Concurrent occurrence of elevated ketone body and depressed carnitine level in underfed guinea-pigs

Summary Food ingestion of male guinea-pigs was restricted to 10 g/day. Total carnitine content of liver of the underfed animals fell to 3.75 moles per liver (46.1% of control). Serum level of total carnitine was 35.0 nmoles/ml (64.4%) in the underfed animals while they developed high ketonemia, 384.2 nmoles/ml (555.2%). In restricted animals the carnitine levels also decreased in the muscles.     

Changes in intracellular pH and cell volume during the early phase of DMSO-induced differentiation of friend erythroleukemia cells

Summary Changes in intracellular pH and water volume were measured after treatment of Friend erythroleukemia cells with 1.5% DMSO. It was found that a continuous decrease in pH_i occurred, beginning 1 h after induction and a decline in pH_i of 0.18 was measured after 9 h. In addition a decline in cellular water volume, of 12% only 15 min after induction, and 23% after 9 h, was observed.11 December 1986Acknowlegments. This work was supported by the Deutsche Forschungsgemeinschaft.     

Exchange of tritium from randomly tritiated taurocholate by microbial bile salt oxidoreductases

Summary Tritium distribution on randomly labelled taurocholate (TC) was estimated at 28%, 4%, 1% and <0.5% on the hydrogens opposite the 3-, 7- and 12-OH groups and taurine moiety respectively. Anomalously,C. perfringens 3-hydroxysteroid dehydrogenase (3-HSDH) catalyzed tritium loss of 36% on formation of 7-, 12-dihydroxy-3-keto-5-cholanoate, implying additional losses of tritium at other sites by this enzyme.This work is supported by the National Cancer Institute.     

A facile one-step synthesis of cysteinyldopas using mushroom tyrosinase

Summary A convenient one-step procedure, based upon the tyrosinase co-oxidation of dopa and cysteine, is reported for the synthesis of 5-S-cysteinyldopa (I) in 74% yield. Secondary products of the reaction turned out to be 2-S-cysteinyldopa (II, 14%), 2,5-S, S-dicysteinyldopa (IV, 5%), and the hitherto unknown 6-S-cysteinyldopa (III, 1%).The generic term cysteinyldopa is proposed to designate the various cotechol amino-acids arising from addition of cysteine to dopaquinone.This work was supported in part by a grant from Consiglio Nazionale delle Ricerche, Roma.     

Axillary 5α-androst-16-en-3-one in men and women: Relationships with olfactory acuity to odorous 16-androstenes

Summary Axillary 5-androst-16-en-3-one (5-androstenone) levels were found to be significantly higher in men than in women but do not vary between left and right axillae, are not related to age, handedness or degree of hirsutism (in women) nor to anosmia to this steroid. In men (but not in women), levels are related linearly to axillary cholesterol concentrations but not to squalene. Olfactory thresholds for 5-androstenone varied widely, the lowest recorded being 0.2 ppb, but there was no difference in thresholds between men and women. Women (70%) found the smell repellant but anosmia did not differ greatly between men and women (9–20%). Anosmia to the smell of 5-androst-16-en-3-ol was most marked in women (90%) rather than in men (45%). Axillary 5-androstenone values were generally consistent with the musky or strong smells of male axillary extracts, compared with the sweet smell of those from female subjects.Supported by the Herbert Dunhill Trust.     

Passive transfer of resistance in rabbits infested with adultIxodes ricinus L.: Humoral factors influence feeding and egg laying

Summary Partial immunity against the bites of I. ricinus was transferred to normal rabbits by inoculating immune serum from resistant animals. Transferred humoral factors diminished the weight f the ticks' blood meal by 29% and increased the feeding period by about 1 day in comparison with ectoparasites engorged on controls. They provoked also the failure of egg laying by I. ricinus. Only 55% of ticks fed on treated rabbits laid eggs (94% on controls). The immunological state of immune serum donors or recipients was studied and the IgG and homocytotropic specific anti-I. ricinus antibodies were identified. The immediate hypersensitivity of rabbits' skin was also controlled.The present work was supported by the Swiss National Fund for Scientific Research, grant No. 3.558.75.     

Kinetics of chemo-attraction of polymorphonuclear leukocytes towards N-formyl peptide studied with a novel polycarbonate (Nuclepore) membrane in the Boyden chamber

Summary The motile responses of human polymorphonuclear leukocytes (PMN) to N-formyl-methionyl-leucyl-phenylalanine (FMLP) in the Boyden chamber using a new sparse-pore polycarbonate membrane (pores 3 m in diameter and occupying 0.1% of surface area) were compared with those demonstrated by using a standard polycarbonate (Nuclepore) filtration membrane (pores 3 m in diameter and occupying 5% of surface area). Motility of PMN in gradients of FMLP using the new membrane was not influenced by chemokinetic effects of the factor, and the background migration of the cells was minimal. However, motility of PMN in gradients of FMLP using the standard membrane was found to be influenced by chemokinetic effects of the chemotactic factor, and the background or control migration (in the absence of chemotactic factor) of the cells was substantial. Greater directional migration of PMN according to steepness of the gradient of chemotactic factor was demonstrated with the use of the new membrane. The new membrane may be of considerable value in the further study of the chemotactic responses of PMN.     

Tumor necrosis factor-α inhibits collagen synthesis in human and rat granulation tissue fibroblasts

The purpose of the study was to examine the effects of tumor necrosis factor- (TNF-) on collagen gene expression in rat and human granulation tissue fibroblast cultures. The cells were exposed to 0.1, 1, 10, or 100 ng/ml of TNF-, and the rate of collagen synthesis was measured as synthesis of protein-bound³H-hydroxyproline. Total cellular RNA was isolated from fibroblasts, and measurements of specific cellular RNAs from fibroblasts were performed by Northern blot hybridizations using³²P-labeled cDNA probes. In cultures of rat granulation tissue fibroblasts TNF- decreased³H-hydroxyproline production to about 75% of that in controls and it also decreased pro1(I) and pro1(III) collagen mRNA levels, maximally to 33% and 23% of the control levels, respectively. In cultures of human granulation tissue fibroblasts a similar inhibiting effect in the production of collagen was seen. TNF- decreased the production of³H-hydroxyproline to 56% of the control value with a dose of 100 ng/ml also having an inhibiting effect on pro1(I) collagen mRNA levels of up to 43% of the control level. However, no effect was seen on pro1(III) collagen mRNA levels.     

Physicochemical studies of indocyanine green (ICG): absorbance/concentration relationship,pH tolerance and assay precision in various solvents

Summary Indocyanine green (ICG) obeyed the Beer-Lambert law within the concentration range 1.25 g/ml–10.0g/ml in distilled water, methanol, dimethylformamide (DMF), 1.2-propanediol and aqueous buffers (pH 9.0), but only up to 7.5 g/ml in human bile and 0.5% human albumin, and only to 5.0 g/ml in human duodenal fluid. ICG was rapidly (<1 h) decomposed to a colorless derivative at pH<5 and >11, but remained relatively stable for 48 h at pH 8–10. ICG is an indicator and a weak acid with a pKa of 3.27. In bile stabilized with 25% methanol, the precision of the method (CV) is 5% and the accuracy is 106–127%.     

The determination of extracellular space using hemoglobin

Summary The ECS of guinea-pig atria and frog sartorii can be determined using hemoglobin. For guinea-pig atria an ECS of (32.2±2.6)% wet wt for frog sartorii an ECS of (12.4±1.0)% wet wt can be measured.This work was supported by the Deutsche Forschungsgemeinschaft SFB 114 Bionach.     

An electrophoretic investigation of the binding of 3-14C coumarin to rat serum proteins

Summary The binding of coumarin to serum proteins of the rat has been demonstrated. Of the total bound coumarin (37% of injected dose), 36% was bound to the slow and fast ₁ globulins, 11% to the post albumins, 10% to globulin and 9% to albumin.     

The effect of dietary fibre on85Sr and47Ca absorption in infant rats

Summary The retention of bone-seeking radioisotopes⁸⁵Sr and⁴⁷Ca was about 20% lower in the femur of 6-day-old infant rats artificially fed on milk, milk supplemented with calcium and phosphorus, and milk supplemented with strontium when 6% cellulose type dietary fibre was added to the diet.Acknowledgments. This work was supported by a research grant from the Scientific Research Council of the Socialist Republic of Croatia. The authors wish to thank Mrs M. Buben, E. Herak, and K. Pribi for their technical assistance.     

Effect of air ionization on blood serotonin in vitro

Summary The effect of negative and positive air ionisation on siliconized blood serotonin was studied in vitro. The experiments showed that within 10 min positive ionisation increased serotonin levels in total blood (+40%), plasma (+90%), erythrocytes (+50%) and thrombocytes (+200%). On the other hand, negative ionization (10 min) lowered the serotonin content of total blood (–30%), plasma (–42.5%), erythrocytes (–41.7%) and thrombocytes (–72.3%), thus confirming the Krueger Effect in vitro.This research was generously supported by a grant from the Joint Research Fund of the Hebrew University and Hadassah, and Dr. and Mrs.Leonard Appleby of Palmbeach, Florida.Acknowledgment: Our thanks are due to Mr.B. Shalita, Mrs.Zippora Adar and Mrs.Olga Fradkin for devoted technical assistance.     

Fatty acid profiles of major food sources of howler monkeys (Alouatta palliata) in the neotropics

Wild howler monkeys (Alouatta palliata) get most of their calories from carbohydrates (65%) and fats (18%) of native tropical plants, but little is known about their intake of individual fatty acids. The fatty acid composition of several natural food sources of howler monkeys collected in Panama was determined by gas-liquid chromatography. The predominant fatty acids were palmitic (30%), linoleic (23%), linoleic (23%), -linolenic (16%) and oleic (15%). Fatty acids with less than 16, and more than 18, carbon chains were uncommon (0–7%). Although total saturated fatty acids were high in some specific food sources (22–54% of total fatty acids and 8 energy %), most of the calories from fat in the animals' diets are derived from mono-and polyunsaturated fatty acids (9.75 energy %) All food sources had significant amounts of the -3 fatty acid, -linolenic acid (2.9 energy %). In terms of human diets, the howler monkey's fat consumption would not be considered atherogenic. Unless these animals show a particular adverse susceptibility to dietary fat, it is unlikely that their fat intake is the primary cause of the low, but significant, incidence of atherosclerosis that develops in these animals in the wild state.     

A novel method in enzyme immunoassay: Maleimide derivative of hapten for enzyme coupling

Summary Meta-maleimidobenzoyl derivative of L-thyroxine methyl ester (MBTM) was synthesized and coupled to -galactosidase at molar ratio of over 5 to 1. More than 97% of the enzyme was found to be labeled with MBTM. A thyroxine enzyme immunoassay was carried out with sensitivity in the 0–10 g/100 ml range.     

Inhibition of respiratory oxidation of reduced sulfur compounds by intact cells ofThiobacillus denitrificans (strain RT) grown on thiosulfate

Thiobacillus denitrificans strain RT, an obligate sulfur-oxidizing chemolithoautotroph, was grown under microaerophilic conditions with thiosulfate as the only energy source. The rates of tetrathionate, thiosulfate, elemental sulfur (S^o) and sulfite oxidation were measured respirometrically with an oxygen electrode, using actively growing cells. Cells oxidized thiosulfate, elemental sulfur (S^o) and sulfite, but not tetrathionate. The thiosulfateoxidizing activity and elemental sulfur-oxidizing activity (SOA) were almost totally inhibited by 50 M myxothiazol (>80%), an inhibitor of the quinone-cytochrome b region, and by 10 M of the uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP) (>82%). Sulfite-oxidizing activity was also significantly inhibited (>60%) by 50 M myxothiazol and 10 M CCCP. 1 mM KCN totally inhibited (>90%) all respiratory activities. This study confirms that a sulfur-oxidizing activity appears during microaerophilic growth ofThiobacillus denitrificans strain RT on thiosulfate. The SOA is linked to the respiratory chain, probably releasing electrons in the quinone-cytochrome b region.To whom correspondence should be addressed. Submitted by R. Bachofen.     

Persistence of added retinoids in organ culture media during induction of mucous metaplasia and glandular morphogenesis in hamster cheek pouches

Summary The retinoid concentration (determined colorimetrically) did not change significantly in retinyl acetatesupplemented (6 g/ml) Eagle's Minimal Essential Medium containing 10% fetal calf serum when stored at –20 or 4°C over 7 days. After the medium was incubated at 37°C for 48 h, 37–49% of the retinoid remained, whether or not tissue (neonatal Syrian hamster cheek pouch) was present, and irrespective of explant age. The normal retinoid level in the tissue was approximately 0.25 g per gram. Therefore, neonatal hamster cheek pouches, incubated in medium with the addition of 6 g of retinyl acetate per ml of medium and undergoing mucous metaplasia and some mucous gland morphogenesis, were continually being exposed to retinoid levels which, though gradually decreasing, remained well above their normal physiological level.     

Self-labeling of human polymorphonuclear leucocyte myeloperoxidase with125iodine

In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with¹²⁵Iodine: chloramine T, lactoperoxidase, and an original technique of self labeling based on the ability of the enzyme to oxidize and bind¹²⁵I in the presence of H₂O₂. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 Ci/gmg MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.     

The instability of dietary iodine supply over time in an affluent society

In the Bernese region, where goiter was formerly endemic, alimentary salt has been supplemented by increasing amounts of potassium iodide (KI): 5, 10, 20 mg KI/kg in 1922, 1965 and 1980 respectively. Ioduria rose from <30 g I/g creatinine in 1920 to >100g I/g creatinine in the 1980s. In 1992 ioduria was estimated in 55 healthy volunteers (group A and individual B) and 234 thyroid carcinoma patients after thyroidectomy: hypothyroid patients with (C) and without thyroid remnants (D) and euthyroid patients on T4^** substitution (E). The arithmetic mean iodine excretion of the healthy volunteers in group A and individual B was found to be 87±40 g I/g creatinine. This is insufficient according to the recommendations of the WHO. In all groups, the iodine excretion reached the recommended level only in some members: 24% (A, B), 19% (C), 38% (D) and 81% (E).It was thought in the 1980s that in a formerly iodine-deficient society, iodinated salt would continue to provide an adequate supply of iodine. However, iodine intake in this affluent society has proved to be unstable. This can be attributed to modifications of eating habits, which include a reduction of total salt consumption, combined with a growing consumption of manufactured food of cosmopolitan origin, prepared using salt containing little or no iodine.     

Disruption of mitochondrial function as the basis of the trypanocidal effect of trifluoperazine onTrypanosoma cruzi

Summary The tricyclic anti-calmodulin drug trifluoperazine (TFP) inhibited growth and motility of epimastigotes ofTrypanosoma cruzi, at concentrations lower than 100 M, and motility and infectivity of the bloodstream trypomastigote form at 200 M. Electron microscopy of TFP-treated epimastigotes showed that the major effect was at the mitochondrial level, with gross swelling and disorganization. The oligomycin-sensitive, mitochondrial ATPase was completely inhibited by 20 M TFP, and the same drug concentration caused a 60% decrease in intracellular ATP content. The results suggest that the trypanocidal effect of TFP may be related more to mitochondrial damage than to the well-known anticalmodulin effect of the drug.