU-treatment.


Changes of P50 in hypoxaemia, hypercapnia and polycythaemia: multivariate analysis.

Multivariate analysis of P50 changes in hypoxia, hypercapnia and polycythaemia was performed in an heterogeneous group of forty three patients: hypoxic subjects with or without hypercapnia, with or without polycythaemia and polycythaemic subjects without hypoxia. A statistical analysis was undertaken using comparison of the means, study of the correlations, principal component analysis, multiple regression and correspondence analysis. In the patients studied, P50 changes were not wholly explained by those of 2-3 DPG and pH; PaCO2, per se, did not play an important part. Haemoglobin concentration and P50 value would represent an adaptative mechanism to hypoxia: when hypoxia is moderate (80 greater than PaO2 greater than or equal to 65 torr) and isolated, oxygen haemoglobin affinity decreases (P50 increases); when hypoxia is severe (PaO2 less than 65 torr) and combined with hypercapnia and disturbed acid-base equilibrium, P50 comes back to normal range but haemoglobin increases, restoring thus, the normal blood oxygen content.


Possible high dosage gentamycin therapy in infected neutropenic patients.

Serum gentamycin levels have been studied in 23 neutropenic infected patients after high doses (62 mg/m2 and 72 mg/m2) of the drug, we observed: a higher gentamicin level than with an usual dosage, levels under the ototoxic 10/ml level, no renal failure, and, no patient ototoxicity.


Normal active rosette-forming-cells in untreated patients with Hodgkin's disease.

The percentage of E-rosettes and active E-rosettes was determined in untreated patients with Hodgkin's disease. All patients had numbers of peripheral blood lymphocytes within the normal range (1,200-5,000 lymphocytes/cu mm). The mean percentage of E-rosettes was significantly lower in the patients (55 +/- 15.7) as compared to normal controls (63 +/- 6.7). No difference in the percentage of active E-rosettes was found (36.6 +/- 8.6 in controls versus 40.3 +/- 10.8 in patients).


Nabilone: a potent antiemetic cannabinol with minimal euphoria.

Nabilone is a cannabinol derivative which has potent central antiemetic effects in animals. We observed that the drug significantly reduced the nausea and vomiting induced by cancer chemotherapy in 10 of 13 patients who were refractory to conventional antiemetics. A dose-response effect was apparent. The drug was generally well-tolerated, although it also had sedative effects. Additionally, dizziness, decreased coordination and postural hypotension were observed in some patients. Euphoric effects of the agent were minimal at antiemetic dosage levels.


Effect of a sulfhydryl inhibitor on in vitro bone marrow colonies (CFU-c).

A dose-related increase in the number of in vitro colony-forming units. CFU-c, was observed in mouse bone marrow cell suspensions following the administration of the sulfhydryl inhibitor, sodium iodoacetate. No effect on CFU-s was observed at the dosages and the periods selected for examination. Direct exposure of marrow cells in vitro to various concentrations of iodoacetate did not influence colony formation.


Studies on erythroid-committed precursor cells in the polycythaemic mouse.

The erythropoietin responsiveness of mice maintained in a polycythaemic condition for 42 days by transfusion of syngeneic red blood cells (but otherwise untreated) remained unchanged throughout the whole of that time. Furthermore the cycling rate, as measured by 3H-thymidine killing, also remained unchanged. These results indicate that continuous production and amplification of erythropoietin-responsive cells continues for long periods in the absence of demand for mature erythrocytes. It has also been shown that the erythropoietin response in the experimentally-induced polycythaemic mouse can be transiently increased as a result of "priming" injections of EPO. This suggests that the size of the erythropoietin-responsive cell (ERC) population has been increased, presumably by inducing extra division in the pre-ERC during maturation. This has previously been shown to occur only under conditions of drug induced depletion of both CFU-S and of ERC.


Regulation of haemopoietic stem cell proliferation in long term bone marrow cultures.

The development of a suitable bone marrow derived adherent cell population appears to be essential for the prolonged maintenance of haemopoietic stem cells in vitro. When established adherent layers are inoculated with freshly isolated bone marrow cells, proliferation of stem cells (CFU-S) regularly occurs both in the adherent layer and amongst the non-adherent cells. Furthermore, CFU-S present within the adherent layer are able to regenerate both themselves and the "non-adherent" CFU-S. One day after re-feeding the cultures (by removal of half the growth medium and addition of fresh medium) both the "non-adherent" and the "adherent" CFU-S are in a high cycling state (greater than 40% kill with 3HTdR). This proportion decreases with time of re-feeding and 5-7 days later the majority of "adherent" and "non-adherent" CFU-S are in a low cycling state ( less than 10% 3HTdR kill). Following a further re-feeding, CFU-S again enter a high cycling state.


Description of a suicide technique in vitro for granulocytic stem cells (CFUc) by hydroxyurea on normal human bone marrow.

It has been possible, by studying 15 normal human bone marrow samples, to develop a technique for the suicide effect in vitro by hydroxyurea (H.U.), applied to human granulocytic stem cells (CFUc). This study suggests a contact time of medullary cells from 60 to 80 minutes in the presence of a hydroxyurea concentration 6 mM per 10(6) cells per ml in vitro. Preliminary results concerning the clusters are discussed.


A microanalytic study of particles transport across the alveoli: role of blood platelets.

Following intratracheal injection of particles of colloidal gold (30 nm) in the rat, particles in the blood platelets of the alveolar capillaries can rapidly be observed. The presence of the gold is confirmed by microanalysis. The role of this phenomenon in pulmonary clearance is discussed.


Postulated mode of action of metals on purified human ALA-dehydratase (EC 4-2-1-24).

The effects of twelve metals at various concentrations ranging from 10(-41 to 10(-7) M have been studied on delta-aminolaevulinic acid dehydratase 9000 fold (ALA-D), isolated and purified from human red cells. The results obtained are in very good agreement with those of many authors: zinc, a constitutive element of the enzyme, behaves as an activator at low concentration, and an inhibitor at higher concentrations. The same effect is noted with aluminium, cadmium, mercury and tin. The manganese has a poor inhibitory action, copper and lead are powerful inhibitors of the enzyme. The other metals studies have no noticiable effect on ALA-D. These results agree with the following hypothesis: according to their structure, metals would bind the enzyme in one or several allosteric sites, and induce an allosteric transposition to the active or inactive form of enzyme.


Phenotypic variation in red cell G-6PD deficiency in heterozygotes.

Erythrocyte G-6PD activity in 69 heterozygotes has a log-normal distribution, a fact which cannot be solely attributed to random X chromosome inactivation. Others factors - genetic and individual - are suggested by intrafamilial clusting and post-natal variability of phenotype.


Restoration of the bone marrow pluripotent stem cells in AKR mice after arabinosylcytosine treatment.

The hypothesis of repression of multipotent stem cells (CFU) by leukemic cells to explain their depletion, previously demonstrated in AKR leukemic mice, was tested. Using arabinosylcytosine to destroy leukemic cells, it was shown that the bone marrow CFU pool was acutely depressed between 2 h and 12 h after treatment. However, 5 to 7 days later, this pool was restored, surpassing the value of the bone marrow pool in normal mice. This seems to indicate that the CFU pool in leukemic mice is potentially capable of proliferating but is repressed by leukemic cells.


Radioimmunoassay of anabolic steroids: an evaluation of three antisera for the detection of anabolic steroids in biological fluids.

Recently developed radioimmunoassays (RIA) for the analysis of anabolic steroids and their metabolites in biological fluids were tested for cross-reactivity with other types of steroids. Results show that the degree of desirable cross-reactivity within the two classes or orally active anabolic steroids vary widely and that the antiserum for 19-Nortestosterone (the active principle of intramuscular preparations) has a very high degree of undesirable cross-reactivity with components of oral contraceptives. Single and multiple dose studies in human volunteers demonstrate that the detection level and degree of retrospectivity are likewise variable but that the test easily detects most anabolic steroids during treatment. At the present time, the combination of the three antisera for the assay of a sample appears to be a relatively rapid and economic method for screening large numbers of samples in situations where doping control of anabolic steroids is required. The importance of utilizing physico-chemical means for identification of RIA potential positives is emphasized.


Responses to sustained use of anabolic steroid.

Description is given of six body-builders who had been taking Methandrostenolone (up to 20 mg/day in intermittent courses for a year or more). At the time of examination there was no subjective disturbance of sexual function, but testosterone levels were low relative to laboratory standards and luteinizing hormone levels were also reduced - particularly in relation to testosterone concentrations. Abnormal liver function tests were seen in three of the six subjects, and one had mild diabetes with high serum cholesterol, triglycerides and uric acid. The weight gain of the group was not outstanding, and the only possible finding was a high haemoglobin and haematocrit in one of the six subjects.


Screening tests for syphilis. A comparison of the Treponema pallidum haemagglutination assay with two automated serological tests.

Two Treponema pallidum haemagglutination assay (TPHA) tests (Wellcome and Fuzizoki) were compared with the automated reagin test and the automated Reiter protein complement-fixation test in a routine screening system. The Wellcome TPHA was shown to lack sensitivity, and it is recommended that it should be used at a lower screening dilution. The Fuzizoki TPHA recorded a significant number of false positive and false negative results, and was therefore inferior to the fluorescent treponemal antibody absorption (FTA-ABS) test as a definitive test for syphilis. It is suggested that the TPHA be used in conjunction with a reagin test to replace some of the more inadequate screening tests currently in use. The automated screening tests were shown to have a similar sensitivity to the TPHA as well as several other advantages; they are recommended for testing large numbers of specimens.


The impact of urban community hospital surveillance for gonorrhoea on the infection rate and complications in the female. A progress report.

This study substantiates a previous report concerning the importance of the urban community hospital, particularly its emergency room, in the detection of gonorrhoea in women. Pelvic inflammatory disease (PID), a major complication of gonorrhoea in women, significantly declined during a nine-year surveillance and control programme in the Memphis-Shelby County area. This suggests that the programme may have prevented PID developing in women through the early detection and treatment of asymptomatic infections.


The possible role of anaerobic bacteria in the aetiology of non-gonococcal urethritis in men.

Men with non-gonococcal urethritis (NGU) were divided into two groups and treated with either lymecycline or tinidazole; anaerobic cultures were performed before and after treatment. Neither treatment affected the anaerobic flora. However, the men treated with lymecycline were relieved of symptoms and signs, while those treated with tinidazole were not. The anaerobic bacteria found are considered normal inhabitants of the urethral tract and without significance in the aetiology of NGU.


Benign transient lymphangiectasis (sclerosing lymphangitis) of the penis.

The literature on benign transient lymphangiectasis of the penis (BTLP) was reviewed; reports of 45 cases were collected and a further series, comprising 21 cases, is presented. Both the published and present series support the view that BTLP is associated with recent sexual activity; little support is given for an infective cause and it is confirmed that BTLP is a benign, self-limiting condition presenting in the sexually active man between 20 and 40 years old. There can be recurrences, and susceptibility to them may be encouraged by circumcision in which disturbances of lymphatic drainage may play a part. As so few cases have been reported and few were diagnosed in the present series these comments can only be pointers to further studies. The descriptions in six histological examinations, the reports as `phlebitis' of apparently similar lesions, the report of `sclerosing lymphangitis' in other sites, and the known histological similarity between veins and lymphatics suggest that `sclerosing lymphangitis' is too specific a term. Until further knowledge is available, we suggest that this condition is best regarded as simple dilatation of a lymphatic vessel-namely, lymphangiectasis. As the condition is both benign and transient we propose the term benign transient lymphangiectasis of the penis for the condition formerly known as sclerosing lymphangitis.


A patient with primary syphilis of the hand.

The case history of a man with primary syphilis of the right hand is described. This type of luetic condition nowadays is rare. The incidence of genital, anorectal, and extragenital primary syphilis in both men and women and in sailors presenting at the Venereological Outpatient Clinic of the Department of Dermatology of the University Hospital Rotterdam-Dijkzigt during a period of seven years was studied. In sailors and women only genital primary lesions were found, whereas in male civilians 6.3% of cases of primary syphilis had anorectal lesions. The primary lesions in this study were different from those reported in earlier literature.


Unusual location of condyloma lata. A case report.

The case of a woman with secondary syphilis is described. The outstanding features were the presence of condylomata lata of the axilla and vulva as well as of the anal region.


The interrelationship between the olfactory bulbs and the basomedical hypothalamus in controlling food intake, obesity and endocrine functions in the chicken.

Surgical removal of the olfactory bulbs (OB) in the chicken indicated an increase in thyrotropic activity followed by a compensatory increase in food intake, as shown earlier. Basomedial hypothalamic (BMH) lesions in these birds caused changes in certain parameters which strongly infer hypoactivity of the thyrotropic axis. The lesions also caused development of typical hypothalamic obesity demonstrating both dynamic and static phases. Removal of the OB in BMH lesioned birds in their static phase of obesity, caused no demonstrable physiological changes. While removal of the OB caused an increase in somatotropic activity, the opposite was found in BMH lesioned birds. Those animals in which OB were removed and BMH was destroyed, demonstrated a decrease in somatotropic activity. It is suggested that the syndrome caused by BMH lesions dominates that caused by removal of the OB. The possibility of involvement of the OB in the control of thyrotropic and somatotropic activities mediated by the basomedial hypothalamus, is discussed.


Lightweight, multi-contact, slip-ring commutator for recording and stimulation with small animals.

A slip-ring commutator which can be used to electrically stimulate and/or record from the brains of small, unrestrained animals is described. In addition to providing 4 to 10 independent electrical contacts, featues of this model include its small size, low torque characteristics, low rate of contact oxidation, and minimal noise generated at the brush-ring surface. The compactness and light weight of this unit permit it to be suspended from a counterweighted boom assembly, thus providing additional freedom for vertical movement.


Infant rats: sensorimotor ontogeny and effects of substantia nigra destruction.

The ontogeny of sensorimotor behaviors of albino rats were evaluated from birth through adulthood (Experiment 1). Sensorimotor behaviors (e.g., visual and tactile orientation, forelimb and hindlimb hopping, righting reflexes) achieved mature (adultlike) characteristics at various ages during ontogeny and a rostral-caudal developmental pattern was revealed. In Experiment 2, the substantia nigra was bilaterally or unilaterally destroyed in rats at 10 or 25 days of age and the ontogeny of sensorimotor and regulatory (feeding, drinking, body weight regulation) behaviors were evaluated. Bilateral destruction of the substantia nigra, zona compacta, at 10 and 25 days of age resulted in transient cessation of suckling and/or feeding and drinking followed by recovery. Male brain-damaged rats had reduced body weight through 150-170 days of age. Specific feeding and drinking tests revealed the presence of residual regulatory deficits which seemed permanent. Sensorimotor testing revealed transient dysfunction for a variety of sensorimotor behaviors, with eventual recovery of normal sensorimotor capacity. The results are related to sensorimotor ontogeny and recovery from infant brain damage.


Ultrastructure of juvenile hormone-induced degenerating flight muscles in a bark beetle, Ips paraconfusus.

Topical application of 5 microgram of a juvenile hormone analogue (JHA), ZR-615, to female callow adults of Ips paraconfusus induced degeneration of the dorsoventral flight muscles. Within 24 h after JHA-treatment the diameter of the myofibrils was reduced to almost half due to the lysis of the peripheral myofilaments. Mitochondria showed conspicuous degenerative changes like swelling, dissolution of the matrix or presence in the matrix of dense filamentous material or myelin-like figures. Degeneration of the mitochondria seemed to take place inside isolation membranes derived from sarcoplasmic reticulum. A number of granular osmiophilic bodies appeared in the sarcoplasm. Three days after JHA-treatment the muscles were very thin and sheath-like. Most of the mitochondria had already degenerated. The dense sarcoplasm contained numerous crystalline bodies. The granular dense bodies were also more frequent. The myofibrils were comprised of only occasional small bundles of myofilaments. The tubules of the T system enclosed an amorphous material. The nuclei and the tracheal system remained intact but they were crowded due to the decreased volume of the muscle. In some specimens degeneration of the myofibrils and mitochondria was completed by the third day. No sign of degeneration was observed in the flight muscles of acetone treated control insects.


Neuronal and synaptic organization in the gravity receptor system of the statocyst of Octopus vulgaris.

The neuronal and synaptic organization of the sensory epithelium (macula) of the gravity receptor system of Octopus vulgaris was investigated by serial electron microscopic reconstruction. Three different types of afferent neurons, unipolar, bipolar, and multipolar, are described. Afferent synapses exist between the secondary sensory cells (hair cells) and the afferent neurons. Consequently, the neurons are first-order neurons. Two morphologically distinct types of afferent synapses could be identified: the most common type, present on every hair cell, has a finger-like postsynaptic process; the second type, which does not occur on every hair cell, has a flat or somewhat curved postsynaptic process. As a rule, the hair cells each form synapses with more than one afferent neuron. The neurons, in turn, form synapses with more than one hair cell. A complicated arrangement of efferent synapses was found at the level of both the hair cells and the neurons. The results are discussed with reference to their physiological consequences.


The retina-lamina projection in the crab Leptograpsus variegatus.

In the crab, Leptograpsus variegatus, the projection of retinula cell axons to the lamina was investigated by tracing them through a series of semi-thin sections. Forty-four such axons were traced from a single group of ommatidia as far as the distal layers of the lamina. The eight receptor axons of one ommatidium project to a single lamina cartridge. Therefore, because the crab has a fused rhabdom, angular information is conserved in vision, and the outside world is projected literally onto the lamina, just as it is in the standard non-dipteran pattern of insects. The belief of previous workers that other decapod eyes show neural superposition was an inference based primarily on the patterns of penetration of the basement membrane by receptor axons, and on degeneration experiments. This evidence is reviewed, shown to be inadequate and discussed in the light of the projection now demonstrated for Leptograpsus.


Freeze fracture studies on the annelid septate junction.

Freeze-fractured preparations of septate junctions between epidermal cells of annelids (Lumbricus terrestris and Tubifex spec.) have been investigated. In Lumbricus the protoplasmic face (PF) of the plasma membrane is characterized by variously arranged rows of particles. Apically the rows take an undulating course and often are separated by wide distances. In the basal part of the junction the rows run closely together and more or less in parallel. The diameter of the particles measures 80--120 A, the distance between two particles (centre to centre) is 150--250 A. Additionally striking rows of large particles (long diameter 150--200 A). Are to be observed mainly near the basal part of the junction. In Tubifex both faces of the plasma membrane could be studied in detail. The protoplasmic face (PF) contains rows of distinct individual particles (mean diameter 100--150 A, centre to centre distance approx. 250 A) whereas the particles of the extracellular face (EF, mean diameter 200-250 A) usually form continuous strands in which the individual particles seem to fuse. The density of arrangement of the strands varies considerably. Additionally ladder-shaped membrane structures have been observed in plasma membranes of this species.


"Sunbursts" and "christiesomes": cellular fragments in normal cow and goat milk.

Goats' milk includes numerous cell fragments ("christiesomes") which originate from the mammary secretory cells, contain well preserved endoplasmic reticulum, mitochondria and lipid droplets, and are responsible for the considerable triglyceride synthesising capacity of fresh goat milk. Cows' milk shows a few such particles only after repeated oxytocin-aided milkings. Cows' milk does contain quite different particles which have a dense content with a few small vesicles and numerous microvillus-like protrusions on one side ("sunbursts"). These have not been found in goats milk. Cytoplasmic particles similar to sunbursts have been found on the surface of the mammary secretory epithelium. It is suggested that they are residues of dead cells.


Fine structure of the drum muscles of the piranha (serrasalminae, characidae).

The anterior and the posterior drum muscles of the piranha resemble each other in all essential fine structural aspects: myofibrils are slender; sarcomeres are short compared with those of other drum muscles; mitochondria, located in the periphery of the fibers, are numerous and show an irregular internal structure; and the sarcoplasmic reticulum is abundant. Triads appear at the level of the Z lines. The drum muscles have many motor endplates, which, however, lack the characteristic junctional-fold apparatus. No lipid substances could be demonstrated in these muscles. In the posterior drum muscle the fibers depart from their orderly longitudinal arrangement at irregular intervals.


The effect of 1alpha-hydroxyvitamin D3 in pre-dialysis renal bone disease.

Assessment of 18 azotaemic patients treated with long-term 1alpha-hydroxyvitamin D3 (1alpha-OHD3) confirms the generally favourable effect of this analogue of 1,25-dihydroxyvitamin D3 in azotaemic osteodystrophy. Growing children with radiological rickets respond very well as do adults showing mild hyperparathyroidism with or without osteomalacia. However, patients with severe 'pure' hyperparathyroidism and features of autonomy do not respond well and in such patients 1alpha-OHD3 alone should be avoided. Phosphate restriction and occasionally a sub-total parathyroidectomy may be indicated in these patients.


Prophylactic 1alpha-hydroxyvitamin D3 therapy in haemodialysis patients.

Twenty-seven patients starting regular haemodialysis were treated with a 1 microgram daily dose of 1alpha-hydroxyvitamin D3 and concurrent aluminium hydroxide therapy to prevent hyperphosphataemia. There was an increase in plasma calcium, but no significant improvement in plasma alkaline phosphatase activity or parathyroid hormone levels. Metastatic calcification progressed but was never a severe clinical problem. Quantitative bone histology showed a significant decrease in resorptive surfaces confirmed radiologically, but there was no significant decrease in forming surfaces. The expected increase in forming surfaces with length of dialysis was however prevented.


The effect of 1alpha-hydroxyvitamin D3 on calcium and mineral content of bone in renal osteodystrophy.

Ten patients with renal osteodystrophy were treated with 1alpha-hydroxyvitamin D3 for 18 months. The effect of treatment was measured by a number of different methods and bone mass assessed by mineral and neutron activation analysis. In the majority of patients, raised serum levels of parathyroid hormone and alkaline phosphatase, histological features and radiological changes returned to normal. These effects were inconsistent in patients with a raised serum calcium level before starting treatment. In general, in patients without evidence of severe hyperparathyroidism there was a reduction in the rate of fall or an increase in the mineral and calcium content of the bone.


Spinal calcium changes with 1alpha-hydroxyvitamin D3.

During treatment of renal osteodystrophy with 1alpha-hydroxyvitamin D3 in eleven patients, regional changes in the skeleton have been compared with long-term calcium balance as assessed by whole-body calcium. Radial bone changes did not correlated well with calcium balance, but spinal calcium changes were of a similar magnitude when changes were large. Bone alkaline phosphatase changes correlated well with changes in spinal calcium, but less well with changes in radial bone density.


The effect of dialysate calcium concentration on 1alpha-hydroxyvitamin D3 on skeletal calcium loss and hyperparathyroidism in haemodialysis patients.

The response of hyperparathyroidism and skeletal calcium loss in haemodialysis patients to treatment with 1alpha-hydroxyvitamin D3 and a dialysate calcium concentration of 1.375 mmol/l was compared with the response to treatment with a dialysate calcium concentration of 1.375 or 1.75 mmol/l alone over a 6 month period. In patients treated with 1alpha-hydroxyvitamin D3 there was a significant rise in plasma calcium associated with a significant fall in plasma alkaline phosphatase and plasma parathyroid hormone as well as resolution of sub-periosteal erosions. In these patients there was a significant rise in the calcium content of the forearm assessed by neutron activation analysis in comparison to patients treated with a dialysate calcium concentration of 1.75 or 1.375 mmol/l alone. In patients treated with a dialysate calcium concentration of 1.375 or 1.75 mmol/l alone there was no significant change in the plasma calcium, alkaline phosphatase or parathyroid hormone after 6 months and in these patients subperiosteal erosions either did not change or became worse. No significant difference in the response in these two groups was observed. This study indicates that treatment of haemodialysis patients with 1alpha-hydroxyvitamin D3 is significantly more effective than treatment with a dialysate calcium concentration of 1.375 or 1.75 mmol/l alone in preventing progression of hyperparathyroidism and skeletal calcium loss.


The effect of 1alpha-hydroxyvitamin D3 with and without oestrogens on calcium balance in post-menopausal women.

Balance studies were performed in thirty-three post-menopausal women (all but five having vertebral crush fractures or femoral neck fractures) in the basal state and on treatment with 1alpha-hydroxyvitamin D3 and/or oestrogenic hormones. The results suggest that the effectiveness of oestrogen therapy is limited by calcium malabsorption and the effectiveness of 1alpha-hydroxyvitamin D3 is limited by oestrogen deficiency. The best results were obtained with combined therapy to remedy what appears to be two distinct deficiencies. To minimize the risks of hypercalcaemia and the possible risks of hormone therapy, we suggest that the treatment of choice in post-menopausal osteoporosis may be 1alpha-hydroxyvitamin D3 1microgram daily and ethinyloestrodiol 25 microgram daily for 3 weeks in every 4. Patients on a low dietary intake of calcium should probably be given calcium supplements. With this regimen, it should not be necessary to screen patients initially for calcium malabsorption or oestrogen deficiency because the majority of patients present with a combination of the two factors.


Effect of 1alpha-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 on intestine and bone in glucocorticoid-treated patients.

The effect of 1alpha-hydroxyvitamin D3 (1alpha-OHD3) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) on the intestinal calcium absorption was studied in twenty patients with rheumatoid arthritis treated with prednisone at daily doses of 5--15 mg for 1/2--20 years. The fractional calcium absorption, measured before and after the treatment with the vitamin D compounds, increased in nineteen of the twenty patients. This was, however, accompanied by marked rises in the urinary calcium excretion. There was no correlation between the fractional calcium absorption and the duration of the prednisone treatment or the doses given.


1alpha-hydroxyvitamin D3 in the long-term management of hypoparathyroidism and pseudohypoparathyroidism.

Normocalcaemia was restored and maintained in eleven hypoparathyroid and two pseudohypoparathyroid patients treated for up to 36 months with 1alpha-hydroxyvitamin D3. It was found to be a potent compound, maintenance doses ranging from 4 microgram weekly to 2 microgram daily. Supplementary oral calcium was used in acutely and profoundly hypocalcaemic patients but was given to only two patients as part of long-term therapy. Hypercalcaemic episodes occurring during treatment were of short duration and could be controlled by withdrawal of medication alone.


The management of hypoparathyroidism with 1alpha-hydroxyvitamin D3.

A retrospective study of the comparative effects of vitamin D, dihydrotachysterol and 1alpha-hydroxyvitamin D3 was undertaken in twenty-eight patients with hypoparathyroidism. The vitamin D compounds restored plasma calcium to the normal range in most patients with comparable actions on the gut, bone and kidney. Although the vitamin D compounds had a direct action on kidney and bone in the absence of PTH, the major action in maintaining plasma calcium was on the gut. Plasma phosphate fell due to a reduction in renal tubular reabsorption. Dihydrotachysterol and 1alpha-hydroxyvitamin D3 had significant practical advantages over vitamin D in their rapid onset of action and their relatively short biological half-life.


1alpha-hydroxyvitamin D3 in primary hyperparathyroidism.

The surgical management of primary hyperparathyroidism with severe bone disease (and tertiary hyperparathyroidism) has frequently been complicated by severe and prolonged post-operative hypocalcaemia. In seven such cases a short-pre-operative course of 1alpha-hydroxyvitamin D3 has been found to abolish these adverse features virtually completely and even to diminish pre-operative symptoms where present. Exacerbation of hypercalcaemia has not occurred and this regimen is thus recommended as a routine measure.


Histopathology of renal osteodystrophy with particular reference to the effects of 1alpha-hydroxyvitamin D3 in patients treated by long-term haemodialysis.

(1) The bone histology of 233 non-dialysed and 276 haemodialysed patients with chronic renal failure is reviewed. In non-dialysed patients osteitis fibrosa occurred in 83.7% and osteomalacia in 23.6% of patients. Osteomalacia was not found in the absence of osteitis fibrosa. In haemodialysed patients there was a more variable bone histology, sometimes resembling non-dialysed bone disease, but in general with a greater incidence of osteomalacia, especially with increasing time on dialysis. In some patients there was a predominance of osteomalacia accompanied by no or only mild osteitis fibrosa and the serum alkaline phosphatase was normal. (2) The results of treating twenty-six haemodialysed patients with 1alpha-hydroxyvitamin D3 (1alpha-OHD3) are described. Patients with osteomalacia and minimal or no osteitis fibrosa and a normal serum alkaline phosphatase (Group I) in general failed to respond and it is suggested that 1,25-dihydroxyvitamin D3 deficiency is not the sole factor responsible for the osteomalacia in these patients. In contrast, 1alpha-OHD3 therapy was effective in improving osteitis fibrosa and osteomalacia in some patients with moderate to severe degrees of osteitis fibrosa and osteomalacia (Group IIa) and in improving osteitis fibrosa where this occurred alone (Group IIb).


Bone histomorphometry in patients with chronic renal failure: effect of 1alpha-hydroxyvitamin D3.

Morphometric and dynamic studies of iliac crest bone biopsies were carried out in twelve patients with chronic renal failure (six on chronic haemodialysis and six with renal insufficiency) before and after 6 months of 1alpha-hydroxyvitamin D3 treatment. Pre-treatment biopsies showed signs of osteomalacia and hyperparathyroidism. Biopsies taken during treatment showed changes towards normality, indicating a beneficial effect of 1alpha-hydroxyvitamin D3.


Factors influencing the response to 1alpha-hydroxyvitamin D3 in patients with renal bone disease.

Twenty-three patients with bone disease and chronic renal failure were treated for periods of 4--28 months with 1alpha-hydroxyvitamin D3 (1alpha-OHD3). Improvements in bone histology were consistently seen in patients with features both of osteitis fibrosa and osteomalacia but were not invariably observed in patients with osteitis fibrosa or osteomalacia alone (37 and 50% improved respectively). Several factors influencing the outcome of treatment were assessed on the basis of histological responses in bone. A low level of plasma calcium before treatment, rather than the dose of 1alpha-OHD3 tolerated, was the major detectable factor which favourably affected the histological outcome. Other factors examined, including initial plasma concentrations of phosphate, immunoreactive parathyroid hormone and alkaline phosphatase, and treatment with haemodialysis or dietary supplements of calcium did not apparently influence the response.


Correlation of clinical, biochemical and skeletal responses to 1alpha-hydroxyvitamin D3 in renal bone disease.

Thirty-five patients with bone disease and chronic renal failure (twenty-four on maintenance haemodialysis) were treated for 7--39 months with 1alpha-hydroxyvitamin D3, 2--2.5 microgram daily by mouth. Symptoms (bone pain and muscle weakness) and radiographic appearances improved and plasma alkaline phosphatase returned to normal in the majority of patients (87, 76 and 75% respectively). In contrast, histological appearances in bone improved in only 46% twenty-three patients from whom paired biopsies were available, and this change was not greatly different from that seen in a comparable group of untreated patients. Significant correlations were noted in individual patients between the changes in symptoms, X-rays, plasma alkaline phosphatase and immunoreactive parathyroid hormone and these, in turn, were related to histological changes in bone, although these latter changes were often small. It is concluded that 1alpha-hydroxyvitamin D3 is a useful new drug in the treatment of renal bone disease, but that the evaluation of the response depends critically on the method of assessment used.


Hyperparathyroidism in chronic renal failure.

The high circulating concentrations of immunoassayable parathyroid hormone observed in chronic renal failure are due to a number of factors. These include altered metabolism of the hormone and also end-organ unresponsiveness which may, indirectly, cause increased secretion of parathyroid hormone. The response of the overactive parathyroid glands to changes in plasma calcium and magnesium is variable and caution is needed in evaluating the suppressibility of parathyroid hormone secretion in acute studies. 1alpha-Hydroxylated derivatives of vitamin D can effectively suppress parathyroid gland overactivity. This effect may not necessarily be medicated through hypercalcaemia and vitamin D metabolites may act directly on the parathyroid glands.


Effect of 1alpha-hydroxyvitamin D3 on parathyroid function in patients with chronic renal failure.

The effect of 1alpha-hydroxyvitamin D3 (1alpha-OHD3) on parathyroid function was studied in sixteen patients (ten on haemodialysis, six with chronic renal insufficiency) during treatment for 6--8 months. During treatment, a decrease in serum parathyroid hormone (s-PTH) and an increase in serum calcium were seen. An inverse linear correlation was found between serum calcium and logarithmic values of s-PTH in fifteen of sixteen patients, but the slopes of the regression lines varied considerably. This indicates that the suppression of parathyroid function seen during treatment with 1alpha-OHD3 is mediated by a rise in the serum concentration of calcium. For normalization of s-PTH, the serum concentration of calcium had to be above normal range in several patients, indicating a defect in calcium-mediated regulation of the secretion of parathyroid hormone. This abnormality was seen especially in long-term haemodialysed patients.


Transfer factor in the attempted treatment of patients with HBsAg-positive chronic liver disease.

Six patients with hepatitis B surface antigen-positive (HBsAg-pos) chronic liver disease have been treated with transfer factor (TF) prepared from leucocytes of normal blood donors with no history of hepatitis, and with TF from subjects recently recovered from type B hepatitis. In three patients there were transient elevations of aspartate transaminase (AsT) after 'specific' TF, representing damage or destruction of hepatocytes, and in two of these patients there was coincidental complement consumption, suggesting that TF had stimulated production of antibody. In one other patient there was an increase in E-rosetting lymphocyte (ERL) concentration representing a change in T-lymphocyte reactivity. One of the two patients who had no measured response to TF had a primary liver cell carcinoma and was receiving prednisolone therapy. TF prepared from subjects who have recently recovered from type B hepatitis may have temporarily altered the immunological status of patients with HBsAg-pos chronic liver disease, but it did not have a beneficial therapeutic effect.


Regulation of Leishmania populations within the host. I. the variable course of Leishmania donovani infections in mice.

The course of infection with Leishmania donovani was followed in seven strains of laboratory mice with measurement of the liver parasite burdens over 20 weeks. The acute parasite population growth rate varied greatly between, but not within, strains. Four strains were relatively resistant with less than an eight-fold increase while the three acutely susceptible strains showed over an eight-fold increase in the first month. Thereafter, one initially susceptible strain showed a dramatic fall in parasite numbers with histological liver damage while another strain maintained an immense parasite load for up to 2 years involving mononuclear phagocytes throughout the body. The system provides a model for studying genetic control of resistance to intracellular infection and the range of responses is compared with human leprosy and cutaneous leishmaniasis.


Regulation of Leishmania populations within the host. II. genetic control of acute susceptibility of mice to Leishmania donovani infection.

The acute growth rates of Leishmania donovani populations in twenty-five inbred mouse strains fall into two distinct groups: the susceptible (S) and the resistant (R). Hybrids within either category resemble their parents in susceptibility. Hybrids between categories are moderately resistant. Back-crossing of F1 hybrids to R and S parents, and interbreeding the F1 generation, give susceptibility ratios consistent with single gene control of acute susceptibility to visceral leishmaniasis. The distribution of this character among inbred mouse strains does not correspond to any well-studied gene nor does it appear to be linked to the H2 histocompatibility locus.


Agarose microdroplet leucocyte migration technique for detection of cell-mediated hypersensitivity to PPD and renal carcinoma antigen.

The agarose microdroplet leucocyte migration technique is an in vitro technique for detection of cell-mediated hypersensitivity in man, using only 10-15% of the volume of blood required for other leucocyte migration techniques. The technique is described in detail and results are presented from migrations with leucocytes from hypernephroma patients and controls tested with hypernephroma extract and normal kidney extract, and from tuberculin-positive and tuberculin-negative persons tested with PPD.


A study of the efficacy of typhoid vaccine in inducing humoral and cell-mediated immune responses in human volunteers.

The nature of protective immunity against typhoid fever in man is not at present well understood. Work on animal models and earlier studies from this laboratory indicate an important protective role for cellular immunity. The present work attempts to study the efficacy of the conventional typhoid vaccine in inducing specific cellular and humoral immune responses. The study on fifty-eight new army recruits and thirty-one civilian volunteers showed adequate humoral responses after vaccination. However, vaccination failed to induce a significant cellular immune response. In addition, a transient suppression of cellular immunity was observed in the immediate post-vaccination period in ten subjects who possessed natural cellular immunity before vaccination. These findings indicate the need for improving the typhoid vaccine so that it will induce cellular immunity as well as a humoral response. It also points to the necessity for obtaining detailed knowledge of the post-vaccination anergy as it could be important in timing public health programmes.


Acute energy deprivation in man: effect on serum immunoglobulins antibody response, complement factors 3 and 4, acute phase reactants and interferon-producing capacity of blood lymphocytes.

The effects of 10 days of total energy deprivation on serum levels of immunoglobulins, antibodies acute phase reactants and on interferon production were evaluated in fourteen healthy, normal-weight males. A significant depression was noted of the serum levels of complement factor 3, haptoglobin and orosomucoid. The titres of mercaptoethanol-sensitive specific antibodies to flagellin were higher in the subjects inoculated at the end of the starvation period than in controls and those inoculated at the start of the period. The serum levels of IgG, IgM, IgA, IgE, alpha-1-antitrypsin and complement factor 4, and the interferon-producing capacity of blood lymphocytes, were not changed. Thus, 10 days of total energy deprivation depresses the serum levels of several acute phase reactants and re-feeding may enhance antibody production.


Antibody-mediated adherence of rat eosinophils to schistosomula of Schistoma mansoni in vitro.

Eosinophils from the peritoneal washings of normal rats adhered to live or formalin-fixed schistosomula of Schistosoma mansoni in vitro, in the presence of heat-inactivated serum from infected rats. Eosinophil adherence caused permeability changes in the schistosomula as revealed by 51Cr release and methylene blue uptake. The serum factor which mediated adherence resided in the 7S fraction after Sephadex G-200 gel filtration. Protein A from Staphylococcus aureus which binds specifically to the Fc piece of IgG inhibited adherence, thereby demonstrating that IgG was the antibody responsible for this reaction and that the Fc portion was the site of interaction between eosinophil and antibody; rat eosinophils were shown to possess Fc receptors. The antibody mediating adherence reached high titres in the sera of rats 5-8 weeks after exposure to 500 cercariae, but thereafter there was a gradual decline in titre. Surface membrane from adult S. mansoni inhibited adherence, indicating the presence of cross-reacting antigens in adult worms and schistosomula.


The activation of the C3b feedback cycle with human complement components. I. Through the classical pathway.

Reaction between the fourth, the oxidized second and the activated first components of human complement generated the stable enzyme C4oxy2 capable of cleaving the third component and depleting total complement in human serum. This enzyme was shown further to activate the C3b feedback cycle as shown by its ability to consume factor B in serum and the reduction in the extent of complement consumption in the presence of EDTA. OxyC2 on its own gave rise to C3 cleavage in normal human serum by a pathway needing classical pathway components. This unexpected finding suggests that there may be a 'C-1 tickover' in serum analogous to the 'C3b tickover'; the presence of oxyC2 allowing the 'capture' of the trivial amounts of C42 normally formed. In preliminary experiments in the rat, C4oxy2 was successfully formed in vivo, where it gave rise to cleavage of C3, consumption of C5, depletion of cobra venom factor cofactors and a biphasic change in the neutrophil count.


The activation of the C3b feedback cycle with human complement components. II. Using components of the alternative pathway.

The C3 convertase of the C3b feedback cycle was generated in the fluid phase from C3b and factors B and D. It was shown to be an effective decomplementing agent when reacted with human serum. Many of the effects observed were dependent on the operation of the feedback during the ongoing reaction. Oxidization of factor B with iodine did not significantly increase the potency or the stability of the enzyme. Treatment of C3b with antrypol inhibited the formation of the enzyme. In preliminary experiments in the rat, the convertase was successfully formed in vivo, accompanied by cleavage of C3, consumption of C5, C6 and C7, and a biphasic change in the circulating neutrophils.


The complications of intravenous Corynebacterium parvum infusion.

100 intravenous infusions of Corynebacterium parvum were given to thirty-six patients at a dose of 5 mg/m2. Fever and rigors occurred in all patients but these acute side effects were acceptable to all but two patients. Seventeen patients suffered a delayed fall in blood pressure, which was marked in eight of them but was generally well tolerated. One patient died 18 hr after infusion from extensive myocardial infarction. Herpes labialis complicated the first infusion in nine instances, which may reflect transient immunosuppression following immunotherapy.


Interdependence of in vitro responsiveness of cord and maternal blood lymphocytes to antigens from oral bacteria.

A group of thirty-five mothers and their babies at parturition were examined by the in vitro lymphocyte transformation test to determine sensitization by oral bacterial antigens, B-cell mitogens and dental plaque. Lymphocytes from babies of sensitized mothers with gingival or periodontal disease gave the highest frequency (70 and 63%) and magnitude (mean stimulation index of 3.4 and 3.3) of response in cultures stimulated by Actinomyces viscosus and Veillonella alcalescens. However, IgM antibodies to V. alcalescens antigen were absent from cord sera. With one exception, stimulation of lymphocytes from babies of unsensitized mothers with clinically healthy gingiva was not found with these antigens. The response of cord lymphocytes from mothers with gingival or periodontal disease to antigens from oral bacteria, as compared with the response of cord lymphocytes from mothers with clinically healthy gingiva, seemed specific, since a corresponding difference in response to unrelated antigen PPD was not found. The response of cord and maternal lymphocytes to B-cell mitogens was also  determined. Maternal lymphocytes responded in the following decreasing order of effectiveness: dextran sulphate, levan, lipopolysaccharide and dextran B1355; whereas cord lymphocytes were stimulated in the reverse order of effectiveness.


The high prevalence of HLA-B5 in Behçet's disease.

Sixteen of the nineteen (84%) patients with Behçet's disease in Turkey had HLA-B5; the prevalence of HLA-B5 among 150 controls was 27%.


Serum immunoglobulin levels in N. brasiliensis infection.

We have measured the levels of IgG1, IgG2a, IgG2c, IgA and IgM in the serum of normal rats, and at various times after infection or re-infection with Nippostrongylus brasiliensis, in order to compare these responses with previously measured IgE levels. The results we have to report are that while the levels of the various immunoglobulin classes or subclasses are elevated to a greater or lesser extent, the increments are modest by comparison with the great elevation of total IgE.


Brain reactivity of lymphocytotoxic antibodies in systemic lupus erythematosus with and without cerebral involvement.

A prospective clinical study of cold-reactive lymphocytotoxic antibodies in systemic lupus erythematosus has been completed. A highly significant association between serum lymphocytotoxicity and the development of cerebral manifestations was observed. While lymphocytotoxic antibodies from patients with cerebral lupus were absorbed by homogenates of human brain, those from patients who at no time had evidence of cerebral disease failed to cross-react with brain. It is suggested that subpopulations of lymphocytotoxic antibodies differ in their brain reactivity, and that one population may be causally related to the development of some of the features of cerebral lupus.


A study of the material inhibiting EAC-rosette formation in the sera of patients with nephropathies.

Significant levels of EAC-rosette inhibition compared to control subjects were found in the sera of patients with focal and segmental hyalinosis (FSH), membranoproliferative glomerulonephritis (MPGN) and extra-membranous glomerulonephritis (EGN). In patients with IgA disease, although some sera produced high levels of inhibition, the group as a whole did not differ significantly from the controls. Evidence was obtained suggesting that the rosette inhibitory activity was due to immune complexes (IC) bearing C3 rather than C3 fragments. Firstly, the inhibitory activity was precipitable by 4% PEG, a concentration which does not precipitate the C3 fragments. Secondly, the inhibitory activity was selectively removed from the PEG precipitates by an anti-human immunoglobulin G immunoabsorbent. Finally, since it had been suggested that in some instances an unknown serum factor could inhibit EAC-rosette formation and activation of the alternative pathway of complement, the latter was studied and found to be normal in all the sera studied. Taken together, these results suggest that the inhibition of EAC-rosette formation obtained with the sera of the patients studied was due to the presence in these sera of some material behaving as IC. No clear-cut association was, however, seen between rosette inhibition and the presence or absence of Ig or C3 deposits in the kidney.