[Hypothalamic mechanisms of regulating blood circulation in the coronary and brachial artery systems]. In anesthetized curarized cats effects of high frequency stimulation of anterior, ventromedial, and posterior hypothalamus on arterial blood pressure (ABP) and on perfusion pressure of the coronary and brachial arteries were studied. The stimulation induced vasoconstriction of coronary vessels and increased the ABP. In most experiments, especially in case of stimulation of anterior hypothalamus, simultaneously with increase of perfusion pressure of the coronary artery, a decrease of the brachial artery perfusion pressure was revealed. The mechanism of vasodilatation of the brachial artery seems to be related to a direct inhibition of the tonic activity of the vertebral branch of the ganglion stellatum. The descending influence of the hypothalamus in respect to the activity of the inferior cardiac nerve is mainly facilitatory. Mechanisms of neural control of the regional blood flow in coronary and brachial arteries are discussed.


[Analysis of mechanisms regulating the heart in extreme situations]. Parasympathetic influences on functional and structural condition of myocardium in the process of stress modelling and adrenaline action on the background of the blockade of M-cholinoreceptors, was studied in 124 white rats. The stress was caused by means of severe exteroceptive stimulation or immobilization. Adrenaline and stress conditions caused impairment of automatism, change of conduct rate in the heart muscle and of force of excitement, anisotropia of muscle fibers, and penetration into the heart muscle cells of fluorescine. The effects were accelerated by diminishing of parasympathetic influences. The immobilization stress increased parasympathetic cardiac reflexes.


[Cardiovascular reactions to a change in the composition of cerebrospinal fluid]. In cats and dogs, cardiovascular responses were studied during stimulation of afferents, intraspinal pathways, after changes in composition of interneuronal media, 3rd ventricle's and spinal fluid. The longer hemodynamic responses as compared with the reflex and humoral those, preservation in conditions of spinal shock, their increased markedness together with weakening of the reflex regulation, their interrelationship with the reflex effects, their differentiation associated with the type of affecting drugs and with morpho-functional features of the central mechanisms of regulation and, finally, their integration with the reflex and hemato-encephalic influences in determination of the final hemodynamic effect,--all these constituted the six revealed features. Possible controlling role of the fluid effects is discussed.


[Pharmacologic analysis of the cholinergic vasodilation in unanesthetized animals]. Cholinergic vasodilatation evoked by hypothalamic stimulation in cats could be elicited by stimulation of both the emotional and mator areas of the brain. Tranquilizers affected the cholinergic vasodilatation in different ways: reserpine decreased, catapresan increased, while diazepam had no effect on amplitude of the cholinergic vasodilatation. The latter seems to be closely connected with activity of skeletal muscles but not with emotional reactions. Alpha--adrenergic structures of the c. n. s. are the activating mechanism of the central integration of cholinergic vasodilatation.


[Blood and cardiac cholinesterase activity in rats of different sexes and ages during muscle loading and hypokinesia]. The activity of acetylcholinesterase (Ache) and butyrilcholinesterase (Bche) in the blood and the heart of 3 and 13-month old control male rats is considerably lower than in female rats. In old rats (25 months) no sex differences in the Ache and Bche were revealed in the heart. In 3 and 13-month old male and female rats, under conditions of muscular exercises, the Ache and Bche activity is lower, and in hypokinetic male rats--higher than that in respective control animals. In all the rats, irrespective of sex, age and motor conditions, the Ache and Bche activity tended to decrease, in direction from the sino-atrial node to the heart apex.


Severe erosive stomatitis and giant lymph node hyperplasia of retroperitoneum (Castleman's tumor). A severe, extensive, erosive stomatitis developed in a 33-year-old Japanese man who had had intraperitoneal giant lymph node hyperplasia (Castleman's tumor) of the hyaline-vascular type. Histologically, there was a dense lymphoid cell infiltration in the dermis with invasion of the mucosal epithelium. It was hypothesized that an autoaggressive attack of lymphoid cells against the epithelium of the oral mucosa played a role in the pathogenesis of this erosive stomatitis and it was suggested that there might be a correlation between the occurrence of stomatitis and the presence of Castleman's tumor.


Trichilemmal cysts. Ultrastructural similarities to the trichilemmal sac. Recently experienced cases of trichilemmal cysts were studied by light and electron microscopy. Although ultrastructural findings in the present case were basically consistent with those reported in the literature, they were in contrast to them in some important respects: (1) a few, oval and small-sized keratohyalin and a large number of membrane-coating granules were observed in the cytoplasm of keratinizing cells beneath the keratinized cells and (2) the junctional structure between the keratinized cells and the keratinizing cells beneath them showed interdigitation. From these findings, it is shown that trichilemmal cysts have close similarities to and might be derived from the trichilemmal sac surrounding the lower end of the catagen or telogen hair.


[Treatment of lichen planus mucosae with vitamin A acid derivatives]. 27 patients with lichen planus mucosae or erosive lichen planus mucosae were treated with two vitamin A acid derivatives, 13-cis-retinoic acid or an aroumatic retinoid. 20 patients were definitively improved or healed. The treatment was especially effective in erosive lesions of the oral mucous membrane. Side-effects of this treatment are pointed out.


Oral contraceptives and their influence on porphyrin concentrations in erythrocytes and urine. In 15 females who have never before taken oral contraceptives, the porphyrin concentration in erythrocytes and in urine were investigated. The laboratory assays were performed before and after being on the oral contraceptive Stediril during 5 months (2 women took Stediril during only 3 months). The mean PP concentration in the erythrocytes increased from 16.4 microgram (before) to 24.1 microgram/100 ml of erythrocytes after taking Stediril regularly for 5 months. A statistical evaluation with the Student's test showed that at the level of 2%, the t(exp) = 2.58 was larger than the theoretical t(0.02) = 2.47. The difference of the mean CP concentration in the erythrocytes before and after taking Stediril was not statistically significant at the level of 5%. The mean concentration of CP in urine increased from 119.2 to 137.1 microgram in 1,000 ml. This difference was, however, not statistically significant at the level of 5% when assayed with the same test. There was no increase in UP concentration in urine.


[Experience with the photochemotherapy of psoriasis in Lausanne (author's transl)]. The results of photochemotherapy of psoriasis with oral 8-methoxypsoralen and long-wave ultraviolet light reported here are comparable to those observed in other centers using the same method of treatment. An average of 17-20 treatments was required to obtain clearing; 99-100% clearing was obtained in nearly 90% of the treated patients.


Plasma insulin response to arginine stimulation in children and adolescents with constitutional short stature according to sex, age, sexual development and skinfold thickness. The plasma insulin response to a standard arginine stimulation test was studied in 247 children and adolescents (177 males and 70 females) with constitutional growth retardation as compared with the response obtained in a matched group of 42 normal subjects and a group of 57 obese subjects. When the insulin response was related to pubertal ratings in the children and adolescents with growth retardation, it was found that the mean fasting levels and mean peak response both rose with the progression of puberty. This tendency was more pronounced in the females. When the insulin response was related to the degree of adiposity as measured by the degree of subscapular skinfold thickness, it was found that the greater the adiposity, the greater was the mean response, with the females showing higher levels than the males in each group. Even when the response in the two sexes was compared in subjects of a similar age and skinfold thickness, the females had a higher mean peak insulin response than did the males.


[Idiopathic hemochromatosis linkage with the HLA system (author's transl)]. Fourteen selected families containing two or more subjects suffering from idiopathic hemochromatosis and 34 unrelated cases have been studied for their HLA markers. A 3 was present in 75% of the unrelated cases vs 26% in the normal population (p less than 10(-8)). The frequencies of B 7 (38% vs 19%) and B 14 (23% vs 9%) were also increased (p lessthan 0,05). Inevitably, in most cases both antigens in the B locus were associated with A 3. Seven of nine affected sib pairs shared both HLA haplotypes, while two shared only one. Significant association between HLA haplotypes and diseases segregation has been demonstrated in family studies. These facts are consistent with the recessive inheritance of a strongly A 3 linked "disease" gene responsible for abnormal iron stores in the heterozygote state. This hypothesis would account for 64% of our present cases. Most of discordances (26%) were females who are physiologically protected, or children under 17 who might later develop the disease. The remaining 10% of disordant cases could be explained by crossing-over between "disease" gene and HLA loci or by an heterogeneity of the disease. This provides a method for screening for high risk subjects and perhaps an opportunity for anticipatory prevention.


Health behaviour and its determinants among insulin-dependent diabetics. Results of the diabetes Warsaw study. Health behaviour of a group of insulin-dependent diabetics and some factors influencing it have been described and analyzed. Aspects of health behaviour measured were: adherence to diet, insulin injection technique, urine self-testing and the pattern of action adopted in response to hypo- and hyperglycemia. Data were collected in 1975, from 170 insulin-dependent diabetics, aged 18-41, with known diabetes of from 6 months to 12 years duration. A standard questionnaire included items on health behaviour, knowledge of the disease and other factors. Of the subjects studied only 40% stated that they adhered to diet prescription. Occasional insulin injections were missed by 15%. Half the patients restricted diet or increased insulin dose in hyperglycemia but only 18% regularly tested urine for sugar. An important determinant of health behaviour was the level of understanding of the disease, which was found to be low among 58%. The present health education system for diabetic patients in Poland is discussed.


Absence of true pancreatic glucagon but persistence of circulating pancreatic glucagon-like immunoreactivity after pancreatectomy in pigs. The possible existence of circulating pancreatic type glucagon (immunoreactive glucagon as measured with a highly specific antibody) of extrapancreatic origin was investigated in 20-25 kg pigs after pancreatectomy. In intact conscious animals intravenous arginine infusions stimulated glucagon as well as insulin secretion, while blood glucose remained unaffected. Two weeks after pancreatectomy, and 48 hours after insulin withdrawal, basal glucagon and glucose concentrations were elevated (from 22 +/- 3.7 to 55 +/- 9.5 pmol/1 and 5.8 +/- 0.4 to 16.2 +/- 2.0 mmol/l, respectively), (mean +/- SEM), while insulin concentrations were either undetectable or very low. After pancreatectomy, however, glucagon concentrations no longer increased during arginine infusion. Gut type glucagon levels were not affected by pancreatectomy, and did not change during arginine infusion. When examined by gel filtration, plasma from unoperated pigs contained two components of pancreatic type glucagon, one coeluting with the glucagon marker, the other eluting earlier, probably reflecting larger molecular size. After pancreatectomy only this larger component was found in the plasma. The role of this component in the control of blood glucose is unknown.


Partial lipodystrophy and insulin-resistant diabetes. The coexistence of partial lipodystrophy of unusual distribution (limbs, back and head) with insulin-resistant diabetes and severe retinal, renal, neurological and arterial complications is reported. The lipodystrophy followed juvenile arthritis (Still's disease) and the diabetes, initially asymptomatic, became insulin dependent and technically insulin resistant (200 - 300 units insulin/day). Severe hyperlipidaemia has been a feature of this syndrome, probably contributing to the conspicuous peripheral arterial disease. The mother was diabetic but three sisters had normal glucose tolerance and there is no lipodystrophic member of the family. Underlying mechanisms of this syndrome remain obscure.


[Adipose tissue cellularity and skinfold thickness of the deltoid and retrotrochanteric areas in obese women (author's transl)]. 301 women, 261 of them obese, were the subject of an anthropometric study of brachial and femoral adipomuscular ratio, adipocytometry in the deltoid and retro-trochanteric areas and oral glucose tolerance. Deltoid adipocyte volume was highly correlated (p less than 0,001) with total fat deposition in this area, while no such relationship was found in the retrotrochanteric area. In obese women with chemical or overt diabetes, deltoid adipose cells were significantly larger than those of non-diabetic obese women. This, however, was not the case for retro-trochanteric adipose cells.


Evaluation of oral glucose tolerance test results in pregnancy. Oral glucose tolerance and insulin response to glucose were analysed in 124 pregnant women during the fourth quartile of pregnancy. Employing different criteria for the detection of glucose intolerance, 9% to 21% of women were abnormal, and using the H index 43% would have been declared "diabetic". There was no evidence of a progressive change in the glucose curve detectable by the H index within the fourth quartile of pregnancy. There was no association between actual or potential fetal morbidity and any of the interpretative criteria employed. It is concluded that the oral glucose tolerance test should be interpreted with caution if non-pregnant criteria of abnormality are employed.


Interaction of arginine vasotocin and norepinephrine upon pineal indoleamine synthesis in vitro. The interaction of arginine vasotocin (AVT) and norepinephrine (NE) upon pineal gland indoleamine synthesis was investigated. Rat pineal glands were incubated for 10 h in Krebs--Ringer bicarbonate plus 2 mg/ml glucose, 1 mg/ml bovine serum albumin, [14C]tryptophan, NE (10(-6) M), and log doses of AVT ranging from 100 ng to 10 microgram. Incubation media were extracted for [14C]serotonin while the other [14C]indoleamines, melatonin, hydroxyindoleacetic acid (HIAA), methoxyindoleacetic acid (MIAA), N-acetylserotonin (NAS), hydroxytryptophol (HTOL), and methoxytryptophol (MTOH) were separated by thin-layer chromatography. Serotonin metabolism was decreased by 0.1 microgram AVT and NAS decreased by 1.0 microgram AVT. Melatonin synthesis was decreased by both 0.1 and 1.0 microgram AVT. AVT also decreased the conversion of [14C]serotonin to MIAA and to HTOL. The data indicates that AVT decreased NE-stimulated pineal indoleamine synthesis in vitro and further suggests that AVT may participate in the intracellular regulation of melatonin synthesis.


Effect of replacement of extracellular sodium ions and of D-600 on the activation by adrenalin of adenylate cyclase in intact pigeon erythrocytes. The effect of replacement of extracellular Na+ by Li+, choline K+ or sucrose on cyclic AMP formation in pigeon erythrocytes has been investigated. Replacement of extracellular Na+ by Li+, choline or sucrose but not by K+ inhibited the stimulation by adrenalin of cyclic AMP formation, but had no detectable effect on cyclic AMP content in the absence of adrenalin. This inhibition was observed in the presence or absence of extracellular Ca2+. The relative inhibition caused by Na+ removal decreased with increasing adrenalin concentration. It was concluded that extracellular Na+ or K+ ions were required for maximal activation of adenylate cyclase by low concentrations of adrenalin, and that this effect of monovalent cations may have been due to an effect on the affinity of the receptor for adrenalin. The verapamil derivative D-600 also inhibited the stimulation by adrenalin of cyclic AMP formation. This effect occurred in the absence of extracellular Ca2+ and hence seemed to be unrelated to the inhibition by C-600 of the slow Ca2+ channel in electrically excitable tissues.


Biological and immunological characterization of human luteinizing hormone: IV. Biological and immunological profile of two international reference preparations after electrofocusing. The first IRP of Human Pituitary Gonadotrophins (FSH/LH) for bioassay (69/104) and the 1st IRP of Human Pituitary Luteinizing Hormone (LH) for immunoassay (68/40) were fractionated by an electrofocusing technique in a sucrose density gradient and the profile of biological and immunological activities was determined. The partially purified LH preparation (69/104) gave a broad pattern of biolgical and immunological activities which extended from pH 4 to 10. The biological : immunological (B/I) ratio of the various fractions (using the 68/40 preparation as standard) ranged from 0.04 to 1.05. The low B/I ratios indicate the presence of high levels of immunologically active, biologically inactive material in this preparation. In contrast to the behaviour of the 69/104 preparation, the major proportion (88%) of the biological activity recovered after electrofocusing of the highly purified LH preparation (68/40) was found within the pH range 7--9, with B/I ratios (again using as standard the 68/40 preparation) ranging from 0.4 to 1.5. Multiple dose parallel line design radioimmunoassays revealed a lack of parallelism between the 69/104 and 68/40 International Reference Preparations. This was attributed to the presence of acidic material in the former preparation, which is practically absent from the latter. The biological LH profiles of both the 69/104 and 68/40 preparations differed from those previously reported for aqueous extracts of pituitaries. It is concluded that the dissimilarity in the electrofocusing profiles of the biological and immunological activities of the 69/104 preparation renders this preparation unsuitable as a reference preparation for the quantitation of biologically active LH by radioimmunoassay methods. Using the same criteria, the 68/40 preparation would appear to be a more suitable standard.


Studies on the binding of the estradiol-receptor complex to rat DNA fragments. The activation by estradiol of a series of reactions in hormone target tissues provides a model for the study of gene regulation in eukaryotic cells. We have examined the interaction of the estradiol--receptor complex with the various frequency classes of rat DNA to determine whether specificity can be detected at this level of chromosome organization. Interaction of the 8S estradiol--receptor complex with DNA was assayed by a change in the sedimentation rate of the complex. The receptor was found to bind to high molecular weight rat DNA as well as to 450 nucleotide long fragments of rat DNA. When the sheared DNA was separated by denaturation and reassociation into three frequency classes, the unique and moderately repeated sequences bound the receptor almost as effectively as the total sheared DNA. The resulting DNA--receptor complexes were sensitive to pancreatic DNAase. The highly repeated class of sequences bound the receptor less effectively than the other fractions. This did not result from the presence in this fraction of an inhibitor of binding, since addition of unique DNA fragments resulted in formation of a normal DNA--receptor complex. Nor was nuclease activity responsible since intact DNA could be isolated from the incubation mixture. An increase in length of the highly repeated DNA to 800 nucleotide long fragments caused them to bind the receptor almost as effectively as the other DNA fractions. These studies suggest that there may be sequences in eukaryotic DNA that preferentially bind the estradiol--receptor complex.


Chronic administration of pentagastrin: effect on gastric mucosal protein synthesis in rats. Groups of adult rats were injected with either saline or pentagastrin (500 microgramg/kg) for 14 days. The capacity of the gastric mucosal ribosomes to synthesize endogenous and exogenous mRNA-directed protein in a cell-free system was then investigated. Polyribosomal profiles were also analysed on sucrose gradients. The endogenous mRNA-directed incorporation of 14C-labelled amino acids into protein by the gastric mucosal ribosomes from the pentagrastrin-injected rats was found to be considerably greater than that of the control. In the presence of exogenous mRNA (poly-U) the polyribosomes as well as the endogenous mRNA-free ribosomes (run-off ribosomes) from the pentagastrin-treated group showed 86% and 136% increment in [14C]-phenylalanie incorporation as compared to the corresponding control preparation. The ribosomal ribonuclease activity between the groups was found to be the same. The results of the present investigation indicate that an enhancement in the translational capacity of the ribosomes is in part responsible for stimulation of gastric mucosal protein synthesis after chronic administration of pentagastrin.


High-affinity binding of the antiestrogen [3H]tamoxifen to the 8S estradiol receptor. The interaction of tamoxifen (ICI 46,474), a synthetic antiestrogen, with uterine cytosol proteins of immature calf and rat has been studied directly using the tritiated compound labeled with a high specific activity. The binding complexes were measured by the dextrancoated charcoal, protamine sulfate and hydroxyapatite assays. Scatchard plots revealed a single class of high-affinity (KD congruent to 1.7 nM) binding sites, with a binding capacity similar to that of estradiol. Competitive experiments showed the same binding specificity for estrogens and antiestrogens. Sucrose gradient analysis revealed an 8S binding protein which could be partially proteolysed by trypsin into a 4S binding protein. Kinetic studies showed that the association rate of tamoxifen was 5 times lower than that of estradiol and reacted according to a second order kinetics. The first-order kinetics of dissociation was considerably higher than that of estradiol, giving a half-dissociation time of 20--40 min at 0--2 degrees C. In some cases tamoxifen displayed two slopes of dissociation, but the proportion of the slow-dissociating complex was always inferior to that found with estradiol. In contrast to estradiol, the kinetic constants ratio (k-/k+) gave a calculated dissociation constant, similar to that determined in equilibrium conditions (KD), agreeing with a simple reactional scheme. We conclude that the antiestrogen tamoxifen binds directly to the 8S cytosol receptor for estrogens and not to another receptor for the antagonists. In contrast to estradiol, the antagonist is rapidly dissociated from the receptor sites and is unable to protect them against thermal inactivation. The affinity of tamoxifen for its receptor sites as determined directly is surprisingly high when compared to its affinity evaluated indirectly by competitive experiments. It is then suggested that the two ligands either bind on two different sites of the same protein or induce a different conformational change of the same binding site.


Accumulation of carnitine in rat epididymis after injection of [3H]butyrobetaine in vivo: quantitative aspects, and the effects of androgens and antiandrogens. After i.m. injection of [3H]butyrobetaine into intact and castrated rats, the specific activity of plasma carnitine remained nearly constant over 24--96 h and epididymal uptake of carnitine was constant per unit time up to 72 h. The uptake ratio of intact to castrated rats was high at 48, 72 and 96 h after injection. Administration of estradiol valerate over 20 days reduced carnitine uptake in epididymis. This reduction was dose-dependent when estrogen was administered i.m. at 0.33--10 microgram/day levels. A maximum reduction of 90% was obtained with the 10 microgram dose. A dose increase from 33 to 100 microgram/day caused no further reduction. Norspiroxenone (2--10 mg/day) and SK 7670 (1.5 and 7.5 mg/day) were less effective than estradiol valerate (10 microgram/day) in suppressing carnitine uptake in epididymis. Epididymal carnitine uptake in estradiol valerate treated rats (33 microgram/day for 20 days) increased in a time- and dose-dependent manner under testosterone propionate treatment (50, 250, 1250 microgram/day). Carnitine uptake increased to 80% of the nonsuppressed levels when testosterone propionate was adminsitered over a 6-day period at 1250 microgram/day. Dihydrotestosterone increased epididymal carnitine uptake to the same extent as testosterone propionate. delta4-androstene-3,17-dione and 5alpha-androstane-3alpha,17beta-diol (50 microgram/day) were less effective, stimulating uptake to only 15% and 40% respectively of the testosterone propionate (250 microgram/day) stimulated levels. Changes in epididymal carnitine uptake evoked by various experimental procedures were closely paralleled by weight changes in the ventral prostate. This response resemblance indicates a similarity between the androgen sensitivity of the prostate gland and that of the carnitine uptake system in epididymis. The dose-dependent effect of estrogen on the accumulation of epididymal carnitine, together with the marked responses induced in this system by manipulation of its androgen status, support a possible use for the system as an assay for androgen or antiandrogen potency in vivo.


Purification and characterization of a juvenile hormone binding protein from the hemolymph of the fourth instar tobacco hornworm, Manduca sexta. A protein which binds the insect juvenile hormone has been isolated from the hemolymph of the fourth instar tobacco hornworm, Manduca sexta (Lepidoptera). Bioassay and chemical characterization of the bound ligand from the purified binding protein indicates that this molecule is the primary macromolecule responsible for juvenile hormone transport in the hemolymph of this insect. The juvenile hormone binding protein has been purified using gel filtration, ion exchange chromatography and preparative polyacrylamide gel electrophoresis. The protein is a single polypeptide chain of about 28,000 daltons with a sedimentation coefficient of 2.2S and an isoelectric point of 5.0. Binding analysis using a hydroxyapatite batch assay indicates that the juvenile hormone binding protein has one binding site with a Ka of 1.2 times 10(7) M-1 at 4 degrees C.


[Preventive rabies vaccination using vaccine prepared from human diploid cells]. 228 persons have been previously vaccinated with inactivated human rabies vaccine Mérieux (HDC). They received three injections of 1 ml by the deep subcutaneous route with a one month interval. The vaccination is completed with a booster of 1 ml one year later. Serum samples for antibody titration are taken just at the time of each injection and also 21 days after the third injection and after the booster. Antibody titration is performed using two different methods: seroneutralization by intracerebral inoculation in mice and focus fluorescent inhibition test. Both methods give similar results. The antibody levels with this vaccine are very high and the average titers are about 1/511 at the day 28 (14,27 I.U./ml), 1/1.558 at the day 56 (59,15 I.U./ml), 1/2.783 at the day 77 (75,09 I.U./ml). In comparison with suckling mouse brain vaccine we obtain titres five times higher with the vaccine HDC (titer 21 days after complete vaccination). The titres are 1/1.109 for HDC vaccine and 1/225 for suckling mouse vaccine. The incidence of second reactions after the different injections is about 27%. They resulted essentially in light pain (21%), redness (17%) and light induration (10%) at the site of the injection during 24 to 48 hours. Fever was observed in less of 1% of cases and no one other general reaction was described.


Human rabies immunoprophylaxis using HDC (MRC-5) vaccine. In a field trial 181 high risk persons of both sexes between ages of 2--71 years were prophylactically vaccinated against rabies with the beta-propiolactone inactivated rabies-vaccine (hdc) mrc-5. 1 ml of vaccine was administered intramuscularly on days 0, 7 and 21. Serum samples were collected on days 0, 7, 21 and 35.  Neutralizing antibody titers against the rabies virus were determined in the rapid fluorescent focus inhibition test, 7 days after the first injection measurable titers of antibodies were found in all sera with one exception. On day 21 all persons showed a 100% seroconversion with a mean titer of 1:1000 (8,3 IU/ml). On day 35 the final mean titer measured 1:2400 (20 IU/ml). Major side effects during the prophylactic vaccination were not observed.


The continuing imperatives of diversity of rabies vaccine. Current preoccupation with and enthusiasm for the human diploid cell rabies vaccine are reminiscent of the advent of duck embryo vaccine (DEV) a quarter of a century ago. Development of the latter vaccine, considered both potent and entirely safe at the time, had the effect of paralyzing development of other vaccines, at least in the United States. Subsequent experience has demonstrated the DEV is neither potent nor entirely safe, although its use is not attended by the trepidation that accompanied the use of older vaccines. Availability in the USA of only one vaccine licensed for human use has created a hardship on both patients and physicians in as much as a large percentage of vaccinated persons suffer side effects of various degrees that pose problems for continuation of treatment. Experience has shown that most of those problems can be resolved by simple substitution of vaccine of another kind. Data obtained thus far indicate the marked superiority of diploid cell vaccine over DEV in several respects but it is not yet evident that it is completely free of allergen. Moreover, purified preparations of antigenic fractions have been obtained, and other vaccines have been shown to benefit, in apparent potency, from their capability of eliciting interferon formation. Economic considerations cannot be overlooked, and the advantage, in this respect, of suckling mouse brain vaccine in some areas of greatest need is evident.


Studies with human diploid cell strain rabies vaccine and human antirabies immunoglobulin in man. Seventy healthy veterinary students with no previous history of rabies vaccination were inoculated either with human rabies immune globulin (HRIG) alone, human diploid cell strain (HDCS) vaccine alone, or combinations of both. HRIG, prepared either at the Lister Institute or by Cutter Laboratories, was inoculated (20 i.u./kg) on day 0, vaccine was inoculated on days 0, 3, 7 and 14 either in 0.1 ml doses intradermally (i.d.) or in 1.0 ml doses intramuscularly (i.m.). Neutralizing antibodies were measured by the mouse neutralization test on samples taken from each group of 10 volunteers on days 0, 1, 2, 3, 7, 10, 14, 21, 30, 50 and 90. The treatments were well tolerated and no serious reactions were seen. Antibody estimations, as yet incomplete, indicate that in those subjects receiving HRIG alone, low levels of antibody were detectable in most but not all cases after 24 hours; peak titres (0.16 i.u./ml : 1 in 7.2) were observed on days 2 and 3 and became undetectable after 30 days. Most subjects receiving vaccine and HRIG also had detectable antibody at 24 hours and all had antibody by 48 hours. Peak titres (GM +/- 18 i.u./ml) were recorded at 21 to 30 days. The response to vaccine administered by either route does not appear so far to be markedly suppressed by HRIG.


Human rabies globulin and human diploid vaccine dose determinations. Veterinary students were injected with either human rabies immune globulin and human diploid rabies vaccine, or with human diploid vaccine alone. All those students who received three or more doses of the vaccine developed high levels of neutralizing antibodies by the rapid fluorescent focus inhibition test (RFFIT). The levels of early antibody in the students receiving the combination of globulin and vaccine depended very much on the dose of globulin. The neutralizing antibody levels in the sera of students who received 12 IU/kg were low 48 hours after the globulin injection: 6 of the 25 students receiving this dose were found to have no antibody, and eight were found to have titers between 1:6 and 1:10. When the globulin dose of 20 or 30 IU/kg was administered all students had early titers of 1:11 or above and all students subsequently developed high levels of antibody.


Serovaccination by human immune globulin and HDCS-vaccine against rabies. A serological study was performed in 16 volunteer subjects and 3 patients to determine the active antibody response after vaccination with HDCS vaccine alone and in conjunction with rabies immunglobulin of human origin (RIGH). When heterologous antiserum from horse or mule and any rabies vaccine is used for passive-active immunization, a pronounced interfering effect is observed, depending upon the relative concentrations of preformed rabies antibodies given and rabies virus antigen of the vaccines. When 20 IU/kg bwt of RIGH were applied simultaneously with vaccine on day 0 followed by a full series of post-exposure anti-rabies treatment, i.e. 6 doses of vaccine at days 0, 3, 7, 14, 30 and 90, no interference phenomenon was observed, as indicated by an additive effect between passive antibody and actively produced antibody. This was reflected by the observed increase of rabies antibody titre on day 7 post-vaccination and could be demonstrated in three different assays (MNT, CFT and HIT). From day 14 to day 90, no significant difference as to logarithmic increase, plateau formation and persistence of neutralizing and HI antibodies was observed between the serovaccinated and the control group. Serum samples obtained on day 455 from 12 individuals representing both study groups revealed comparable antibody values as assayed in the MNT or HIT respectively. Thus persistence and quantity of vaccine induced antibody to rabies virus, whether produced after plain vaccination or under sero-vaccination conditions, actually show no difference.


IgM and IgG antibody profiles after immunization of man with a HDCS-vaccine (preliminary communication). HDCS vaccine was given to 20 healthy, not rabies-exposed people on days 0, 3, 7, 14, 28 and 90. Ten of them received in addition 20 IU/kg of human antirabies immunoglobulin. Vaccine potency in NIH tests was 1.8 (WHO). Most individuals did not show any measurable neutralizing antibody titer on days 3 and 7. On days 14, 28, 90 and 300 the mean of titres was above 1:200; with one exception, no titer was lower than 1:50. The sera of days 14 and 90 were separated on "Sephadex G-200" columns. On day 14 at least 46% of the rabies antibody was found in the IgM fraction, while 15% was recovered from the IgG fraction. On day 90 the proportion recovered from the IgM fraction had decreased to 17%; the IgG antibody had increased to 30% of the titres found in whole serum. Almost no antirabies activity was found in the IgG fractions.


Serological investigations on rabies tissue culture vaccine (USSR) and development of short methods in post-exposure treatment. Comparisons have been carried out in the GDR between the Hempt-vaccine, which has been used until now, and a vaccine prepared in the USSR from the Vnukovo-32 strain on tissue culture. A trial of 1,800 treatments undertaken in 18 vaccination centres has clearly shown that the tissue culture vaccine is superior to the Hempt-vaccine and allows the number of injections to be reduced. The authors recommend three types of treatment--with or without the combination of antirabies immunoglobulin--according to the risk of exposure or the injuries sustained.


Humoral immunity conferred by the HDCS rabies vaccine Mérieux administered after exposure: 151 cases with and without specific immune globulin of human origin. Preliminary communication. The results of the analysis of the neutralizing antibodies measured at days 30 and 90 after vaccination are described in brief. Different lots of Mérieux vaccine were used, of which two were checked separately for relative antigenic value by the Federal Public Health Service (CH). The lowest antigenic value (lot 0420 M) was 1.6 in relation to the standard NIH vaccine No.182. The inhibiting effect of immune globulin was shown to be significant with this lot, but not with the other, more powerful lots. The attention is drawn to isolated cases which did not present antibodies--or not a sufficient count--on days 30 or 90 according to the RFFI test.


Results of rabies post-exposure treatment with antirabies serum and the human diploid cell vaccine in Iran. Confirmation of safety and antigenicity of HDCV vaccine in volunteers and in post-exposure treatment of severely injured patients. In some cases the use of antirabies serum inhibited, temporarily, the development of the active response to vaccine. During the last year about 2000 treatments were carried out : no failure nor undesirable reactions were observed.


Production of inactivated rabies vaccine for human use on WI38 diploid cells. Results of potency tests. Stability of the vaccine in liquid and freeze-dried forms. The purpose of the article is to give the experience gained with the manufacture of the vaccine for five years and to dicuss the potency results obtained. Sixty ampoules of WI38 cells were used. The processing of twelve of them was stopped for accidental reasons. Forty-eight lots were freeze-dried, four were rejected as final product, three lots because of low potency, one following some febrile reactions in children due to an unexpectedly high content in endotoxin. The activity level of each lot was determined: (1) by titration of the viral suspension before concentration in young mice by the intracerebral route or by the plaquing technique in monolayers of BHK 21 cells. The mean titer in mice is 10(6.5) per ml; on cells the titer is lower, mean 10(5.8) per ml, but more homogeneous, and (2) by the NIH potency test in mice: 90% of the lots have an antigenic value compared to the International Reference Vaccine above 2. There is not a good agreement between the results of the NIH potency test and the titer of the viral suspension. The antigenic values obtained on the freeze-dried vaccine after storage one month at 37 degrees C are the same as those for the vaccine stored at + 4 degrees C. The activity is not modified after storage for two years at + 4 degrees C in the freeze-dried or in the liquid state. The main drawback of the vaccine is its high cost. Some suggestions are proposed to try to lower it and to find a sound compromise between high quality and price.


Potency testing of diploid-cell rabies vaccine and correlation with immunogenicity. Two procedures for evaluating the potency of diploid-cell rabies vaccine (WRV) were compared by mouse-serum neutralization (MNT) and RFFIT serological determinations, using NIH Reference Rabies Vaccine No. 182 as base. Mean antibody-induction potency value ratios in female CD1 mice for 12 WRV assays were 5.12 by MSN and 4.57 by RFFIT compared with a 2.84 Antigen Value by the standard 28-day NIH mouse neutralization tests on the same lots. The mean antibody-induction potency values for 12 control DEV preparations were 0.71 (MSN) and 0.81 (RFFIT) versus 0.51 Antigen Value by NIH test. The statistical parameters of all respective means were similarly compared. MRV potencies of the order of magnitude shown correlated well with superior human immunogenicity but not linearly over a ten-fold range of antigen values from about 1 to 10. MNT antibody responses were 15-30 I.U. per ml of serum to WRV in this range (3-dose immunization); corresponding overall RFFIT values were 8 to 15 I.U. per ml. Control immunizations with DEV preparations yielded immune responses approximately one order of magnitude lower under comparable conditions.


Standardization of methods used for testing the safety of tissue culture rabies vaccines. Tissue culture rabies vaccines are essentially new preparations intended for prophylaxis and treatment of rabies. However, the methods used for their testing are mostly the same as those used for rabies vaccines prepared from brain tissue. Therefore, it is extremely important to develop and standardize methods for testing tissue culture rabies vaccines. The main criteria for the evaluation of Vnukovo-32 production strain have been formulated. The strain has been characterized in pathomorphological studies; the degree and nature of lesions produced in animal brain tissue by various doses of antigen and different routes of inoculation have been determined. In order to ensure the specific innocuity of rabies vaccines inactivated preparations are produced. However, with the current methods of inactivation (treatment with beta-propiolactone or phenol, ultraviolet irradiation and others) some viral particles may occasionally remain active. Besides, reparative processes may possibly occur in utraviolet-irradiated viruses. The penetration of even sublethal doses of live virus into the body is highly undesirable, since under certain conditions the long-term harbouring of the live viral particle may result in the development of various pathological conditions. In order to reveal minimum amounts of live virus in the preparation a method of additional passages in a susceptible cell system using the fluorescent antibody technique has been developed. By this method it has been possible to detect the presence of live virus in 10 out of 51 lots of vaccine under test.


Rabies vaccine HDC: toxicological studies in laboratory animals. A purified rabies vaccine prepared in human diploid cell cultures (Wistar 38 adapted Pitman-Moore strain) was tested in several animal species with single and repeated administration. Acute toxicity studies were performed in mice, rats, and guinea pigs with s.c. or i.m. injections of at least 1 human dose/animal. The results gave no evidence of toxic effects. Studies on local compatibility were carried out in white rabbits (i.c. and s.c.) and albino guinea pigs (i.m.). During the observation period of 1 or 2 weeks after inoculation no adverse reactions were seen. Pyrogen tests in rabbits with 6 lots of vaccines showed that rabies vaccine is pyrogenfree. Long-term studies with 4 injections into rats and dogs and observation over a 4-month period gave no alterations of hematological parameters and of the urinalysis. The behaviour of the animals was not altered. Short- and long-term experiments in animals showed the innocuity of this type of rabies vaccine.


A purified human diploid cell rabies vaccine. Continuous-flow isopycnic banding of rabies virus in sucrose gradient was used to purify the antigen for the preparation of a human diploid cell rabies vaccine. Prior to the addition of the stabilizer, the average specific potency of several vaccine batches was 234 antigenic values per 1.0 mg of protein. Further data regarding potency and stability of this vaccine are presented.


Cell-mediated immunity and postexposure protection from rabies by inactivated vaccines of tissue culture origin. A strong cell-mediated cytotoxic response (CMC), which was maximal 6-7 days after treatment, was generated in mice exposed to inactivated rabies virus vaccine. CMC response was linear and was a function of antigenic mass. Live attenuated viruses also generated a strong CMC response after intracerebral infection. Mice lethally infected with virulent strains of virus failed to develop CMC. There was, however, no difference in the rate and amount of virus-neutralizing or lytic antibodies and interferon induction in mice infected with virulent as compared to those infected with attenuated strains of virus. The level of T-cell effector function was found to be directly correlated with survival. Secondary CMC response to inactivated virus vaccine is strong, appears faster (3 days)and requires a smaller amount of antigen than does the primary CMC response. A consistent high level of CMC can be maintained for extended periods of time by repeated vaccine inoculations. When mice were treated with vaccine after being infected with street virus, high levels of CMC were found in effectively protected animals, but no CMC was found in unprotected animals. High levels of virus-neutralizing antibody were present in both surviving and dying mice. CMC responses to different antigenic components of rabies virus were also investigated.


Antigenicity of low concentrated HDCS vaccine with and without adjuvant as compared to the standard fluid formulation. It can be shown that 0.1% aluminum hydroxide is able to compensate a 90% difference in antigen content of a human diploid cell strain rabies vaccine in man. This conclusion, however, is drawn only from the formation of neutralizing antibodies (kinetics and antibody concentration). Further experiments should be performed to compare also the protective capacity of HDCS vaccines varying in antigen content and adjuvant. A sole reduction of viral antigen is reflected by a corresponding reduction in antibody formation provided the different doses fall within the logarithmic part of the dose-response curve.


Development and clinical trials of the new human rabies vaccine of tissue culture (human diploid cell) origin. A new type of rabies vaccine for human prophylaxis was developed by workers at the Wistar Institute, Philadelphia, Pa. Vaccine is prepared from rabies virus infected tissue cultures of human diploid cells and is inactivated following concentration and purification of virus. Vaccine is being successfully used for prophylactic and postexposure treatment in humans with reduced number of doses and is virtually free from adverse reactions associated with the use of other types of rabies vaccines.


Zonal centrifuge purification of human rabies vaccine obtained on bovine fetal kidney cells. Biological results. An inactivated human rabies vaccine prepared on bovine fetal kidney cells is concentrated and purified by zonal centrifugation. The peak of rabies particles is monitored by hemagglutination. Immunogenicity of the purified particles was evaluated by titration of specific antibodies from vaccinated animals. Protective activity of the vaccine was assayed on guinea pigs challenged with street rabies. Biological results were compared with those obtained with other tissue culture vaccines.


Structure and function of rabies virus glycoprotein. Of the three major proteins associated with the rabies virus membrane, only the glycoprotein (G-protein) was found to be located on the external surface of the viral membrane. A minor glycoprotein (gp 50) detected by SDS-polyacrylamide gel electrophoresis (PAGE) of rabies virus appeared to be a breakdown product of the G-protein. Glycoprotein prepared by treatment of rabies virus with Triton X100 and purified by isoelectric focusing was found to be homogeneous with respect to size and isoelectric point. The apparent molecular weight of this component isolated under nondenaturing conditions is approximately 400,000 daltons. The same material analyzed by PAGE was found to be comprised solely of polypeptide chains of the G-protein (MW 80,000 daltons). Therefore, the Triton X100-released material represents homopolymers of the G-protein. The purified rabies virus glycoprotein (G) is only structural protein of the virus that induces the formation of virus-neutralizing antibodies and which confers immunity to animals. The total protective activity of the virus was recovered in the purified G protein preparation. The protective activity of G protein increased with purification : 9 ng of G protein was required to protect 50% of the mice as compared to 1.63 microgram of the virus. The number of oligosaccharide side chains on rabies virus glycoprotein was investigated. Analysis of glycopeptides obtained by protease digestion of desialated glycoprotein revealed three discrete glycopeptides. Comparison of the protease digestion products from desialated and from untreated glycoprotein indicated a heterogeneity among the glycopeptides in the sialic acid content. Two major tryptic glycopeptides were isolated from desialated rabies virus glycoprotein and were analyzed after protease digestion; one contained two oligosaccharide side chains and the other contained a single oligosaccharide side chain.


Evaluation of the inactivated tissue culture rabies vaccine from the Vnukovo-32 strain. Results of its industrial production and field use for post-exposure immunization of man. During 1973-1977, 981,000 doses of rabies vaccine were prepared in primary young Syrian hamster kidney cell cultures infected with attenuated rabies virus (strain Vnukovo-32) and inactivated by UV-irradiation. Potency of the vaccine varied from 0.3 to 3.5. More the 53% of vaccine lots had potency in the range of 1.1-3.5 (NIH). Post-exposure immunization was carried out in 45,842 persons aged from 2 weeks to greater than 76 years. The vaccination course lasted from 2 to 25 days and was followed by 2 booster injections with the dose of 1.5-5 ml. The vaccine combined with gammaglobulin was injected into 2,856 persons; 19,764 persons had bites of dangerous localization (face, head, fingers). Post-exposure immunization was carried out in 4,416 humans who were bitten or licked with saliva of rabid animals (diagnosis was confirmed clinically or in the laboratory) and in 16,760 humans bitten by suspected animals (diagnosis remained unknown). None of those who received the complete course of antirabic treatment fell ill with hydrophobia. No neurological complications or other severe side effects were noted. Field trials and the results of studies of virus-neutralizing antibody titers in 390 treated people make it possible to conclude that tissue culture rabies vaccine from strain Vnukovo-32 is an effective preparation with low reactogenicity.


Delayed type hypersensitivity in mice immunized with inactivated rabies virus. Evidence of the DTH reaction is in agreement with in vitro observation done with the lymphoblast transformation test in rabbits (5,8) or with the cytotoxicity test in mice (7). The footpad test (FPT) fulfilled all criteria of DTH. A maximum swelling at the injection site was observed 24 h after challenge. FPT could not be transferred by immune serum. But both local and systemic transfer of dissociated spleen cells or lymph node cells sensitized recipient mice to the FPT. A lymphoproliferative response in the draining lymph node was observed after injection of RV, coinciding with the appearance of circulating cells mediating DTH and this response is not dependent upon a B-cell regulatoring mechanism (3). Evaluation of vaccines in vivo by specific DTH reaction is of interest for testing the cell-mediated immunity of these vaccines.


Some experiences with human diploid cell strain-(HDCS) rabies vaccine in pre- and post-exposure vaccinated humans. HDCS vaccine has been in official use in the F.R.G. for pre- and post-exposure vaccination of man, after having shown its superiority to the Hempt and duck embryo vaccine--as far as compatibility, antigenicity and protective capacity is concerned--since February 1977. HDCS vaccines of both manufacturers, Mérieux/Lyon and Behring/Marburg, did not produce any severe side effects in about 5000 vaccinees in West Germany when conditions of vaccine production were properly observed. The lack of neuropathogenicity of vaccines is stressed. Contamination by bacterial lipopolysaccharides, however, resulted in endotoxin shock in 32 out of 35 vaccinees with one vaccine lot. Testing for pyrogenicity of every vaccine lot should be carefully observed in future. When comparing antigenicity of different vaccine lots of both manufacturers, individual titers of complement-fixing and neutralizing antibodies were correlated with the antigenic values of the vaccines. Duration of immunity after one course of vaccination is approximately 2 years and can be extended by only 1 booster injection for an additional 3 years. Protective capacity of HDCS vaccine (using Essen scheme, 6 applications) seems to be very pronounced.  So far, between 1973-1977, 68 individuals under severe risk were vaccinated post-exposure in West Germany and 45 individuals in Iran from 1975-76. No cases of rabies occurred in the vaccinees. HDCS vaccine in man induces a very early antibody response with appearance of IgM rabies specific antibodies on day 3, and IgG rabies specific antibodies on day 7 and an early IgM/IgG antibody conversion at that time.


[Results of preventive rabies vaccination with a concentrated vaccine of the PM/WI38-1503-3M rabies strain cultured on human diploid cells. Preparation of mixed antirabies-antitetanus hyperimmune immunoglobulin by plasmapheresis of blood taken from vaccinated veterinary students]. Many thousands of people in France and abroad have already benefited from preventive rabies vaccination by means of a vaccine obtained from culture on human diploid cells, perfected ten years ago by R. Lang, the Institut Mérieux and the Wistar Institute. In addition to being well tolerated, the serological efficacy of this vaccine is such that 100% of the vaccines observed had a seroconversion after only two injections at an interval of one month. However, a booster dose should be given 6 to 12 months after the first injection, and a further booster 3 to 5 years later or on request in case of known contamination. These boosters, combined with an anti-tetanus booster, induce such high antibody titers--between 10-100 and even 1000 I.U./ml--that it is easy to obtain substantial batches of combined anti-rabies and anti-tetanus immunoglobulin from a small number of volunteers. The complete efficacy of this new vaccine reduces the number of systematic post-vaccinal serologic controls and its innocuity is such that an extended preventive vaccination programme may be carried out, for instance in the case of children living in areas known to be dangerous.


Bimodality of fasting and two-hour glucose tolerance distributions in a Micronesian population. While frequency distributions of glucose concentrations in Caucasian populations are unimodal, bimodality has been described in the Pima Indians, a population with an extremely high prevalence of diabetes. Venous plasma glucose concentrations at fasting and after a 75-gm. oral glucose load were determined in 596 Nauruans, a Micronesian population with a diabetes prevalence of the same order as the Pima Indians. In both sexes and in subjects 10 to 19 years, the frequency distributions of the logarithms of the fasting and two-hour glucose values were clearly unimodal. In most sex and age groups of 20 years and older, the frequency distributions of fasting and two-hour glucose values were bimodal and consistent with a model of two overlapping Gaussian distributions. This population is characterized by marked obesity. However, there was no significant difference in the degree of obesity between subjects in the first and second curves of the bimodal distribution. This makes it unlikely that the bimodality is a consequence of the marked obesity seen in both the Pima and Nauru populations. The data show that among Nauruans, as with the Pimas, the frequency distribution of glucose concentrations can be used to separate the population into normal and hyperglycemic groups.


Glycosylated hemoglobin in normal subjects and subjects with maturity-onset diabetes. Evidence for a saturable system in man. Concentrations of glycosylated hemoglobin (GHb) are elevated in diabetes mellitus and are believed to reflect previous metabolic control. To better define possible determinants of GHb in man, we investigated the relationship between GHb and both fasting plasma glucose (FPG) and basal insulin (IRI) in 42 normal subjects and 29 patients with maturity-onset diabetes. Concentrations of GHb in diabetic subjects (12.7 +/- 3.4, x +/- S.D., per cent total hemoglobin) were significantly higher than in normal subjects (8.2 +/- 1.2, p less than 0.001). In normal subjects, FPG (r = 0.52) and GHb (r = 0.58) (both p less than 0.001) correlated with age. GHb did not correlate with IRI in either group. However, GHb was closely associated with FPG in both normal (r = 0.60, p less than 0.001) and diabetic (r = 0.85, p less than 0.001) subjects. Linear regression analysis of the data for the two groups combined was highly significant (r = 0.91, p less than 0.001). However, the slope of the regression line for GHb versus FPG seen in normal subjects was significantly steeper than that of diabetic patients (p less than 0.005). A curve describing a nonenzymatic saturable model was also found to fit the data of the two groups combined (r = 0.85, p less than 0.001), suggesting the possible existence of a saturable system for glycosylation in man.


Obesity, hyperinsulinemia, and portal blood flow. A theoretic study of the effects of increased portal blood flow upon fasting peripheral insulin concentrations. A mathematical model of steady state insulin metabolism was developed to predict the effects of blood flow changes upon fasting arterial insulin concentration. In addition to blood flows to major organs and tissues, the model includes representations for insulin feedback regulation of hepatic insulin clearance and pancreatic insulin secretion. Within the range of insulin sensitivities assumed for hepatic and pancreatic feedback regulation and the range of initial insulin secretion rates employed, the model predicts that the fasting arterial insulin concentration rises by up to 20 muU. per milliliter as portal blood flow increases. The necessary elevations in portal blood flow lie well inside the range of increased total hepatic blood flow observed in obese subjects. The results of this study suggest that a significant portion of the elevation in fasting insulin concentrations seen in obesity may be the result of increased portal blood flow and that other clinical observations and metabolic changes that accompany obesity may also be related to altered portal flow.


Abnormal lactic dehydrogenase patterns in carcinoma of stomach and in peritumoral tissues. Segments of gastric mucosa from 32 surgical and biopsy specimens containing gastric carcinoma have been studied to determine the total content of lactate dehydrogenase (LDH). The LDH isoenzyme distribution patterns in tumoral tissues have been compared to that of peritumoral tissue and to the gastric mucosa of a healthy control group. LDH isoenzyme activity in the samples from tumoral tissue was significantly increased. The adjacent peritumoral tissue had a slight increment in the polypeptide M as compared to the distant and normal tissue. This suggests that LDH isoenzyme measurement may prove a valuable addition to histologic study, in the recognition of the tumoral distribution in stomach carcinoma.


Cellular and humoral indices of disease activity in inflammatory bowel disease. In 51 untreated cases of ulcerative colitis and Crohn's disease some cellular (neutrophil alkaline phosphatase activity, neutrophil NBT reducing capacity, and neutrophil and plasma lysozyme activities) and humoral (serum orosomucoid and serum haptoglobin) indices of disease activity were quantitated. The most pronounced signs of disease activity, thus, were found in severe cases of ulcerative colitis. Combining lysozyme activities with other disease activity indices seems to facilitate the distinction between severe cases of Crohn's disease and ulcerative colitis. Beyond this the addition of the humoral indices seemed not to offer substantial help.


Fecal chymotroypsin: a study on its diagnostic value by comparison with the secretin-cholecystokinin test. Fecal chymotrypsin (CT) activities were determined in 149 randomly collected fecal specimens of 80 patients in whom the pancreatic function had been tested by a secretin-cholecystokinin test. There was a significant correlation between fecal CT activities and outputs of trypsin (r = 0.3451, p less than 0.01) and amylase (r = 0.3285, p less than 0.01) in duodenal juice. Fecal CT activities were normal in all patients who--based upon enzyme outputs in duodenal juice after stimulation with secretin and CCK/PZ--were classified as 'borderline cases', in most patients with 'low-normal' pancreatic function, and in a significant number of patients with established insufficiency of exocrine pancreas. On the other hand, fecal CT activities were abnormal in patients with severely impaired output of trypsin in duodenal juice, and only 7% of the fecal specimens from patients with established normal function of exocrine pancreas had abnormal low CT activities. It is concluded that the sensitivity of the fecal enzyme method is rather low as compared to the secretin-cholecystokinin test, but that fecal CT determinations give valuable diagnostic information in patients with more pronounced insufficiency of the exocrine pancreas.


14C-D-galactose breath test for evaluation of liver function in patients with chronic liver disease. D-Galactose metabolism and demethylation of aminopyrine by healthy controls and patients with chronic active hepatitis (CAH) and cirrhosis (Ci), were assessed by a breath analysis technique measuring 14CO2 exhalation after oral ingestion of 14C-D-galactose or 14C-aminopyrine. Patients with CAH and Ci exhibited decreased 14CO2-exhalation rates following 14C-D-galactose or 14C-aminopyrine. D-Galactose oxidation capacity of the liver can be assessed by a breath analysis technique in analogy to the demethylating function for aminopyrine. The ordinary oral D-galactose tolerance test seems, however, superior in comparison to the 14C-D-galactose tolerance test, in discriminating between healthy controls and patients with chronic liver disease.


Perhexiline maleate-induced hepatitis.

Perhexiline maleate-induced hepatitis. 5 patients treated with perhexiline maleate, 200-400 mg/day for at least 6 months, exhibited evidence of hepatitis. The picture was very similar to acute alcoholic hepatitis, clinically, biologically and histologically with presence of necrosis, Mallory's hyaline, polynuclear infiltration and to a lesser degree, steatosis. Association with peripheral neuropathy, hypoglycemia, and renal failure appears strikingly frequently. The evolution was severe since 3 patients died within 6 months, even after treatment withdrawal. Further studies are to be done to understand the mechanisms of hepatic and neurologic toxicity, and to measure the hazards of this drug. These studies could bring a new insight to alcohol toxicity.


The mechanism of anterior-posterior polarity control in planarians. The substance which inhibits brain formation in the regenerating planarian Dugesia etrusca was found to be a large molecule, at least in part protein, which electrophoreses as an electronegative moiety in pH 6.8 buffer. A model is presented, based on this finding and previous studies, which proposes an electrochemical mechanism for the control of polarity and possibly for the maintenance of tissue organization in planarians. It is proposed that a bioelectric field exists and moves the electronegative brain-inhibiting substance in a posterior direction, establishing polarity. This model explains the polarity reversal experiments using external fields and many of the previously unexplained classical planarian experiments. Data are presented demonstrating the existence, magnitude, and polarity of this bioelectric field, which is not greatly altered upon decapitation, all in accord with predictions of the model.


[Hemodynamic classification of acute myocardial infarction: physiopathological aspects and prognostic implications. Considerations on 130 cases (author's transl)]. Hemodynamic evaluations of 130 patients with acute myocardial infarction were performed by right and/or left heart catheterization. 115 patients were subdivided in six groups by the pulmonary artery mean pressure (PMP)-left ventricular stroke work index (LVSWI) relationship: 1) normal LVSWI in relation to PMP (14.8% of all cases); 2) increased LVSWI in relation to PMP (0.9%); 3) moderately reduced LVSWI with increased PMP (severe heart failure-cardiogenic shock) (7.0%); 5) reduced LVSWI with low or normal PMP (22.6%); 6) normal LVSWI with elevated PMP (reduced left ventricular compliance or high pulmonary vascular resistance) (15.7%). 28 cases were studied by right and left heart catheterization; in 10 cases only left heart catheterization was performed. Discriminant analysis on the values measured at the first stage of hemodynamic monitoring was conducted: this type of mathematical analysis seemed to provide a more useful prognostic index.


[Electrophysiological effects of ajmaline on the human conduction system (author's transl)]. The electrophysiological effects of ajmaline have been investigated using the intracardiac electrograms and the extrastimulus method. The drug was infused intravenously into 21 patients at the dose of 1 mg/Kg. Ajmaline induces:--variable effect on the sinusal cycle length--variable effect on the corrected sinus node recovery time--increase of the atrial effective refractory period (P less than 0.001)--in AV node slowing of conduction (P less than 0.001), a slight prolongation of PRE (P less 0.01) and more marked one of the FRP (P less than 0.001). The His-Purkinje (HV interval) and intraventicular (QRS interval) conduction times are prolonged (P less than 0.001), as well as the RRF of His-Purkinje system. The maxium increase of AH,HV and QRS intervals appears 3 minutes after drug administration; at the 5th minute these intervals have already slightly decreased, then they decrease almost quickly till the 15th minute and more slowly till the 30th minute. The absolute prolongation of the HV and QRS intervals is more marked in the patients in which these intervals are more prolonged during the control study, while the percentage increase is almost equal in the patients with normal HV and QRS interval and in those with prolonged HV and QRS intervals. The electrophysiological properties of the drug are discussed on the basis of the obtained results.


[Hypoplasia of left pulmonary artery secondary to aorta-right pulmonary artery anastomosis. Diagnostic and therapeutical considerations (author's transl)]. Three cases of tetralogy of Fallot treated under twelve months of life with aorta-right pulmonary artery anastomosis are reported, in which severe hypoplasia of left pulmonary artery secondary to palliation was present. In the first two cases this complication was not recognized during investigation prior to repair and both died of low cardiac output due to severe right ventricular hypertension. In the third case the complication was evidenced and treated with interposition of a prosthetic valved conduit between right ventricle and confluence of the pulmonary arteries. Hypoplasia or atresia of left pulmonary artery and right pulmonary vascular obstructive disease, secondary to a long-functioning aortopulmonary anastomosis, require surgical consideration identical to tetralogy of Fallot with single pulmonary artery, which is corrected with the use of a valved conduit. The Authors suggest that aortopulmonary shunts should be dismantled within two years and investigation prior to repair should include: 1) exploring of all cardiac chambers and right pulmonary artery through the fistula; 2) injection of contrast medium in right pulmonary artery, aortic root, right ventricle or pulmonary trunk. These diagnostic methods should demonstrate continuity between right ventricle and pulmonary arteries, confluence of pulmonary arteries, and rule out acquired left pulmonary artery atresia, right pulmonary vascular obstructive disease, right ventricular outflow tract atresia.


[The echographic aspects of the foetal heart (author's transl)]. In 15 pregnant women, during the last month of pregnancy, the heart of living human foetuses has been searched for with echocardiological techniques, using a multiple-crystal transducer. The single heart structures distinguished on the bidimensional image were picked out with a single ultrasonic ray and photographed with a Polaroid camera. The echocardiological image revealed of such structures is similar to that found in newborns. The aorta and the mitral valve have been registered in all cases. It has been possible, with minor frequency, to reveal the echoes of the tricuspid valve, of the interventricular septum, of the posterior wall of the left ventricle and of the pulmonary valve. The left atrium/aorta root relationship resulted almost always higher with regard to the values considered normal for newborns. The behaviour of the interventricular septum is of particular interest, as it always showed a normal movement. This report indicates that the functional load of the different heart cavities during the various phases of interuterine life is in perfect equilibrium, notwithstanding the substantial anatomical and functional modifications which occur during the course of foetal life. The Authors maintain that echocardiological methods could become indispensable in understanding the physiological and pathological aspects of foetal circulation.


[Echo-phonomechanocardiographic relations of the systolic time intervals of the left ventricle (author's transl)]. 15 normal subjects and 23 subjects with cardiac valve disease have been studied. All the subjects have undergone an echo-phonomechanocardiographic investigation with simultaneous recording of the tracings. The values of the systolic time intervals have been obtained from the echo-cardiographic tracings and compared with those obtained from the phonomechanocardiographic tracings evaluating the differences between them. The comparison has pointed out values nearly coincident, providing useful elements about the relations existing between the cardiac sounds and the moments of closure of the mitral and aortic valves. Moreover, the two techniques are considered useful and valid in the evaluation of the left ventricle performance by the systolic time intervals, especially when the recording is carried out simultaneously.


The usefulness of the phonocardiography in the diagnosis of the extrinsic obstruction of the pulmonary artery. Report of a case (author's transl). The Authors describe a case in which an anterior mediastinal tumor manifested as a cardiovascular disease. Phonocardiographic findings and their modifications induced by respiratory manoeuvres are presented and discussed. Particular enphasis is given to the disappearance, with deep inspiration, of the pulmonary ejection murmur associated with the simultaneous paradoxical behaviour of pulmonary component of the second sound. The Authors suggest that such findings can be indicative of pulmonary outflow obstruction by an external mass.


[Variations of the pre-coronary times (1973-1977) (author's transl)]. A comparison has been made between the pre-coronary times of IMA patients in the UCIC of the S. Camillo Hospital in Rome between 1973 and 1977, and the pre-coronary times of patients in another UCIC in the same city, in order to record possible modifications. From 1973 to 1977 there has been a decrease in the pre-coronary times: this is the result of a better hospitalization policy and more frequent admission directly to the UCIC. The mortality rate according to the data in question, has also decreased from 21% in 1973 to 10% in 1977, with a marked reduction for patients with a pre-coronary time superior to 12 hours. This is also connected to the fact that there is a considerable reduction in the number of patients, generally suffering from serious complications, transferred to UCIC from other wards. From an analysis of the data we infer that there has been an improvement in the policy governing the admission of patients to UCIC, a policy which could be improved further by a more adequate training both for doctors and the population in general.


Differentiation between patients with malignant diseases and non-malignant diseases or healthy donors by changes of fluorescence polarization in the cytoplams of circulating lymphocytes. Changes in the degree of fluorescence polarization in the fluorescent cytoplasm of lymphocytes from patients with malignant diseases, patients with nonmalignant diseases, and normal donors were investigated by the fluorescence polarization technique. In order to confirm Cercek's cancer test, two kinds of parameters, the degree of fluorescence polarization of lymphocytes stimulated with phytohemagglutinin (PHA) and cancer basic protein, were obtained for each person. Patients with malignant diseases were easily differentiated from the others without exception by an index, the response ratio of the degree of fluorescence polarization. Usefulness of this technique was confirmed for the detection of cancer-bearing patients.


Effect of Ehrlich ascites tumour on the clinical course and pathology of mice having carbon tetrachloride-induced hepato-renal necrosis. Ehrlich ascites tumour, administered intraperitoneally 18 hr following a dose of CCl4 by stomach tube, produces and irreversible illness characterised by failure to resolve extensive hepato-renal necrosis. Equally extensive hepato-renal necrosis occurs in animals given CCl4 and saline; such animals, however, remain clinically well and show rapid histological regeneration of both organs. Carbon tetrachloride given to mice on day 5 of tumour growth produces hepatic necrosis only, the kidney of such animals being immune to the necrotising effect of CCl4. Such animals remain well, and histological recovery of the liver is rapid. It is proposed that the Ehrlich tumour produces a "regeneration-inhibiting" toxin, active against the damaged liver and kidney; the CCl4-damaged kidney fails to excrete this toxin, hence the irreversibility of hepato-renal damage and a fatal outcome. A marked reduction of ascites volume as compared with controls was observed in those animals given Ehrlich tumour 18 hr after CCl4, but not in animals given CCl4 on day 5 of Ehrlich ascites tumour growth.


Late effects of fetal mice x-irradiated at middle or late intrauterine stage. Mice of F1 hybrids of (C57BL/6xWHT) strains were exposed in utero to 200 R of X-rays at 12 or 16 approximately 18 days post coitum. Mice of both sexes were allowed to liver through their life span, and the induction of tumors, growth retardation, and induction of degenerative diseases were examined. A significant enhancement of lung, pituitary gland, and ovary tumorigenesis was observed in mice irradiated at the late intrauterine stage. Also, incidence of liver and skin tumors was increased slightly in this group, whereas thymic lymphomas were not induced. Persistent growth retardation, several congenital malformation, and amyloid degeneration were found in mice irradiated at the middle intrauterine stage. However, no increases incidence of tumors was seen in this group. Moreover, incidnece of lymphoreticular tissue tumors, lung tumors, and leiomyosarcomas of uterus was significantly decreased by irradiation at  the middle intrauterine stage from the unirradiated control and the late intrauterine irradiated group.


Correlation of differences in antigenicity of four 3-methyl-cholanthrene-induced tumors in syngeneic mice with the susceptibility of tumors to an immunopotentiator, PS-K. Eleven syngeneic tumors were induced by 3-methylcholanthrene in DDD mice. Their antigenicity in vivo was examined by measuring growth inhibition of transplanted tumors in mice immunized by implanting tumors into their tail and then cutting off the tail. Four tumors with different antigenicity were selected from them for study. The growth-suppressive effect of an immunotherapeutic agent, PS-K, on the four tumors was examined. The growth of highly antigenic tumors in mice was inhibited by PS-K, but that of weakly antigenic tumors was not, and a close correlation was found between the antigenicity of tumors and the antitumor activity of PS-K against them. No similar correlation was found between their antigenicity and their sensitivity to a mitotic poison, 6-mercaptopurine. These results suggest that the antitumor effect of an immunopotentiator depends greatly on the antigenicity of tumors.


Effect of combined use of anticancer drugs with a polysaccharide preparation, Krestin, on mouse leukemia P388. When CDF1 mice inoculated intraperitoneally with leukemia P388 were treated intraperitoneally with mitomycin-C, of which dose was not so effective, on day 1 and given intraperitoneally polysaccharide preparation, Krestin, on day 2, 5, or 8 after tumor inoculation, best therapeutic results judged from 60-day survivors were obtained by combined use of mitomycin-C on day 1 and Krestin on day 5. Besides mitomycin-C, none of anticancer drugs such as cyclophosphamide, 5-fluorouracil, and 6-mercaptopurine was effective in this system. From these results, it was concluded that in this system, enhancement of anti-tumor effect of mitomycin-C by Krestin depends on the time of Krestin administration and that the combination effect is observed with mitomycin-C but not with cyclophosphamide, 5-fluorouracil, or 6-mercaptopurine.


Cytotoxic activity of human blood monocytes against cultured breast cancer cells. Cytotoxic activity of blood monocytes from 19 healthy individuals was examined in vitro against the cultured floating cells of human breast cancer origin. Target tumor cells inoculated on monocyte cultures and maintained for 5 days were reduced in number, compared with the control cultues of tumor cells alone. Increased cytotoxicity of the monocytes was noted after in vitro activation by heat-killed Corynebacterium liquefaciens (Propionibacterium acnes). C. liquefaciens of the same dose as used for monocyte activation had no effect on the tumor cell growth. These results may indicate that blood monocytes have an antitumor activity and it can be strengthened by treatment with C. liquefaciens. Antitumor activity of blood monocytes and macrophages was discussed on a concept of the mononuclear phagocyte system. A possible development of a new assay method for macrophage function as a parameter of host defence state against cancer will be expected.


Antitumor activity of water-soluble platinum(II) complexes of 1,2-cyclohexanediamine isomers. Dichloroplatinum(II) complexes of 1,2-cyclohexanediamine isomers were reported to have high antitumor activity against leukemia L-1210 and P-388, but they were hardly soluble in water and, therefore, dibromo- and diiodo-platinum(II) complexes of 1,2-cyclohexanediamine isomers were prepared. They were not soluble in water but showed high antitumor activity. When the chloride ions are replaced with sulfate ions, they become soluble and also they are anti-tumor active. Bismonobromoacetatoplatinum (II) complexes were synthesized and tested. Mono- and bis-(D-glucuronato)platinum(II) complexes of 1,2-cyclohexanediamine isomers were newly synthesized. All of them were freely soluble in water and showed high antitumor activity with low toxicity. The chelated mono(D-glucuronato)platinum(II) was found to be more effective than the monodentate bis(D-glucuronato)platinum (II) complexes.


Evidence for a defective inhibition of pentagastrin-stimulated gastric acid secretion by antral distension in the duodenal ulcer patient. Graded antrum distension by 50-, 100-, and 150-ml balloon volumes significantly stimulated gastric acid secretion in 6 duodenal ulcer patients, but had no stimulatory effect in 6 healthy subjects. Antrum distension by 150-ml balloon volume had no effect on the acid response to submaximal stimulation by a continuous intravenous infusion of pentagastrin in the duodenal ulcer patients, but significantly inhibited this response in the healthy subjects. The inhibition by antrum distension in the healthy subjects amounted to 20%. The results suggest that antrum distension elicits stimulatory as well as inhibitory mechanisms in man with inhibition of acid secretion predominating in healthy subjects and stimulation of acid secretion predominating in duodenal ulcer patients. The mechanism of inhibition is unknown.


Cell wall-defective variants of pseudomonas-like (group Va) bacteria in Crohn's disease. Several animal transmission studies have indicated that Crohn's disease may be caused by a filterable agent. Filtrates of homogenized tissues were prepared from 8 patients with Crohn's disease, 9 patients with chronic ulcerative colitis, and 20 control patients without inflammatory bowel disease. Conventional bacteriological cultures and hypertonic cultures for cell wall-defective bacterial variants were performed on the filtrates. Bacterial revertants (parent forms) of cell wall-defective variants were obtained from filtrates of various tissues including mesenteric lymph nodes of all patients with Crohn's disease. In no instance were revertants cultured from tissue filtrates of the other patients. The 11 revertants isolated from the 8 patients were identified as Pseudomonas-like bacteria, most closely identifiable with group Va.


Effects of caffeine and coffee on fluid transport in the small intestine. Although caffeine is known to stimulate small intestinal water secretion in man, no available studies have examined the effect of coffee on small intestinal fluid transport. We have studied the effect of both coffee and caffeine on jejunal sodium and water transport by the triple lumen intestinal perfusion technique. Both caffeine and coffee solutions had equivalent caffeine concentrations, approximating the caffeine concentration in one cup of coffee. Control and coffee perfusion resulted in net absorption of 0.9 +/- 0.2 and 1.1 +/- 0.3 ml per min per 30 cm, respectively, whereas caffeine perfusion resulted in net water secretion of 2.4 +/- 0.4 ml per min per 30 cm (P less than 0.02). Net sodium absorption was observed with control and coffee solutions with a mean absorption of 190 +/- 32 and 184 +/- 40 muEq per min per 30 cm, respectively. Net sodium secretion of 334 +/- 25 muEq er min per 30 cm was observed during caffeine perfusion (P less than 0.02). We conclude that coffee perfusion has no significant effect on sodium and water transport in the jejunum in spite of the marked secretory effects of caffeine solutions. The results of this study yield further support to the suggestion that the effects of caffeine may not be similar to the effects of caffeine-containing compounds such as coffee.


Multiple immunoreactive forms of vasoactive intestinal peptide in human colonic mucosa. Extracts of human colon mucosa and muscle were fractionated by gel filtration and ion exchange chromatography and molecular forms of vasoactive intestinal peptide (VIP) estimated by radioimmunoassay. Total concentrations of immunoreactive VIP in mucosa were 237.1 +/- 53.9 and 119 +/- 26.0 pmoles per g in muscle. In muscle, over 90% of the immunoreactivity was accounted for by a single component indistinguishable from porcine VIP by gel filtration and ion exchange chromatography. In contrast, mucosa contained four components with VIP-like immunoreactivity. One of these had the chromatographic properties of porcine VIP; the others were less positively charged. Two of the new VIP-like components had gel filtration properties similar to those of VIP and are, therefore, probably of similar size to the porcine octacosapeptide, and the remaining component emerged later on Sephadex and is, therefore, probably a smaller peptide. Immunoreactive VIP in muscle extracts is believed to originate from nerve fibers, and this form of VIP is likely to be identical to the previously characterized form of VIP. However, immunoreactive VIP in mucosal extracts may originate either from endocrine cells or nerve fibers. The possibility arises, then, that there are different immunoreactive forms of VIP in nerves and endocrine cells.


Evaluation of xylose absorption as measured in blood and urine: a one-hour blood xylose screening test in malabsorption. The assessment of D-xylose absorption by measurement of urinary excretion is subject to several sources of error. An investigation has been made into the use of the blood levels obtained after a 5-g dose as an assessment of malabsorption. We have shown that optimal discrimination between normal and impaired xylose absorption is achieved by measuring blood xylose levels 1 hr after oral administration and then correcting the results to a constant body surface area. The reference range was 0.65 to 1.33 mmoles per liter for values corrected to 1.73m2. In a prospective study of the value of this modification in investigating malabsorption, the incidence of false-negatives and false-positives was found to be 4.8 and 2.2%, respectively. The test is of particular value in the elderly because surface area corrected 1-hr blood xylose levels are independent of age, eliminating the expected and observed declines in renal function and urinary xylose excretion in this age group.


Effects of pentagastrin, G17, and G34 on the electrical and mechanical activities of canine antral smooth muscle. Intracellular microelectrode technique and standard organ bath technique were used to investigate the effects of pentagastrin, G17, and G34 on the electrical and mechanical activities of canine antral circular muscle. All three molecular forms increased the amplitude and duration of the plateau of the gastric action potential and the frequency of spontaneous action potentials. They also increased the amplitude and frequency of spontaneous contractions. G17 was equal to or less potent than pentagastrin in all of its actions on this tissue. G34 had an equal or greater activity than G17. The electrical studies indicate that G17 is active in this tissue in a physiological range of concentrations. The ED50 for the effect of G17 to increase the amplitude of the plateau potential is less than that for the effect of G17 on gastric secretion, indicating that this is a physiological action of gastrin. Atropine studies indicate that only part of the in vitro inotropic action of gastrin is caused by the release of acetylcholine from nerve terminals, but that the chronotropic action is attributable to a direct effect on the smooth muscle membrane.


Influence of high luminal PO2 on the anaerobic bacteria of the rat colon. The influence of high luminal PO2 on colonic anaerobes was investigated by chronically perfusing the colon of unrestrained rats with O2 or N2. A luminal PO2 of approximately 700 mm Hg did not significantly influence the fecal concentration of total anaerobes recovered on rabbit blood agar or of Bifidobacterium sp. and Bacteroides sp. recovered on selective media. However, the O2 infusion reduced the rat's excretion of CH4 (a metabolic product of anaerobic bacteria) to about 5% of the control value, although the feces of O2-treated rats produced CH4 normally. This study suggests that the production of metabolites by the colonic bacteria may sometimes be more accurately studied in situ rather than in culture. Measurement of volatile bacterial metabolites in breath may provide a simple means of studying the in situ metabolism of a variety of colonic bacteria in man.


Histofluorescence of 3-methylcholanthrene metabolites in the rat pancreas. Pancreases of Long-Evans rats were treated in vitro and in vivo with 3-methylcholanthrene and examined by histofluorescence techniques. For in vitro studies, tissue sections were dipped in 3-methylcholanthrene and incubated in vitro. These sections revealed that acinar and ductal epithelium had equal fluorescence which suggested equal metabolic capability. For in vivo experiments, 3-methylcholanthrene was injected intraperitoneally and the rats were killed 60 min later. Two additional rats were provided with biliary cannulae to divert bile from the pancreatobiliary ducts. Frozen sections, 16 mu in thickness, were prepared and examined under the fluorescence microscope. The sections revealed that fluorescence was concentrated in the epithelium and lumen of ducts. Animals with diversion of bile from the pancreatobiliary ducts had similar intensity and distribution of fluorescence. The in vivo studies showed that ductal epithelium was exposed to greater concentrations of carcinogen than nonductal epithelium. These observations provide a link between epidemiological studies that show an increased incidence of pancreatic adenocarcinoma in populations exposed to environmental carcinogens and morphological studies that show the ductal cell to be the most likely cell of origin.


Phalloidin-induced cholestasis in the rat: relation to changes in microfilaments. The influence of phalloidin, an agent that causes irreversible polymerisation of actin into microfilaments, on bile secretion and hepatocyte ultrastructure was examined in rats. Phalloidin was given intraperitoneally at the dose of 50 microgram per 100 g of body weight per day for 1, 3, or 7 days. The following was observed. (1) Bile flow decreased, as compared to controls, by 19% after 1 day, 34% after 3 days, and 55% after 7 days. Bile acid secretion was also decreased. (2) Electron microscopic examination of the hepatocyte in treated animals revealed an increased thickness of the pericanalicular microfilamentous network and a dilatation of bile canaliculi. Stereological examination revealed an increase in the relative volume of the microfilamentous network (per unit of hepatocyte cytoplasm) of 2.55% after 1 day, 4.06% ater 3 days, and 6.16% after 7 days. (3) [14C]Erythritol biliary clearance, measured after 7 days, decreased in parallel to bile flow, suggesting that the decrease in bile flow was of canalicular origin. [14C]Sucrose biliary clearance increased in treated animals, suggesting an increased permeability of the biliary system to sucrose. There was a predominant decrease in the bile acid independent bile flow. These data provide circumstantial evidence for the hypothesis that microfilament dysfunction can produce cholestasis.


Cell-mediated cytotoxicity of desialylated human lymphocytes induced by a mitogenic mammalian liver protein. The ability of a purified rabbit liver membrane protein that selectively binds desialylated glycoproteins to induce desialylated human peripheral blood lymphocytes to mediate mitogen-induced cellular cytotoxicity has been determined. After short term exposure to purified hepatic binding protein, desialylated, but not intact, lymphocytes exhibited mitogen-induced cellular cytotoxicity against Chang target cells, which do not have exposed hepatic binding protein on their surface membrane. Furthermore, desialylated lymphocytes, in the absence of purified hepatic binding protein, reduced the proportion of isolated rabbit hepatocytes that adhered to plastic to a significantly greater extent than did intact lymphocytes, suggesting that the exposed hepatic binding protein demonstrated on the surface membrane of these target cells is capable of inducing mitogen-induced cellular cytotoxicity. The specific inhibition of this effect by asialo-orosomucoid, but not by intact orosomucoid, indicates that the site involved in the binding of asialo-glycoproteins to hepatic binding protein is probably also responsible for the induction of mitogen-induced cellular cytoxocity. The demonstration that hepatic binding protein, a normal constituent of the surface, membrane of mammalian hepatocytes, can induce mitogen-induced cellular cytotoxicity, suggests that mitogen-induced cellular cytotoxicity may be a mechanism of cellular injury in vivo.


Hepatic cholesterol ester hydrolase in human liver disease. Human liver contains an acid cholesterol ester hydrolase (CEH) of presumed lysosomal origin, but its significance is unknown. We developed a modified CEH radioassay suitable for needle biopsy specimens and measured hepatic activity of this enzyme in 69 patients undergoing percutaneous liver biopsy. Histologically normal livers hydrolyzed 5.80 +/- 0.78 SEM mumoles of cholesterol ester per hr per g of liver protein (n, 10). Values were similar in alcoholic liver disease (n, 17), obstructive jaundice (n, 9), and miscellaneous hepatic disorders (n, 21). In contrast, mean hepatic CEH activity was more than 3-fold elevated in 12 patients with acute hepatitis, 21.05 +/- 2.45 SEM mumoles per hr per g of protein (P less than 0.01). In 2 patients studied serially, CEH returned to normal as hepatitis resolved. CEH activity in all patients paralleled SGOT levels (r, 0.84; P less than 0.01). There was no correlation with serum levels of free or esterified cholesterol nor with serum activity of lecithin-cholesterol acyltransferase, the enzyme responsible for cholesterol esterification in plasma. These studies confirm the presence of CEH activity in human liver and show markedly increased activity in acute hepatitis. The pathogenesis and clinical significance of altered hepatic CEH activity in liver disease require further study.


Incidence and clinical significance of sinusoidal dilatation in liver biopsies. In a review of 906 consecutive liver biopsies, sinusoidal dilatation, unrelated to passive congestion of the liver, sinusoidal infiltration, or cirrhosis, was found in 26 cases (2.9%). In 21 of them the final diagnosis was a neoplastic or granulomatous disease (tuberculosis, brucellosis, Crohn's disease), but in only half of them was there evidence of neoplastic or granulomatous infiltration of the liver. In the remaining cases, sinusoidal dilatation was either the only histological abnormality or it was associated with nonspecific changes. Although the pathogenesis of sinusoidal ectasia is not known, our findings indicate a definite relationship to the presence of tumor or granulomatous disease in the liver or elsewhere in the body. It is concluded that the finding of sinusoidal dilatation as an isolated histological change in a liver biopsy specimen should prompt the search for a tumor or a disease associated with granulomas.


Effect of age and parenchymal liver disease on the disposition and elimination of chlordiazepoxide (librium). There is an increased incidence of unwanted sedation associated with chlordiazepoxide usage in the elderly and in patients with liver disease. To determine whether pharmacokinetic alterations could account in part for these observations we studied the disposition and elimination of intravenously administered chlordiazepoxide in 27 healthy controls aged 16 to 86 years, 8 patients with cirrhosis, and 5 patients with acute viral hepatitis. Both increasing age and parenchymal liver disease led to similar changes in chlordiazepoxide pharmacokinetics. Over the age range 20 to 80 years, elimination half-life (t1/2(beta)) increased from 7 to 40 hr (r, 0.67; P less than 0.001) attributable to a decrease in plasma clearance from 30 ml per min to 10 ml per min (r, -0.71; P less than 0.001) and an increase in volume of distribution from 0.26 to 0.38 liters per kg (r, 0.60; P less than 0.05). Similarly, a decrease in plasma clearance in cirrhosis (7.7 +/- 2.1 compared to 15.3 +/- 4.4 ml per min, P less than 0.01) and acute viral hepatitis (6.1 +/- 4.3 compared to 18.1 +/- 7.1 ml per min, P less than 0.01) relative to age-matched controls and an increase in the volume of distribution resulted in a prolongation of the elimination half-life in both forms of liver disease. Impaired elimination of chlordiazepoxide may account in part for the increased incidence of oversedation seen in the elderly and in patients with liver disease.


Iproclozide fulminant hepatitis. Possible role of enzyme induction. The authors report the cases of 3 patients who died from fulminant hepatitis after receiving iproclozide, a hydrazine-containing monoamine oxidase inhibitor. Fulminant hepatitis in these patients resembled that reported in patients receiving other hydrazine-containing monoamine oxidase inhibitors: (1) the 3 patients were women; (2) the monoamine oxidase inhibitor has been ingested for 1 month or more; (3) the main clinical manifestations were jaundice and disorders of consciousness; (4) hypersensitivity manifestations were absent; (5) the predominant liver lesion was necrosis; (6) all 3 patients died. In our 3 patients, jaundice occurred 7 to 10 days after the adjunction to iproclozide of a microsomal enzyme inducer. These observations suggest that concomitant administration of iproclozide and of microsomal enzyme inducers may produce fulminant hepatitis in man. It is speculated that iproclozide could be, like iproniazid, transformed into a hepatotoxic metabolite, the production of which would be increased by microsomal enzyme induction.


Gastric and esophageal dysfunction after ingestion of acid. Esophageal and gastric function was measured in a patient who swallowed a household acid solution. Dysphagia, transient ulceration of the esophagus with luminal narrowing, and complete loss of peristalsis without loss of lower esophageal sphincter function were noted. Gastric dysfunction appeared 2 weeks after ingestion with complete obstruction, necessitating antral resection. The proximal stomach was relatively spared.