The activity of alkaline phosphatase in the process of regeneration of rat liver. Using GOMORI'S technique, the present authors investigated the dynamics of the alkaline phosphatase activity in the process of liver regeneration after partial hepatectomy. In all 80 rats of the Wistar strain were subjected to experiment, 60 to 75% of the liver parenchyma being removed from each of them. In the course of regeneration a gradual increase in the enzyme activity was observed within the first 48 hours following the operation. This was succeeded by a slow decline of the activity, and after the 25th day after the operation the reaction intensity resembled that recorded for the control animals. It was also ascertained that the fatty degeneration of the liver noted in the initial period of regeneration does not inhibit the activity of alkaline phosphatase.


The electron-microscopic picture of postnatal development of oligodendroglia. Ultrastructural studies were carried out on the oligodendroglia in the corpus callosum of rat brain in different periods of the animal's extrauterine development. These investigations showed a possibility of distinguishing in the early stages of ontogenesis three forms of oligodendroblasts differing from one another by electron density and the organelle content of the cytoplasm. The beginning of the production of medullary sheaths was noted as early as the first day after birth and the termination of the morphological phase of this process was recorded on the 8th day of postnatal ontogenesis. Moreover, it was found that on the 17th day of the rat's life the process of differentiation of oligodendroglia in the corpus callosum comes to an end.


Treatment of the transsexual and medicolegal issues. The carnival, stage or masquerade ball are the only situations where a male is tolerated in assuming the femal role, or vice-versa in Western Society. The transsexual, finding it impossible to maintain his or her self-esteem, is given the opportunity to resolve a profound dilemma via this new medical procedure of six reassignment. Endocrine therapy and conversion surgery raise medicolegal issues illustrative of the difference in perspective between medicine and the law. The nonjudgmental tradition of medicine and the judgmental and prosecuting tradition of law produce attitudes difficult to reconcile in dealing with legal issues that arise in transsexual treatment. This paper deals with the legal ramifications of cross-dressing as part of treatment, transsexual surgery and its exposure to the possibility of legal prosecution, and finally documentary alterations such as birth certificate changes.


Deaths resulting from drugs of abuse.

Deaths resulting from drugs of abuse. This report covers 40 narcotics fatalities from the Northbaden are (Federal Republic of Germany) that came to the Institute of Forensic Medicine, Heidelberg University for autopsy in the years of 1968-1973. Information is given on pathological anatomic and toxicologic findings. A biographical, sociological, psychiatric and criminological catamnesis gave important insight in determinants, motivations, development and course of drug dependence in this area. Basic details are shown and critically evaluated.


Polymorphism of A, B and H substances in seminal fluid. The polymorphism of A, B and H substances recently described in human saliva has been studied in seminal fluid of 71 subjects. Semen was found to contain the same molecular species of A, B and H substances which were observed in saliva and red cells. The comparison of gel-chromatographic patterns of the saliva and semen of 40 subjects showed identical ABH patterns in 26 cases, while in the remaining 14 some differences were observed.


On the significance of posterior crico-arytenoid muscle hemorrhage. Sixteen cases of gross fresh PCA muscle hemorrhages were collected from a series of 1500 medicolegal autopsies consecutively performed by the authors, an incidence of about 1%. These hemorrhages occurred in cases of deaths resulting from a variety of causes, including natural disease, physical agents and drugs. In none of the cases was there any evidence of external trauma to the neck. In two cases endotracheal intubation had been performed; however, the lack of injury to the tissues between the tube and the muscles probably rules this factor out. In over 50% of the cases, PCA muscle hemorrhages were found in association with "asphyxial" petechial hemorrhages. A consideration of our findings and a study of the literature points out that before considering any individual case of PCA muscle hemorrhages as resulting from direct trauma, additional evidence is needed.


[Allergen-specific immunoglobulin E in the serum (RAST), skin tests and mucosa provocation tests using house dust and house mite extract in rhinopathy and bronchial asthma]. Allergy against house dust and house dust mite was examined by RAST, skin and provocation test, with not identical allergen extracts. 50 patients with allergic rhinitis showed insufficient RAST results with house dust, better results for house dust mite. 65 patients with allergic asthma bronchiale presented satisfying results with house dust, good results with house dust mite. Recommendations are given for the application of RAST in the diagnosis of allergy.


Forensic odontology in identification of a murder victim in India. Absence of 2 left upper incisors and their sockets in the skull of a victim helped to pinpoint identity in a murder case. Age at death of the victim was calculated using Gustafson's method.


Self-inflicted bite marks in battered child syndrome. Examination of a child abuse victim revealed bite marks on both arms. These lesions are usually pursued as a clue to the identity of the specific perpetrator. We demonstrated that the bite was from the victim. Importance of this phenomenon in evaluation of bite injuries is discussed.


[Homotransplantation of the spleen].

[Homotransplantation of the spleen]. A report of experiences and results on 63 homotransplantations of spleen in dogs is given; 50 of the transplantations were performed in an orthotopic and 13 in a heterotopic way. 10 of the latter had undergone an earlier splenectomy and 3 an earlier orthotopic transplantation, followed by a complete resorption of the first transplant. The course of the transplant was observed by scintigraphy, the function of the anastomosis was controlled by angiography. The immunological and hematological changes were analyzed together with the changes of the protein-electrophoresis and those of the fibrinogenic factors. Scintigraphy has proved to be very useful for the diagnosis of transplant rejection; the measurement of the activity of the lysozyme in the serum compared with the pathoanatomical findings showed a characteristic increase of the activity of the lysozyme during the rejection crisis.


[Immunology of malignant melanomas].

[Immunology of malignant melanomas]. The intraperitoneal diffusion-chamber-technique is a closed cell culture system in vivo. Hamster melanoma cells were capable in iso- and heterologous host animals for exponential proliferation. When the host animals had been preimmunized with melanoma cells or hamster lymphocytes a rapid cell lysis was observed in the chambers within 4 hrs after implantation. The cell numbers were reduced about 1-10% of the inoculum. After 6-10 days the cell numbers in chambers from immunized hosts slowly increased in spite of a high level of cytotoxicity in the serum of the animals.


Living will does not resolve medical-ethical-legal dilemma. The Living Will is an ethically acceptable document that serves to convey a patient's attitude toward terminal illness and to facilitate a physician's decision not to prolong the dying process. But in a Christian context, the Christian Affirmation of Life is a superior document by which to express and aid such decision making. Because the Living Will advances the concept of negative euthanasia--an ethical, legal, and political misnomer--and abets the effort to legalize positive or direct euthanasia, it should not be given legal recognition. As a legal document, the Living Will could seriously affect both patients and physicians.


Ultrastructure of osteoid osteoma.

Ultrastructure of osteoid osteoma. The ultrastructure in five cases of osteoid osteoma is described. The osteoblasts generally had a morphology similar to that of normal osteoblasts with a few differences. They contained irregular indented nuclei, glycogen particles, abundant fine intracytoplasmic fibrils, and rare iron containing lysosomes. In several osteoblasts in two cases there were atypical mitochondria with a lobulated or "honeycomb" appearance. These atypical mitochondria were also observed in two osteoclasts; otherwise the cells resembled normal osteoclasts. Other cells present in osteoid osteoma besides osteocytes included osteoprogenitor cells resembling Scott type A and B cells and cells in transitional stages of differentiation. The osteoblasts most likely originated from Scott type A cells of preosteoblasts. The areas of mineralized matrix conformed to the morphology of coarse woven bone. Varying amounts of osteoid were noted. In two cases the osteoid contained, in addition to collagen, fine granular material, which probably represents polysaccharides. No nerve fibers were identified in the material studied. It is thought that osteoid osteoma is a benign neoplastic lesion. A case of osteoblastoma was studied for comparison; the osteoblastoma cells were found to have basically the same morphology as the cells in osteoid osteoma, including the atypical mitochondria. Our ultrastructural observations support the idea that osteoid osteoma and osteoblastoma are closely related  lesions.


Renomedullary interstitial cell lesions and hypertension. The notion that renomedullary nodules of the kidney arise as a response to hypertension was tested. Heart weight, ratio of heart weight to body weight, and blood pressure in 160 patients in whom such nodules were found at autopsy were compared with similar measurements in 160 patients without  nodules. There were no significant differences between the two groups, and it was concluded that there was no evidence to support the idea that the renomedullary nodule represents a response to hypertension.


Malignant hypertension in blacks. Malignant intrarenal arterial disease as observed by light and electron microscopy. Volhard and Fahr recognized that hypertensive intrarenal vascular disease could be divided into two groups corresponding to the clinical states of benign and malignant hypertension. Since that time, numerous papers on malignant hypertension, primarily dealing with European derived populations, have emphasized fibrinoid necrosis of small arteries and arterioles as the lesion of malignant hypertension, although some have also recognized a myxoid intimal lesion as characteristic. Today in the United States, however, a significant proportion of malignant hypertension occurs in blacks. In the present study, patients have lacked fibrinoid necrosis of arterioles and only rarely have had some atypical necrosis of small arteries. The prominent, but not pathognomonic, lesion in this series is a myxoid intimal thickening of small arteries consisting of smooth muscle cells, acid mucopolysaccharides, basement membrane-like materal, collagen, and other amorphous and unidentified material, probably plasma derived.


Frequency of chromosomal fluorescence polymorphism in normal persons and in clinical patients with diagnosed chromosome aberrations. By means of a computer program the frequencies of the strongly fluorescent polymorphous chromosomal segments on chromosomes Nos. 3, 4, 13, 14, 15, 21, and 22 among 89 random normal persons and 247 persons suspected of having various chromosome aberrations were determined. It was discovered that: 1. In none of the 13 diagnosis categories are divergencies in frequency of autosomal fluorescence polymorphism, as compared to the normal group, statistically determinable. 2. A worthwhile comparison of the various frequencies of fluorescence polymorphism as recorded by the various investigators in not possible at present, since the applied methods of assessment differ too widely. 3. Standardization of the criteria of assessment and of the nomenclature for the polymorphous chromosomal segment would seem to be a matter of urgent necessity.


Ss blood group associated PAS-staining polymorphism of glycoprotein 3 from human erythrocyte membranes. It is shown by discontinuous sodium dodecylsulfate (SDS) polyacrylamide gel electrophoresis of human red cell membranes, followed by periodic acid Schiff (PAS) staining and densitometry, that the band PAS-3 (monomeric Ss glycoprotein) exhibits a polymorphism with respect to its staining intensity. In membranes of the genotype SS the staining intensity of this band is about 1.5 times higher than in ss membranes. The experimental error of the method does not permit one to decide, whether membranes of blood type Ss exhibit an intermediate staining intensity or not. The SDS electrophoretic molecular weight of PAS-3 is the same for SS, Ss, or ss membranes. The ratio of PAS to coomassie blue staining intensities, which are a measure of sialic acid and protein content, respectively, are equal for PAS-3 in glycoprotein preparations from SS and ss erythrocytes. These data indicate that the above phenomenon reflects a difference in the glycoprotein content between SS and ss membranes.


Partial and complete trisomy 9: delineation of a trisomy 9 syndrome. Two infants with trisomy involving chromosome 9 are described. One had complete trisomy 9 and the other karyotype 47,XX,+der(9),t(7;9) (p22;q32)mat. A trisomy 9 syndrome is delineated, consisting of features of the trisomy 9p syndrome and various other malformations. These include abnormalities of the cardiovascular and urogenital systems, cranial suture anomalies, dislocation of the hips and knees and early death. A possible relationship of some of these findings to regions of 9q involved in cases of partial trisomy 9 is suggested.


Chromosome measurements on an XXp+ male. Measurements carried out on an XX male show that there is a significant difference between the lengths of the short arm of the 2 X chromosomes of this patient and that the short arm of the longer X is also significantly different from that of the X chromosomes derived from normal individuals. The difference lies specifically in the size of the terminal band of the short arm. These results are consistent with the X-Y interchange hypothesis of Ferguson-Smith.


Distribution of spontaneous chromosome breaks in human chromosomes. Localization of chromosome breaks in human chromosomes was analyzed in 264 peripheral lymphocyte cultures. Three hundred and sixty-nine chromosome breaks could be exactly localized to a chromosome band or region of the Paris Conference nomenclature. The distribution of breaks in the chromosome regions was found to be nonrandom. Chromosome 3 alone had 23% of the breaks and region 3p2 had 13% of the total breaks. Some other chromosome regions, such as 5p1, 9q1, 14q2, and 16q2 also displayed clustering of breaks. Sex chromosomes had less breaks than expected. Spontaneous chromosome breaks were almost exclusively located in the lightly stained G bands.


Morphologic variability of human chromosomes: polymorphism of constitutive heterochromatin. Polymorphism of constitutive heterochromatin has been studied in a series of 30 normal individuals. A high frequency of C-band variants were observed. Twenty-six of the 30 individuals studied had at least one polymorphic variant of the C band. A total of 42 variants were recorded which were predominately localized near the centromeric heterochromatin block of chromosome 9 (26.19%), chromosome 16 (19.05%), and chromosome 1 (16.66%). These results are discussed together with the findings revealed by different studies.


GD (--) Aachen, a new variant of deficient glucose-6-phosphate dehydrogenase. Clinical, genetic, biochemical aspects. A deficient G-6PD variant was discovered in 4 males of one family from northwestern Germany. Five generations of this family could be studied. The deficient G-6PD was a new variant, called "Gd (--) Aachen". Its main characteristics are the following: severe enzyme deficiency in erythrocytes (3% of normal), contrasting with an almost normal activity in leukocytes; normal molecular specific activity (i.e., normal ratio enzyme activity/cross-reacting material); slow mobility in starch gel electrophoresis (92-94% of normal); increased Michaelis constant for glucoes-6-phosphate (60-70 muM) and NADP+ (20-25 muM); decreased inhibition constant by NADPH with respect to NADP+ (7 muM); increased inhibition by ATP; normal utilization of the substrate analogues; slightly biphasic pH curve; thermal instability, and normal activation energy of the enzymatic reaction. The relationships between the hematologic disorders (severe and frequent hemolytic crises) and the unfavorable kinetic modifications are discussed.


The relation of sex, age, smoking status, birth rank and parental ages to pseudocholinesterase activity and phenotypes in a sample of Australian Caucasian adults. The frequencies of the pseudocholinesterase alleles E1u, E1a and E1f have been determined in a random sample of Australian residents. The frequency of E1a is the highest yet reported in a large caucasian sample. The considerable variation in E1f frequencies in previously reported samples is discussed in terms of possible sources of error in fluoride number determinations. Enzyme activity was found to increase with age in adulthood and was higher in males than in females. It was also positively correlated with dibucaine number in type U subjects. These observations are in conflict with those reported in previous investigations.


Confirmation of linkage between the loci for HL-A and glyoxalase I. Linkage analysis of 7 families with 30 children confirmed previous findings of the linkage between HL-A and GLO. Out of 28 families with 90 children the highest lod score value obtained was + 8.887 (recombination fraction = 0.1).


Increased sister chromatid exchange events in the human late replicating X. Human female blood cultures were labeled with BrdU for detecting sister chromatid exchanges (SCEs) by the Hoechst 33258 fluorescence technique. Late labeling with 3H-thymidine and autoradiography allowed the identification of the late replicating X. The mean number of SCEs in the cells was 13. The isopycnotic X showed an exchange frequency according to its relative length in the karyotype; in the late replicating X a doubled number of SCE events was observed.


Cell selection in vivo in normal/aneuploid chromosome abnormalities. Cytogenetic follow-up examination has been made of the 9 mixoploid children found among 11 148 newborn children. In 4 of the 9 children there was a significant increase in the frequency of the cell line with normal chromosome constitution and a significant decrease in the normal cell line was found in 1 child. In 4 there was no significant difference from the first to the last examination. The frequency of the cell line with normal chromosomes increased from 32-68% to between 93-97% in 3 cases and to 86% in 1. The possibility that children with mixoploid chromosome abnormalities at birth will reveal no cell line with chromosome abnormality in lymphocyte cultures as adults in spite of having clinical signs of the chromosome aberration found in one cell line at birth is discussed.


Tricho-rhino-phalangeal syndrome. The first case in Japan. The case of a 13-year-old girl with tricho-rhino-phalangeal syndrome is presented. It is characterized by sparse and slow growing hair, pear-shaped nose and cone-shaped epiphyses of hands and feet. The inheritance pattern is probably autosomal dominant transmission.


Pathomorphological changes in an early spontaneous abortus with triploidy (69, XXX). Triploid (69,XXX) spontaneous abortus with a gestational age of 14--15 weeks (anatomical age of the embryo was 6 weeks) was analyzed macro- and microscopically. There were hydatidiform swelling and cystic degeneration of the villi, without proliferatio of the trophoblast of cells, aplasia of one umbilical artery. The embryo had the following anomalies: cranial and caudal hypoplasia; aplasia of the facial structures (aprosopia), ocular vesicles, nasal stalk, extremity buds, somites, upper jaw, hyoid and pharyngeal arches, esophagus; trachea, Rathke's pouch and oropharyngeal cavity; encephalocele, focal anomalous rudiments of cartilage in the chordamesoderm, atresia of the stomodeal foramen and persistance of the lenticular placode.


Exomphalos and trisomy 18 syndrome. Report of two cases. A large exomphalose was found in two infants with a clinically and cytogenetically typical picture of trisomy 18 syndrome. In addition one infant was a case of male pseudohermaphroditism.


[A girl with a deletion (2) (q34q36): cytogenetic and clinical observations (author's transl)]. We describe a case of (2) (q34;q36) deletion in a girl. The main clinical features of the proband are staturo-ponderal retardation, skull-face dysmorphia and an interauricular communication. The relationship between morphological and chromosomal abnormalities is discussed.


Lymphocyte nucleoli activation as a marker of autoimmune disorder development. I. Observation in NZB mice. An increased number of lymphocytes with nucleoli synthesizing RNA was found in blood and lymph nodes of aged NZB mice who developed an autoimmune disease. This was not observed in young NZB mice and in 40NX mice where no immune disorders were noted.


Acquired resistance to ticks. II. Effects of Cyclophosphamide on resistance. Guinea-pig developed resistance to Dermacentor andersoni larvae after one infestation. Cyclophosphamide administered in one dose (300 mg/kg) 48 hr prior to an initial infestation with larvae blocked the acquisition of resistance. When cyclophosphamide was given in a similar regimen to guinea-pigs which had already acquired resistance, the expressin of resistance was partially blocked. It was proposed that the blockage of the acquisition of resistance further confirmed the immunological nature of tick resistance. Partial blockage of the expression of resistance by cyclophosphamide substantiated the presence of a humoral component to the resistance mechanism. The presence of a cell-mediated component was previously well established.


Subfragmentation of the Fc fragment of human IgG1 myeloma protein by thermolysin. Human IgG1 Fc fragment was digested at neutral pH by thermolysin, producing two large subfragments: one comprising the major part of the Fc fragment but devoid of the hinge region; the other comprising the Cgamma3 domain. The former fragment retained the capacity to react with "general" rheumatoid factors whereas the latter did not, indicating that the binding site for "general" rheumatoid factors on the Fc fragment of human IgG1 does not involve the hinge region of the molecule.


Inhibition of steroid induced gastric ulcers in forestomachectomized albino rats by phenobarbital. Oral administration of 2.5 mg of prednisolone/100 gm B.W. to starved forestomachectomized albino rats for three days induces 5-7 acute gastric ulcers with a severity of 3+ in 86% of the rats in the corpus comparable to human gastric ulcers with an ulcer index of 17.1. When 15 mg of Phenobarbital/100 gm B.W. was concomitantly administered with prednisolone either orally or subcutaneously prevents the formation of steroid induced ulcers probably due to their insensitization of gastric epithelium to corticoid ulcerogenic induction. This preliminary observation may have a therapeutic value as this sedative has a common usage. It is is suggested that forestomachectomized rats are best suited for ulcerogenic experiments.


A comparative study of a few tests of dynamic lung function. A variety of tests like FEV0-76, FEV1, PFR, MEFR is in use for assessment of ventilatory function of the lungs. Each of them has some marginal advantage over the other. It is, therefore, necessary to find out their relative merits and choose the one which can provide the maximum information in a reasonably short time. In this project, a norm of all the above tests for the people of Gujarat of age group 18-20 years has been found, the relative merit of the tests has been discussed and the velocity of air flow at 0.3 sec of expiration has been suggested as the single measurement which may conveniently replace all the other above.


A study of basal metabolic rate in adult Kashmiris. Sixty two adults, including forty one males (16-36 yrs.) and twenty one females (16-25 yrs.), were studied by closed circuit method as regards their B.M.R. Males showed a greater lowering of metabolism than the females when compared to MF and AD standards. The females actually showed a higher B.M.R. than the RR standards. Both the sexes had a higher basal metabolism than the value reported from other parts of the country. The difference, however, was significant only in the case of females. Winter metabolism was not sigificantly higher than the summer value in either sex, though females showed some what greater variation.


Mechanism of acti on of chronically administered cannabis extract on  the female genital tract of gerbils Meriones hurrianae. Daily administration of cannabis extract (2.5 mg/day for a period of 60 days) caused degenerative changes in the ovarian tissue. Luteinization was inhibited. Corpus-luteum degeneration was conspicuous. Distinct effects were produced upon the uterine biochemistry, consisting of decreased RNA, protein, sialic acid and glycogen concentration of the uterus. Vaginal RNA and protein contents were low. An anti-estrogenic action of cannabis extract in female gerbils is suggested.


Distinct deoxyribonucleic acid polymerase activities associated with Dane particles and naked Dane cores. Two distinct deoxyribonucleic acid polymerase activities were found associated with liver fractions containing Dane particles and Dane cores.


The role of imipramine hydrochloride on plasma transaminase and glucose level on rats under altitude stress. The effect of exposure of albino rats to simulated high altitude stress of 5000 metres for 3 and 6 hrs on plasma transaminases, non-protein nitrogen (NPN), total protein percentages and blood sugar levels have been studied both with and without the administration of an anti-stress drug Imipramine HCl at 2.0 mg/kg body weight. The drug appears to have a slight hepatotoxic effect.


Formation of cross-reacting antibodies against cellular and extracellular lipoteichoic acid of Streptococcus mutans BHT. Antisera prepared against strains of Streptococcus mutans, particularly strain BHT, contain antibodies to the membrane lipoteichoic acid component that will cross-react with lipoteichoic acids from three species of lactobacilli. A more detailed study of the antibodies to strain BHT confirmed that the antibodies are specific for the polyglycerol phosphate component common to the lipoteichoic acids. Sera with a higher hemagglutinating titer could be obtained by injecting disrupted organisms. The partially purified lipoteichoic acid isolated from the culture fluid of S. mutans BHT was also immunogenic antibodies; antibodies were also formed against another unidentified component in this extracellular fraction.


Increased fat degradation in the denervated muscle of frog. The total lipids triglycerides, free fatty acids, glycerol and acetoaectate levels in the gastrocnemius muscle of frog denervated for 1 month were compared. A significant atrophy without any change in total DNA content per whole muscle was noted on denervation. No significant change in the mitochondrial protein content in the atrophied muscle was noted. Relative to total muscle mass, the total fat, triglyceride, free fatty acids and glycerols decreased on denervation. This suggests the increased degradation of fat during denervation. Concomittantly lypolytic and esterase activities in the atrophied muscle increased. Palmitate and pyruvate oxidations in the mitochondria of atrophied muscle and also the enzymes of beta-oxidative scheme increased. 14C-acetate incorporation rates revealed that there is a decrease in fatty acid synthesis in the atrophied muscle. It was suggested that the increased fatty acid oxidation and decreased fatty acid synthesis in the atrophic process may not be the result of a simple functional demand; but may involve more factors in terms of neuro-muscular functions.


Chemical and biological properties of extracellular slime produced by Staphylococcus aureus grown in high-carbohydrate, high-salt medium. Slime material produced by three strains of Staphylococcus aureus grown in the high-carbohydrate, high-salt modified 110 medium contained ribitol teichoic acid and, in two of the three strains, a basic protein reacting with antisera to S. aureus whole cells and cell walls. The basic protein differed chemically and serologically from cell wall mucopeptide and protein A. Substances resembling the capsular antigen of the Smith diffuse strain of S. aureus were not detected, nor were any other uronic acid-containing components. When cell walls, slime material, and teichoic acid were injected intradermally into cows, only cell walls produced a skin reaction.


Subcutaneous multiplication of exfoliatin-producing staphylococci. After subcutaneous inoculation of approximately 10(6) cocci, changes in staphylococcal populations were followed by the enumeration of organisms in excised tissues. In contrast to conventionaal Staphylococcus aureus strains, exfoliatin-producing strains were able to multiply in the subcutaneous tissues of neonatal and adult mice. Although strains capable of producing large quantities of exfoliatin were better able to proliferate than strains producing lesser amounts of toxin, it was not determined whether exfoliatin was directly responsible for the observed multiplication. Two variants exhibiting a partial loss in exfoliatin production showed minor changes in proliferative capability. A third strain, after being cured of its exfoliatin plasmid, manifested a marked reduction in exfoliatin production and was unable to multiply subcutaneously. With some strains multiplication proceeded for several hours but was then followed by a decline in the number of viable organisms. Histological sections of subcutaneous lesions revealed a rapid influx of neutrophils, but leukocytes accumulated in the region regardless of whether the organisms multiplied or were eliminated.


Responses of mice immunized with influenza virus by serosol and parenteral routes. Antibody levels in sera and respiratory secretions and resistance to respiratory infections were examined in mice given live infuenza virus in small-particle (2 mum) aerosols, large-particle (10 mum) aerosols, intraperitoneally, and subcutaneously. After parenteral administration antibody was found primarily in the serum, but small amounts were recovered in bronchoalveolar washings after 2 to 3 weeks. Specific antibody was present in both sera and bronchoalveolar washings from mice given virus in small-particle aerosols to achieve virus dissemination throughout the respiratory tract. Immunoglobulin A, immunoglobulin G, and trace amounts of immunoglobulin M, all specific for the infecting virus, were detected in bronchoalveolar washings of small-particle aerosol-infected mice. Virus administration in large-particle aerosols (for primary virus localization in upper respiratory tract) at doses greater than those required to initiate infection with small-particle aerosols failed to stimulate production of antibody in sera or bronchoalveolar washings. Small-particle aerosol-immunized mice were resistant to subsequent challenge with 10(2.0) respiratory median lethal doses of virulent virus, whereas large-particle aerosol-immunized mice were not protected. Parenteral immunization modified the course of the disease in challenged mice and reduce mortality rates but did not prevent reinfection of the respiratory tract.


Synergistic protective effect in rabbits of immunization with Vibrio cholerae lipopolysaccharide and toxin/toxoid. Subcutaneous immunization of rabbits with a combination of Vibrio cholerae lipopolysaccharide (LPS) and enterotoxin induced a more than 100-fold-higher degree of protection against intestinal challenge with live cholera vibrios than did vaccination with either of the two antigens alone. Such a synergistic effect was also obtained by immunization with a combination of LPS and choleragenoid. The immunization with LPS and toxin (or toxoid) in combination did not enhance the reistance to toxin challenge above that induced by the toxin component alone. This, together with data from titrations of anti-LPS and antitoxin antibodies in serum and in intestinal washings, contradicts enhanced immune responses due to adjuvant action of the two antigens as the explanation for the synergistic effect of the combined vaccines. A more likely explanation would be that the antibacterial and antitoxic immune responses, without being increased in themselves, function synergistically by interfering with two separate events in cholera pathogensis.


Preparative isotachophoretic separation of skin test antigens from blastomycin purified derivative. This investigation examined the theoretical and practical parameters requisite to preparatory istachophoretic separation of blastomycin purified derivative. It resulted in a relatively simple, two-step procedure for preparation of blastomycin antigens in milligram quantitites that exhibited sensitivity and specificity in experimentally infected guinea pigs. Analysis of the nine isotachophoretic fractions for skin test sensitivity and specificity provided some insight into the generally accepted unreliability of blastomycin when used for immunological evaluation.


Effective immunity to dental caries: protection of malnourished rats by local injection of Streptococcus mutans. When rats from dams fed a low-protein diet were injected with whole, killed Strepococcus mutans 6715 cells in the region of the submandibular gland, they developed serum and salivary agglutinins to this microorganism. Titers of agglutinins in malnourished rats were similar to those observed in rats from dams fed a nutritionally adequate diet that were locally injected with S. mutans. Furthermore, both groups of immunized rats subsequently infected with cariogenic S. mutans 6715 had lower mean caries scores than infected, nonimmunized rats. This reduced incidence of caries scores than infected, nonimmunized rats. This reduced incidence of caries was evident on all molar surfaces. The mean body weights of immunized and nonimmunized, protein-deficient rats were not significantly different; likewise, both immunized and nonimmunize normally nourished rats exhibited similar weight gains. Malnourished rats, not immunized but infected with S. mutans, had significantly more caries than normal, nonimmunized infected rats. Both dietary groups of noninfected rats had very few carious lesions. These results suggest that carious lesions observed in these rats resulted from S. mutans 6715 infection. Furthermore, protein-malnourished rats, injected in the region of the submandibular gland with whole, killed S. mutans elicit an immune response and are protected against S. mutans-induced caries.


Aerosol vaccination of mice with a live, temperature- sensitive recombinant influenza virus. Mice were vaccinated intranasally (i.n.) or with small-particle aerosols (SPA; 2 mum) or large-particle aerosols (LPA; 8 mum) of an attenuated, temperature-sensitive, recombinant A influenza (H3N2) virus, ts-1 (E). Serum virus-neutralizing and hemagglutination inhibition antibodies were detected for all vaccinated mice by 28 days. Bronchoalveolar wash fluids had increased levels of immunoglobulin (IgG, IgA) only in the i.n. -vaccinated mice. Hemagglutination and virus-neutralizing antibodies were detected in the SPA- and i.n. -vaccinated groups but not in the LPA vaccinates. Upon challenge with SPA of a mouse virulent H3N2 influenza vitus, total protection was obtained for the SPA- and I.N. -vaccinated mice, whereas only 89% of the LPA group survived. Replication of the challenge virus was signifcantly repressed in both the lower and upper respiratory tracts of the three groups of vaccinated mice compared to the nonvaccinated controls. The protection afforded the SPA- and i.n. -vaccinated mice was the same as measured for mice after recovery from earlier subelthal active infection with virulent virus.


Host defenses in experimental scrub typhus: histopathological correlates. Intraperitoneal (i.p.) infection of BALB/c mice with 1,000 50% mouse lethal doses of the Karp strain of Rickettsia tsutsugamushi was inevitably lethal, and associated pathological alterations were confined to the peritoneal cavity. These included: (i) continuous proliferation of rickettsial organisms in peritoneal macrophages until death; (ii) hepatic granulomas appearing 6 days after infection and increasing in size and number until death; (iii) splenomegaly, resulting principally from proliferation of lymphoid tissue, and (iv) terminal peritonitis. Under two circumstances, i.p. infections with R. tsutsugamushi were not lethal: (i) infection with 100 50% mouse infectious doses of the Gilliam strain, which, in fact, resulted in immune protection against otherwise lethal Karp challenge; and (ii) Karp infection of animals immunized with the Gilliam strain. In both cases, the associated pathological abnormalities were, as with primary Karp infection, restricted to the peritoneal cavity. Also similar was the striking splenomegaly due to lymphoid proliferation, which was particularly prominent in immunized animals. In contrast to primary and lethal Karp infection, however, these infections were characterized by: (i) minimal and transient proliferation of rickettsial organisms in peritoneal macrophages; (ii) disappearance of hepatic granulomas; and (iii) absence of peritonitis. It was concluded that the survival of an animal bearing an i.p. infection of scrub typhus depended on its ability to concentrate a sufficiently vigorous immune response in the peritoneal cavity, resulting in the evolution of rickettsiacidal macrophages capable of suppressing the infection.


Cytotoxic effect of the thermostable direct hemolysin produced by Vibrio parahaemolyticus on FL cells. The thermostable direct hemolysin produced by Vibrio parahaemolyticus showed cytotoxic activity on FL cells derived from human amniotic membrane. Scanning electron micrographs of the whole cells showed that the microvilli on the cell surface decreased in number and changed in shape on treatment with the hemolysin. Most of the microvilli disappeared before death of the cells, as judged from the results of staining the cells with trypan blue and measuring release of alkaline phosphatase from the cells. Electron micrographs of thin sections ofthe cells showed that the cytoplasm of the cells was not significantly affected by treatment with sublethal amounts of hemolysin, even when the microvilli on the cell surface were significantly affected. Lethal amounts of hemolysin affected the cytoplasm and caused disarovilli on the cell surface are affected by treatment with the hemolysin before cytoxic effects develop.


Growth and effects of ureaplasmas (T mycoplasmas) in bovine oviductal organ cultures. Ureaplasmas isolated from the human genital tract and from the genital and respiratory tracts of cattle were grown in association with organ cultures of bovine oviduct (uterine tube). All strains of unreaplasmas multiplied in organ cultures, stopped ciliary activity, and caused histological lesions. Most strains grew well, and 10(8) to 10(9) color-changing units were determined 18 to 144 h after inoculation. Twenty-four to 144 h after inoculation with unreaplasmas, ciliostasis was complete. Ciliostasis was also caused by additions of nonviable cultures at pH 8.8 (or adjusted to 7.4) or washed disrupted cells (100 mug of protein/ml); it occurred in 48 to 96 h. The cilia-stopping effect of nonviable cultures was diminished by heating (56 C for 30 min) and was abolished by boiling. When added to fresh medium in amounts exceeding 25%, nonviable unreaplasmal cultures completely inhibited ureaplasmal growth. By light, scanning, and transmission electron microscopy, cilia-stopping effect was correlated with collapse and sloughing of the cilia (the initial lesion was "bent" cilia), with bulging and vacuolization of secretory and ciliated cells, and finally with disorganization of the epithelium, necrosis, and desquamation.


Unbalanced growth and macromolecular synthesis in Streptococcus mutans FA-1. The continued synthesis of deoxyribonucleic acid, protein, cell wall peptidoglycan and intracellular iodophilic polysaccharide (IPS) by Streptococcus mutans strain FA-1 after several treatments intended to inhibit protein synthesis was studied. Exponential-phase cultures were: (i) simultaneously deprived of two required amino acids (cystine and leucine) that are not present in the cell wall peptidoglycan of this species; (ii) depreived of required amino acids (lysine or glutamate plus glutamine ) that are present in both peptidoglycan and protein; or (iii) treated with tetracycline. Each of these three types of treatment was accompanied by a different pattern of unbalanced growth. The patterns of unbalanced growth that accompanied treatments (i) or (ii) differed substantially from the patterns observed previously for Streptococcus faecalis ATCC 9790, a noncariogenic organism that does not contain IPS. In contrast to S. faecalis 9790, S. mutans FA-1 failed to accumulate peptidoglycan and thicken its wall when deprived of non-wall amino acids. Instead, S. mutans FA-1 continued to accumulate IPS to levels substantially higher than those found in exponential-phase cells. Again, in contrast to S. faecalis, S. mutans FA-1 failed to autolyze upon deprivation of essential precursors of wall peptidoglycan. Under conditions of lysine of glutamate/glutamine deprivation, S. mutans FA-1 continued to accumulate IPS to very high levels. Treatment with tetracycline did result in peptidoglycan accumulation and wall thickening in a manner very similar to that observed previously for inhibition of protein synthesis in S. faecalis. Realtively little IPS synthesis continued after tetracycline treatment. Accumulation of IPS appeared to occur when both ribonucleic acid and peptidoglycan synthesis were severely inhibited. The observations are discussed in terms of the survival of cariogenic organisms in the oral environment.


Lymphocytic choriomeningitis virus infection in fetal, newborn, and young adult Syrian hamsters (Mesocricetus auratus). The pathogenesis of lymphocytic choriomeningitis virus infection in fetal, newborn, and young adult hamsters was studied. Infected newborn hamsters initially developed a persistent viremia and viruria with titers often in excess of 10(4.0) mean infectious doses/0.03 ml of blood or urine. After week 12 two different patterns of infection became evident. Approximately one-half of the hamsters eventually cleared the infection, whereas the others developed a chronic progressive and ultimalely fatal disease characterized by continuous high-titered viremia and viruria and high titers of virus in their tissues. Complement-fixing antibody and, to a lesser degree, virus-neutralizing antibody coexisted with the viremia. Hamsters with persistently high levels of viremia and viruria developed chronic glomerulonephritis and widespread vasculitis, whereas hamsters that cleared their infections did not develop these lesions. Litters of hamsters born to viremic mothers were invariably infected. Litter sizes were small and breeding effectiveness was reduce; however, vertical, congenital infection was successfully passed through three generations. The course of infection in the congenitally infected hamsters was similar to that in newborn infected hamsters, with all animals producing complement-fixing antibody, some animals being capable of clearing the viremia and remaining healthy, and other animals having persistent viremia and fatal disease. Inoculated young adult hamsters did not become diseased, developed viremia and viruria which persisted up to 3 and 6 months, respectively, and developed complement-fixing antibody by 10 days after infection. The prolonged urinary excretion of large amounts of lymphocytic choriomeningitis virus by asymptomatic, chronically infected hamsters is an important public health consideration when dealing with potential human infection.


Purification of staphylococcal alpha-toxin by adsorption chromatography on glass. Staphylococcal alpha-toxin was purified from Staphylococcus aureus growth medium using adsorption chromatography on controlled pore glass beads. Elution of alpha-toxin from the unmodified glass surface of the beads with various anions generally followed the chaotropic series. Alpha-toxin, purified by glass bead chromatography, is composed of a single electrophoretic form, containing less than 2% of other forms.


Inhibition of lethality in endotoxin-challenged mice treated with zinc chloride. Zinc chloride protected against lethality in mice undergoing endotoxin shock.


Solid-phase radioimmunoassay for antibodies to flavivirus structural and nonstructural proteins. A micro-solid-phase radioimmunoassay (SPRIA) is described for quantitation of antibodies to purified flaviviruses as well as to the purified envelope glycoprotein and 80,000-molecular-weight viral nonstructural protein. Sera from mice experimentally infected with Saint Louis encephalitis (SLE) virus or from humans after a primary SLE virus infection reacted more specifically with the major viral envelope protein in the SPRIA test than with antigens conventionally used in the complement fixation (CF) and hemagglutination inhibition tests. A high degree of correlation (P is less than 0.05) was observed between SPRIA anti-immunoglobulin G binding values with the 80,000-molecular-weight nonstructural protein of SLE virus and antibody titers obtained by plaque reduction neutralization and CF with the nonstructural protein. In five of seven human sera in which CF antibody titers to the nonstructural protein were 4 or less, SPRIA testing revealed significant titers of IgG immunoglobulin reactive with this viral protein. The SPRIA test for antibodies reactive with group B togavirus nonstructural protein is as specific and sensitive as the plaque reduction neutralization test for titrating viral antibody in human and animal sera. Antibodies reactive with viral envelope proteins are broadly cross-reactive by the Spria technique, demonstrating both group- and complex-reactive antigenic determinants. The SPRIA test, using wells precoated with antigen, can be completed in 1 day, providing a rapid, highly sensitive test which can be adapted to use in testing a large number of sera.


Indirect blastogenesis of peripheral blood leukocytes in experimental gingivitis. The blastogenic response of peripheral blood leukocytes to lipopolysaccharide (LPS) was followed over a short course of experimental gingivitis, developed in human volunteers who strictly avoided oral hygeine procedures for periods up to 9 days. Eleven young males initially received thorough dental prophylaxes and supervised oral hygeine until they acquired optimal gingival health. At this point, leukocytes (5 X 10(5)) incubated with 1.5 to 25 mug of LPS in serum-free media showed no response as measured by tritiated thymidine uptake. Coincubation of cells with LPS and phytohemagglutinin (PHA), however, caused synergistic enhancement of blastogenesis in every LPS-PHA dose combination tried. With progressive accumulation of dental plaque and the concomitant development of gingival inflammation, this synergistic response was lost and replaced, proportionately, by a direct response to LPS. The leukocyte response to PHA was marginally enhanced with gingivitis.


Importance of Actinomyces and certain gram-negative anaerobic organisms in the transformation of lymphocytes from patients with periodontal disease. Dental plaque deposits are known to be potent stimulants of lymphocyte transformation in patients with periodontal disease but not in normal subjects. Since plaque deposits consist mainly of whole bacteria, the cell walls of the most commonly found organisms in plaque were tested for their capacity to induce lymphocyte transformation. There was a direct correlation between the severity of peridontal disease and the amount of transformation induced by the cell walls of oral bacteria and by solubilized dental plaque. Cord blood leukocytes and lymphocytes from clinically normal people did not respond, which indicates that these stimulants are antigens rather than mitogens. Of the eleven bacteria tested, four members of the family Actinomycetaceae (Actinomyces viscosus, A. israelii, A. naeslundii, and Arachnia propionica), the related Propionibacterium acnes, and an anaerobic gram-negative anaerobic rod (27N). The high prevalence of the former organisms in the mature dental plaque that forms around the gingival crevice area and the potent efficacy with which they stimulate lymphocytes indicates that Actinomyces and certain gram-negative anaerobes may be important etiological agents in chronic periodontal inflammation in man.


Modification of resistance of mice to Naegleria fowleri infections. Naegleria fowleri, which produces a fatal meningoencephalitis in humans, is also able to produce a progressive and fatal disease in mice. The course of the disease in DUB/ICR mice is dependent upon the infecting dose of organisms, whether administered intraperitoneally (i.p.) or intravenously (i.v.). All of the mice receiving 10(7) trophozoites/mouse i.v. or 4.85 X 10(7) trophozoites/mouse i.p. were killed within 10 days. Escherichia coli O26:B6 lipopolysaccharide, administered at a dose of 1 mg/kg 24 h prior to N. fowleri, afforded some protection for several days after challenge, but by day 8 there was no difference in survival of untreated and endotoxin-treated mice. No significant protection was afforded by a complex of lipid A with concanavalin A (ConA) or bovine serum albumin (BSA) or by dimethylmyristamide-BSA, dimethylmyristamide, BSA, beta-hydroxymyristic acid-ConA, beta-hydroxymyristic acid, ConA, myristic acid-BSA, or myristic acid. Mice surviving primary i.v. or i.p. challenge doses of N. fowleri, 5 X 10(6) and 10(7) trophozoites/mouse, respectively, were highly resistant to rechallenge with an i.v. dose of organisms (5 X 10(6) Naegleria/mouse) that produced uniformly fatal disease in untreated control mice.


Maturation of intestinal defenses against peroral infection with group B coxsackievirus in mice. The intestinal tract of adult mice provides effective protection against peroral infection with group B coxsackievirus. This protective function consists of at least two separate components. One is a barrier effect that prevents virus from passing through the mucosal side of the gut into the circulation. It becomes clearly evident at 18 days of life and is present thereafter. The other is a clearance mechanism that acts to eliminate virus from the enteric tract after infection has occurred. This is first demonstrable at about 14 to 18 days and also persists. The appearance of these protective functions coincides with the known development of enzymatic and morphological changes in the gut associated with the transition from suckling to weanling.


Temperature-sensitive mutants of rabies virus in mice: a mutant (ts2) revertant mixture selectively pathogenic by the peripheral route of inoculation. Analysis of the pathogenic potential in mice of a variety of rabies and rabies serogroup viruses revealed that an apparently revertant population of virus derived from CVS mutant ts 2 had a unique capacity to selectively induce paralytic disease when given by a peripheral [intraplantar (i.pl.)] route of inoculation. Little paralytic disease was induced by high concentrations of virus administered by the intracerebral (i.c.) route, whereas paralytic disease and death were characteristically induced in mice given only a few infectious doses of virus i.c. Disease induced by i.pl. inoculation was dose dependent. Mice frequently survived paralytic disease induced by i. pl. inoculation, with clinical signs often persisting indefinitely; mice surviving i.c. inoculation of high concentrations of virus frequently exhibited chronic nonspecific signs of minor debility. Analysis of the ts 2 virus population indicated that it was composed of a mixture of ts and revertant virions, each with characteristic pathogenic (or nonpatholgnic) propensities, none of which was identical to the original composite ts 2 virus populations. Despite the heterogeneity of the ts 2 virus population, the typical pathogenic pattern of selective pathogenic capicity after i. pl. inoculation at high doses was retained during 11 ocnsecutive passages in suckling mouse brain. ts 2 virus was demonstrated to interfere with the disease-producing capacity of CVS fixed rabies virus when ts 2-CVS mixtures were inoculated i.c. However, attempts to demonstrate a particular propensity for induction in vitro of "autointerference" by ts 2 in serial passage in BHK-21 cell culture inoculated at high multiplicity were unsuccessful.


Interaction of diphtheria toxin and its active subunit, fragment A, with toxin-sensitive and toxin-resistant cells. Diphtheria toxin and purified fragment A, the active subunit of the toxin, were tested on toxin-sensitive and permeability class toxin-resistant cultured mammalian cells. Protein synthesis was inhibited to the same degree in sensitive or resistant cells by active concentrations of purified fragment A. In contrast, resistant cells required a concentration of whole toxin 5 to 6 logs greater than that required by sensitive cells to achieve the same degree of inhibition. On a molar basis, the toxicity of fragment A was equivalent to that of whole toxin on resistant cells. These results are evidence for the existence of two independent mechanisms for the entry of toxin or its active moiety into cells. One mechanism is a highly efficient, toxin-specific entry mechanism, involving surface receptor and fragment B-mediated association, and is active in sensitive cells. The other is a less efficient, nonspecific mechanism, probably related to endocytosis, which is operative in sensitive and resistant cells but is inapparent in sensitive cells when they are exposed to whole toxin because of the action of the specific mechanism.


Antibody-dependent cell-mediated cytotoxicity in cows: comparison of effector cell activity against heterologous erthrocyte and herpesvirus-infected bovine target cells. Bovine peripheral blood leukocytes (PBL) and cells collected from the bovine mammary gland were assayed for antibody-dependent cell-mediated cytotoxicity (ADCC) against chicken erythrocyte (CRBC) and bovine herpesvirus-infected bovine kidney cell targets. Bovine antisera were used to sensitize target cells. Both PBL and mammary leukocytes expressed ADCC, with the latter cell population having greater activity against both target cells. Only the CRBC target cells were killed by nonadherent PBL and phagocyte-depleted PBL. Nonadherent mammary leukocytes, rich in monocytes and macrophages, did kill virus-infected target cells. Carbonyl iron-treated mammary leukocytes failed to kill virus-infected targets but could destroy CRBC targets. Antimacrophage serum inhibited lysis of both CRBC and virus-infected targets, but antilymphocyte serum only inhibited CRBC killing. These observations indicated that at least two kinds of cells could mediate ADCC against CRBC but only cells of the mononuclear phagocytic series could kill virus-infected target cells. The herpesvirus-infected target cells became susceptible to ADCC 9 h after virus infection. A case is made for investigating the phenomenon of ADCC using in vitro systems that closely mimic the in vivo situation. The possible role of the ADCC mechanism as instrumental in causing recovery from herpesvirus infections is discussed.


Different effects of phytohemagglutinin-activated lymphocytes and their culture supernatants on macrophage function. When rabbit peritoneal exudate cells were incubated for 24 and 48 h with phytohemagglutinin-activated lymphocytes or their culture supernatants, two times as many cells remained adherent to culture slides as in the controls. More spreading cells were found among the adherent cells in the stimulated cultures. Eighty percent of spreading cells that were induced by supernatants were negative or faintly positive for beta-galactosidase. On the other hand, half of the spreading cells induced by activated lymphocytes were positive (1+ to 4+) for beta-lymphocytes and their supernatants. Under similar conditions, unstimulated peritoneal cells showed less marked activation. These findings show that macrophages can appear morphologically activated and yet not be enzymatically activated by lymphokines. Possible mechanisms of direct interaction of activated lymphocytes and macrophages are discussed.


Fusobacterium necrophorum infection in mice as a model for the study of liver abscess formation and induction of immunity. A mouse model is described in which intraperitoneal injection of Fusobacterium necrophorum results in chronic liver abscesses. Viable bacterial counts from mouse lung, liver, and spleen were obtained after whole organ homogenization. From 2 h to 5 days postchallenge, liver was found to contain more bacteria than lung on a per gram basis. Bacterial counts from liver and spleen were about the same during the first 8 h; thereafter liver was found to contain more bacteria. By day 13, though bacterial counts were equivalent in the three organs, abscesses were only observed in liver. This predilection for the liver may be due to a nutritional and/or microenvironmental factor(s). Blood cultures of infected mice revealed a general lack of bacteremia. Extended immunization with formalin-killed cells was found to protect mice against F. necrophorum infection.


Morphological and steroidogenic changes in cultured adrenal tumor cells induced by a subunit of cholera enterotoxin. A purified subunit of the cholera enterotoxin molecule was found to have morphological and steroidogenic inducing effects similar to those induced by the native enterotoxin on monolayer tissue cultures of Y1 adrenal tumor cells, although 1,000 times more subunit than toxin (weight basis) was required for maximal effects. In contrast to the whole toxin, the effects of the active subunit could not be prevented by prior incubation with either Gm1 ganglioside or with antibodies directed against choleragenoid (the binding subunit). These results suggest that different receptor sites may exist on cells for the binding and for the active subunits of cholera enterotoxin and/or that the active toxin fragment may exert its effects after gaining access to the intracellular compartment.


Streptococcus mutans dextransucrase: availability of disaggregated enzyme after growth in a chemically defined medium. The soluble dextransucrase (EC 2.4.1.5) activity produced by Streptococcus mutans strain 6715 during growth on a chemically defined synthetic medium (FMS) was compared to enzyme from glucose broth cultures (TSB). Growth on the two media was similar. The specific activity of ammonium sulfate-precipitated FMC enzyme was 17 times greater than similar TSB enzyme preparations. The FMC enzyme was stimulated 11-fold, whereas the TSB enzyme was stimulated 1.2-fold by the addition of exogenous primer dextran. In contrast to the TSB enzyme, the FMC activity could be disaggregated to a low-molecular-weight form by 1 M salt. Thus, low-molecular-weight S. mutans dextransucrase activity free of contaminating primer glucan may be readily obtained after growth of the bacterium in a chemically defined sucrose-free medium.


Immunoglobulin-bearing cells in lungs of mice infected with influenza virus. In sequential studies of cellular infiltrates from influenza-infected mouse lungs, increased populations of lymphocytes with surface immunoglobulins were observed, and immunoglobulin A-bearing cells exhibited the greatest relative increase.


Strain-dependent differences in murine susceptibility to toxoplasma. Susceptibility of seven inbred and outbred strains of mice to infection with trophozoites of Toxoplasma gondii at two different doses was determined. Stroking strain differences in susceptibility and changes of susceptibility with dosage change were seen. The data are consistent with the hypothesis that this may be due to genetic factors.


Studies on allergens of Dermatophagoides pteronyssinus by direct and indirect RAST. Dermatophagoides pteronyssinus (DP) extract was fractionated by Sephadex G-75 and liquid isoelectric focusing (IEF) and the allergenic activity of different fractions was monitored by direct and indirect RAST. The fractionation on Sephadex G-75 showed that the allergenic activity of DP extract was related to wide molecular weight spectrum components, even though the maximum amount was recovered in effluent that contained protein with a molecular weight ranging between 25,000 and 12,500 daltons. By fractionation of the mite extract on IEF, three main peaks of allergenic activity (pI less than 3.0; pI = 4.3 +/- 0.25; pI = 6.4 +/- 0.25) were found. Cross-inhibition experiments showed a high degree of cross-reactivity between allergenic material eluted in very distant regions of molecular weight or isoelectric point. The allergenic activity of unfractionated mite extract and of its IEF fractions was destroyed by pronase - but not by neuraminidase - treatment. These results suggest that DP extract probably contains one main allergen existing in multiple molecular forms rather than several distinct allergens and that a protein moiety of the allergen is necessary for the combination with IgE.


Effect of insulin and alloxan diabetes on carrageenin inflammation in rats. Some hypersensitivity reactions and allergic responses are known to be increased by insulin treatment and decreased in diabetes. In contrast, the present experiments showed that paw swelling induced by carrageenin in rats was inhibited by insulin. The anti-inflammatory activity, to some extent, paralleled the dose applied and did not appear to be due to hypoglycaemia. Alloxan diabetes in turn increased the vascular response to carrageenin and abolished the anti-inflammatory effect of insulin. The experiments call attention to the possible significance of insulin in the regulation of the acute non-immune inflammatory process.


Anaphylactoid reactions in connection with infusion of invert sugar solutions are due to macromolecular contaminants. 13 untoward mold anaphylactoid reactions were observed in patients infused with invert sugar solutions in Sweden during a 7-month period in 1973: an incidence of 1/31,000 infusions. Immunological and physicochemical analysis of invert sugar solutions and of the raw material, sucrose, revealed traces of native alpha-1,6-glucan with molecular weight of 10-100 millions as contaminant. This indicates, that the sucrose had been exposed to microbial contamination during its manufacture from sugar-beet or sugar-cane. Reversed single radial immunodiffusion was used for alpha-1,6-glucan detection and screening of all sucrose batches. Rejection of contaminated sucrose as raw material reduced the incidence of anaphylactoid invert sugar reactions to about 1/575,000 infusions. Examination of all sucrose raw material for traces of crude alpha-1,6-glucan is recommended as a test for detection of microbial contamination.


Lymphocyte stimulation and plasma inhibition in patients with malignant neoplasms. A comparative study with three mitogens. The blastogenic response of lymphocytes from patients with malignant neoplasms was evaluated by stimulation with three phytomitogens (PHA, PWM, and Con A). The response of patient lymphocytes to all three mitogens was significantly lower than that of control lymphocytes, and most patients with abnormal PHA responses also responded abnormally to PWM and Con A. However, a few patients with normal PHA responses were abnormal to Con A, suggesting the suppression of a Con A-sensitive population. The observation that PWM responses were abnormal in patients with lowered PHA lymphocyte stimulation indicates that both T and B lymphocyte mitogen responses were suppressed in these patients. Plasma from patients was capable of either inhibiting or enhancing lymphocyte mitogen stimulation. However, inhibitory plasmas were generally from patients with abnormal mitogen responses.


Modulation of the release of SRS-A from bovine lung in vitro by several autonomic and autacoid agents. The immunological release of slow-reacting substance of anaphylaxis (SRS-A) from bovine lung is modulated by several autonomic and autacoid agents. Isoproterenol, a beta-adrenergic agonist, inhibited release of SRS-A, as did cyclic AMP. Phenylephrine also inhibited SRS-A release but  through alpha-adrenergic mechanisms. This is in contrast to observations in human lung. Carbachol enhanced the release of bovine SRS-A. Histamine inhibited SRS-A release through the H2-receptor. Dopamine enhanced the release of SRS-A by means of a dopaminergic receptor. 5-HT did not significantly modulate SRS-A release.


Allergic contact dermatitis to low molecular weight allergens in the chicken. We have produced allergic contact dermatitis (ACD) to low molecular weight allergens in the chicken. Sensitization was induced by the application of a high concentration of allergen to a site on the comb previously treated with dimethyl sulfoxide. Challenge, which was made on the wattle with a nonirritating concentration of allergen, resulted in a dermatitis that was grossly and histologically similar to ACD in mammals. Chickens sensitized to oxazolone reacted to that allergen and not to picryl chloride, and conversely. This model provides a unique system wherein to study the T-cell and B-cell contributions to the generation, maintainance and expression of the ACD response.


Stabilisation, co-ordination and structuralisation in the integrated automation of complex systems. The objectives of structural parallelism and composition of modules in integrated automation, as well as in the tracking approach based on the distance between the systems, form the base of a methodology of analysis and synthesis of the control of complex systems. The unit control is founded on the tracking of trajectories determined by the behaviour of the whole complex global system. The control system is based on the desire to attain the imposed objectives, as well as a participation in the efforts of other systems. These properties of each decision centre thus formed, permit the stabilisation of the whole interconnected global system. This global system can easily be co-ordinated or restructured by the directing decision centre which disposed of a global model.


The derivation of second degree rate equations arising from two-substrate, two-product enzyme catalysed reactions whose catalytic cycle is branched. A selection of two-substrate, two-product enzyme catalysed reactions with alternate catalytic cycles and second degree rate equations was examined by the computer program described by Kinderlerer and Ainsworth (1976). It is shown that the numerator terms of the rate equations are particularly simple, easy to derive and capable of creating a broad division of the mechanisms considered. THE NUMERATOR TERMS ALSO HAVE THE ADVANTAGE OF BEING UNAFFECTED BY DEAD END INHIBITION OR THE PRESENCE OF Theorell-Chance reactions. It is then shown that the relatively small group of mechanisms, isolated by the numerator analysis, can be distinguished from each other by considering the denominator terms of the rate equation that are second degree in the concentration of the varied substrate.


Programming for the inexperienced users: machine independence and free-format input. Two programming techniques which can make the use of computers less troublesome to inexperienced users are described. The first has the object of making programs readily transferable from one machine to another, thereby facilitating transfer of programs between individuals. The second consists of a FORTRAN subroutine, described in detail, which allows numbers to be read by a program without being confined to specific columns on a punched card, thereby greatly simplifying data preparation.


Convolution computer processing of the brain electriacl image transmission. A propagation model of electrical activities between the cerebral cortex and the scalp is presented in this paper. This work is based on a hypothesis of a distribution of cortical sources in a non-uniform dipolar sheet. The general field equations show that the electrical image of the cortex on the scalp is given by the convolution of the cortical potential and a transfer function. We then present several numerical evaluations with respect to the attenuation of the signal, the visual field of the electrodes and the approximation errors that have been introduced. Simulation experiments then enable us to test the behaviour of the model in comparison with complex electrical activities.


Computer-aided investigations of respiratory data. In the first part of this paper a computerised evaluation of plethysmographic signals is presented. The off-line processing described eliminates some erroneous simplifications that routine plethysmography is bound to involve. (1) A subtraction of the box pressure baseline permits measurements with perfectly sealed constant volume plethysmographs even in the case of thermal disequilibrium. (2) Alveolar pressure is computed from the box pressure signal, using instantaneous values of the intrathoracic gas volume. (3) This technique enables one to utilise all the measured data for determinations of airway resistance and viscous work, thus yielding more exact values as compared to the graphic evaluations. In Part 2 a three-parameter model of transpulmonary mechanics, applied to fit data of cat experiments, is described. The aim of this study is to demonstrate how transpulmonary parameters (static pleural pressure, compliance and resistance) are modified by the actual state of the pulmonary vascular bed.


Detection of tumor-associated antigen in human melanoma cell line supernatants. Spent tissue culture medium (CDM-S) removed from a single cell line of human malignant melanoma grown in serum-free CDM, contained tumor-associated antigenic activity. Antibodies to CDM-S measured by complement fixation were detected in 44% (31/70) melanoma, 55% (15/27) sarcoma, 63% (24/38) carcinoma and 15% (11/72) normal sera. Delayed cutaneous hypersensitivity reactions (DCHR) were demonstrated in 4/5 melanoma patients at a 500 mug dose, 3/5 at a 100 mug dose and in 1/7 carcinoma patients at the 500 mug dose. One ml of CDM-S was shown to contain antigen equivalent to that obtained from the membranes of 2.9 X 10(7) tissue-cultured melanoma cells. After purification, 84% (16/19) sera from melanoma patients, 66% (12/18) from sarcoma and carcinoma patients and 8% (2/26) from normal controls were positive to the antigen by complement fixation.


Assessment of cell-mediated immunity to malignant mesothelioma by microcytotoxicity tests. Cell cultures were established from pleural effusions of patients with pleural mesothelioma, and peripheral mononuclear effector cells were tested for cytotoxicity against these cells by means of microcytotoxicity assay. Effector cells were obtained from normal healthy donors and from persons exposed occupationally to asbestos, including apparently healthy persons, patients with benign pleural conditions and patients with malignant mesothelioma. The overall incidence of cytotoxicity was low, and there was no evidence of increased cytotoxicity in mesothelioma patients or other asbestos-exposed donors. It is concluded that little or no tumour-directed cell-mediated immunity is detectable against malignant mesothelioma by microcytotoxicity methods.


Mortality from cancer of the uterus in Canada and its relationship to screening for cancer of the cervix. Age-standardized death rates at ages 30-64 from cancer of the uterus in the 10 provinces of Canada have been assessed in 1950-52, 1960-62 and 1970-72, and changes in rats over these periods related to screening for cancer of the cervix and variables derived from the censuses of 1961 and 1971. Death rates at the county and census division level have also been assessed for the provinces of Nova Scotia, Quebec, Ontario, Manitoba and Alberta for 1960-62 and 1970-72 and related to screening and census-derived variables. There is little indication of a contribution of screening to the fall in death rates that occurred in eight of the 10 provinces from 1950-52 to 1960-62. There is, however, evidence of an important and significant contribution of the intensity of screening (as indicated by the number of cervical cytology examinations conducted in 1966 expressed as a rate per 1,000 female population aged 20 or more) to the fall in death rates at both the provincial and county or census division levels from 1960-62 to 1970-72, and this effect is not abolished by taking the census-derived variables into account. It is concluded that a significant effect of intensity of screening on the reduction in mortality from cancer of the uterus in Canada in the age-group 30-64 over the period 1960-62 to 1970-72 has been demonstrated.


Rapid development of hyperplastic nodules and cirrhosis in the liver of rats treated concurrently with thioacetamide and the pyrrolizidine alkaloid lasiocarpine. Pyrrolizidine alkaloids have long been considered to be hepatotoxic in man as well as in grazing animals. To investigate the effect of liver cell division induced by thioacetamide on the hepatic changes induced by these alkaloids, rats were treated concurrently with thioacetamide and the pyrrolizidine alkaloid lasiocarpine. Thioacetamide was given intraperitoneally in a dose of 50 mg/kg b. wt twice weekly and lasiocarpine was administered in the diet at a concentration of 50 ppm. At 15 weeks, the combination of thioacetamide and lasiocarpine produced numerous grossly visible grey nodules in livers of 26 of 30 rats. Microscopically, these livers revealed a severe degree of postnecrotic cirrhosis and numerous hyperplastic nodules. The cells in most nodules were arranged in solid sheets or in a trabecular pattern and shown atypia, mitosis and hyperchromasia. In contrast, there was no evidence of cirrhosis or nodule formation in livers of animals treated with either lasiocarpine or thioacetamide alone. The rapid development of liver lesions in rats treated simultaneously with low doses of lasiocarpine and thioacetamide suggests that cell proliferation accentuates the development of neoplasia.ł


Interaction of antibody and cell surface localized antigen. The changes occurring in the expression of a membrane-associated antigen following binding of specific antibodies and the fate of the bound antibodies were investigated. Daudi cells, carrying membrane 7S IgM molecules, were coated with radio-labelled anti-IgM antibodies and incubated under culture conditions at 37degrees C. Within 10 h, more than 80% of the cellbound radioactivity was shed from the cells into the culture medium. Concomitantly, IgM molecules were lost, as the cells had a diminished ability to bind a new aliquot of antibody. This phenomenon occurred also when the antibody was used at a low concentration, i.e. did not saturate the cell-surface antigens. The precipitability of the antibody released from the cells with 10% TCA and 40% saturated ammonium sulphate was lower than that of native antibody, indicating some degradation. Also, shed antibody could rebind to fresh Daudi cells less efficiently than native antibody. The presence of immune complexes in the culture supernatants was indicated by the following results: (1) some of the shed radioactivity could bind to IgM-negative but Fc receptor-bearing cells; and (2) immune precipitates of sheep anti-IgM shed from 3H-leucine labelled Daudi cells and of rabbit anti-sheep IgG antibodies contained 3H activity.


Interferon and murine leukemia. VII. Therapeutic effect of interferon preparations after diagnosis of lymphoma in AKR mice. Daily administration of potent mouse interferon preparations, begun after clinical diagnosis of lymphoma in AKR mice, increased average survival by approximately 100%. Interferon treatment delayed the evolution of the lymphoma but regression of tumour was not observed. The therapeutic effects observed with interferon in AKR mice compare favorably with reported results obtained using standard anti-cancer drugs.


Modification of the effect of C. parvum on macrophage activity and tumour growth by X-irradiation. The radiosensitivity of three forms of response to injection of C. parvum in mice has been investigated. The increase in phagocytin index evoked by IP or IV injection of 0.7 mg C. parvum (but not that evoked by 1.4 mg) was reduced but not abolished in mice given 350-500 rad whole-body irradiation 4 days before C. parvum injection. Irradiation (500-1,000 rad) 4 days after C. parvum injection had no such effect. The antitumour cytotoxicity in vitro of PE and spleen cells from C. parvum-treated mice was abolished by irradiation of the cell donor (400-800 rad) 4 days before C. parvum injection, but was not reduced by irradiation (800 rad) of the cell donor 4 days after C. parvum injection or of the effector cells in vitro. The antitumour response to systemic (IP) injection of C. parvum was reduced by 350-500 rad whole-body irradiation, irrespective of whether this was given 4 days before or 4 days after C. parvum injection. The response to intratumour injection of C. parvum was even more radiosensitive. It has been suggested in previous papers from this laboratory that the antitumour effect of C. parvum is related to its capacity to stimulate macrophage activity, although in addition T lymphocytes are necessary for local injection to be effective and non-T lymphocytes may be concerned in the response to both local and systemic injection. The present results in no way conflict with this view. They suggest in addition that the effect of C. parvum on the macrophage system is, to a considerable extent, due to stimulation of macrophage precursors to differentiate into actively phagocytic and cytotoxic mature cells.


The effect of BCG on extravascular mononuclear cell accumulation in vivo. BCG injection produces an increased accumulation of adoptively transferred 51Cr-labelled mononuclear cells at the site of inflammatory reactions in rats by two mechanisms. Systemic immunization induced a population of "activated" cells which, compared with controls, had an increased life span in the circulation after transfer and migrated in increased numbers to a chemotactic stimulus. These cells also were found in significantly greater than normal levels in the liver. In addition, local injection of BCG selectively attracted adoptively transferred mononuclear cells. These mechanisms help to explain local mononuclear cell accumulation which accompanies tumor rejection following intralesional or systemic BCG immunization.


Studies on trypsin inhibitors. Part V. Synthesis of the protected octapeptide (sequence 36-43) of porcine pancreatic secretory trypsin inhibitor II (Kazal). Synthesis is described of the partially protected octapeptide tert-butyloxycarbonyl-gamma-tert-butylglutamylasparaginyl-N-trifluoroacetyllysyl-N-trifluoracetyllysylarginyl-Ngamma-4,4'-dimethoxybenzhydryglutaminylthreonylproline  corresponding to positions 36-43 of the amino acid sequence of porcine pancreatic secretory trypsin inhibitor II. The tetrapeptide free base arginyl-Ngamma-4,4'-dimethoxybenzhydrylglutaminylthreonylproline was acylated, by the azide proceedure, with the tripeptide benzyloxycarbonyl-asparaginyl-N-trifluoroacetyllsyl-N-trifluoroacetyllysine hydrazide. The resulting protected heptapeptide was partially deblocked by catalytic hydrogenation and reacted with alpha-1-succinimidyl-gamma-tert-butyl tert-butyloxycarbonylglutamate. The stereochemical homogeneity of the ensuing octapeptide was assessed, after partial deprotection with aqueous 90% trifluoroacetic acid, by digestion with papain and aminopeptidase M followed by quantitative amino acid analysis.


Studies on trypsin inhibitors. Part VI. Synthesis of the protected nonapeptide (sequence 44-52) of porcine pancreatic secretory trypsin inhibitor II (Kazal). Synthesis is described of the partially protected nonapeptide tert-butyloxycarbonyl-valylleucylisoleucylglutaminyl-N-trifluoroacetyllsylserylglycylprolyl-S-acetamido-methylcysteine corresponding to positions 44-52 of the amino acid sequence of porcine pancreatic secretory trypsin inhibitor II. The hexapeptide free base glutaminyl-N-trifluoroacetyllsylserylglycylprolyl-S-acetamidomethylcysteine, prepared by two alternative routes, was acylated by the azide procedure, with the tripeptide tert-butyloxycarbonylvalylleucylisoleucine hydrazide. The stereochemical homogeneity of the resulting nonapeptide was assessed, after partial deprotection by treatment with aqueous 90% trifluoroacetic acid, by digestion with papain and aminopeptidase M followed by quantitative amino acid analysis.


Non-random relationships among amino acids in protein sequences. We have computed the observed distribution of amino acid pairs n, n+1 up to n, n+10 within 100 unrelated protein sequences containing a total of 14,034 amino acids. The expected distribution of the same amino acid pairs was calculated assuming that the amino acids within each sequence are arranged in random order. Statistical analysis of the data shows that amino acids in the n, n+1 and n, n+3 relationship deviate from random expectation at the 1% level of significance. These relationships are consistent with known secondary structures; however, there are no amino acid pairs which obviously contribute to the non-random result.


On the reaction of acetamidomethanol with native and reduced bovine pancreatic trypsin inhibtor (Kunitz inhibitor). The stability of native and reduced bovine pancreatic trypsin inhibitor (Kunitz inhibitor) in anhydrous hydrogen fluoride and their reaction with acetamidomethanol, in the same solvent, have been investigated. The bovine Kunitz inhibitor appears to be stable in liquid hydrogen fluoride but the reduced molecule loses about 50% of its ability to regain inhibitory power, upon air oxidation, by exposure to this solvent. Tyrosine residues appear to be affected by acetamidomethylation of the native protein to give a modified inhibitor which is still highly active in inhibiting trypsin. The extent of correct refolding, upon reoxidation, of the reduced tyrosine modified-inhibitor is greatly diminished. Tyrosine modification can be prevented by carrying out the acetamidomethylation reaction in the presence of excess anisole. The stability constants and the standard free energies of binding of the complexes between trypsin and the native and the tyrosine modified-inhibitor have been determined.


Purification of a lysinonorleucine cross-linked peptide fraction from porcine aorta elastin. Highly purified elastin from porcine aorta was submitted to elastase digestion. The enzymic products were subjected to gel filtration on Sephadex G 25. The excluded fraction was then submitted to thermolysin digestion and gel filtration. The excluded fraction was submitted to partial acid hydrolysis and gel filtration. Several sub-fractions were obtained. The F3 subfraction containing cross-linking agents (desmosines and lysinonorleucine) was finally subjected to ion exchange chromatography. A highly enriched peptide fraction containing lysinomorleucine was obtained and then purified by preparative electrophoresis. The ratio of enrichment passed from 2 residues of lysinonorleucine (expressed as lysine) from starting material (elastin) to 178 out of 1,000 residues in the final step. In this peptide fraction, if we express in molar ratio and consider the amount of lysinonorleucine is one residue, the following amino-acid composition is Pro:3, Gly:1, Ala:2, LNL:1, Lys:2. No traces of desmosines are detected. The role of lysinonorleucine in bridging functions in elastin is discussed.


Circular dichroism of human pituitary luteinizing hormone and its glycopeptides. Curve resolution and band assignments to the peptide chromophore, aromatic residues, disulfides, and N-acetylated amino sugars. The circular dichroic (CS) spectrum of the glycoprotein hormone, human pituitary luteinizing hormone (hLH), has been determined between 195-320 nm and resolved into gaussian constituents. Below 230 nm the CD spectrum is characterized by a negative extremum at 207 nm with a shoulder at 217 nm. Resolution into gaussian constituents of the 200-230 nm CD spectrum resulted in two resolved negative bands, one at 206 nm and the other at 215 nm. The latter band is assigned to beta-structure which is estimated to be about 25%. The 206 nm resolved band is assigned to the N-acetylated carbohydrate groups (e.g. N-acetyl glucosamine, galactosamine, and neuraminic acid). This is based partly on the evidence that the CD spectrum of the hLH glycopeptide fraction (prepared by a pronase digestion of s-carboxymethylated hLH) exhibited a negative extremum at 207.5 nm, which is close to the resolved 206 nm band in hLH. Above 230 nm the CD spectrum is characterized by a negative extremum at about 275 nm. Most of the ellipticity in this region is attributed to the disulfides in hLH. Both strong acid (0.1 N hcl) and concentrated guanidine hydrochloride (4 M) affect the ellipticity in the vicinity of 275 nm, but only the latter (as well as concentrated urea) has a major effect on the CD spectrum below 230 nm indicating extensive conformational changes. There is, however, some loss of beta-structure in 0.1 N hcl. Thus, it appears that the conformation of the hLH subunits in these subunit-dissociating agents is rather different. There was no dramatic change in the magnitude of the 207 nm extremum of native hLH between 10-50C.


Solid phase synthesis and some pharmacological properties of 8-D-homoarginine-vasopressin and 1-deamino-8-D-homoarginine-vasopressin. Fully protected 8-D-lysine-vasopresin and 1-deamino-8-D-lysine-vasopressin were synthesized by the solid phase method. Selective removal of the lysine protection and reaction with 1-guanyl-3,5-dimethylpyrazole converted D-lysine into D-homoarginine. The title compounds were then obtained by treatment with sodium in liquid ammonia and oxidation in dilute aqueous solution. Although the antidiuretic activities are lower than for the corresponding D-argining derivatives, the even lower pressor effects make the new analogues highly specific antidiuretic agents. The A/P ratios for 8-D-homoarginine-vasopressin and its 1-deamino derivative are 100 and 3,300, respectively.


Preparation of chromophoric substrates for the glutamoyl specific staphylococcal protease. The synthesis of chromophoric substrates allowing an accurate determination of the staphylococcal protease activity is described. BOC-L-Glu-OPh, BOC-L-Phe-L-Glu-OPh, BOC-L-Ala-L-Glu-OPh, BOC-L-Ser-L-Glu-OPh and Z-L-Glu-pNA were prepared. Kinetic parameters of the staphyloccal protease-catalysed hydrolyses of these substrates are compared. In every case the dipeptide ester substrates lead to a lower catalytic efficiency (kcat/Km ratio), compared with either BOC-L-Glu-OPh or Z-L-Glu-OPh, mainly because of an increase in the Km value. Like other serine proteinases, the staphylococcal protease exhibits a high ratio of eeterase to peptidase activity, the kcat/Km ratio being 2.6 X 10(5)-fold higher with the Z-L-Glu-OPh than with the Z-L-Glu-pNA.


Studies on pituitary lactogenic hormone. The primary structure of the porcine hormone. The complete amino acid sequence of porcine lactogenic hormone has been elucidated. It consists of 199 amino acid residues with three disulfide bridges formed by residues 4-11, 58-174, and 191-199. The two tryptophan residues are in positions 91 and 150. A high degree of homology exists between the known structure of ovine prolactin and the porcine lactogenic hormone. This led to a re-examination of residues 82-90 in the ovine prolactin structure, where an extra leucine was found in position 88.


Studies on trypsin inhibitors. Part IV, Synthesis of the protected undecapeptide (sequence 25-35) of porcine pancreatic secretory trypsin inhibitor II (Kazal). Synthesis is described of the protected undecapeptide tert-butyloxycarbonylisoleucyl-threonyltyrosylserylasparaginyl-gamma-tert-butylglutamyl-S-acetamidomethylcysteinyl-valylleucyl-S-acetamidomethylcysteinylseriny hydrazide corresponding to positions 25-35 of the amino acid sequence of porcine pancreatic secretory trypsin inhibitor II (Kazal). The hepatapeptide free base methyl asparaginyl-gamma-tert-butylglutamyl-S-acetamidomethylcysteinylvalylleucyl-S-acetamidomethylcysteinylserinate (sequency 29-35) was acylated, by the azide procedure, with the tetrapeptide tert-butyloxycarbonl-isoleucylthreonyltyrosylserine hydrazide /sequence 25-28) and the resulting tert-butyloxycarbonylundecapeptide methyl ester was transformed into the corresponding hydrazide by hydrazinolysis. The stereochemical homogeneity of the final product was assessed, after partial deprotection with aqueous 90% trifuoroacetic acid, by digestion with papain and aminopeptidase M followed by quantitative amino acid analysis.


High tibial osteotomy for osteoarthritis of the knee: a changing concept. High tibial osteotomy is most successful in patients with osteoarthritis with mild varus deformity which is not associated with subluxation of flexion contracture. Success in the valgus knee is limited. If tibial osteotomy is indicated, undercorrection is undesirable in the varus knee while overcorrection is undesirable in the valgus knee.


Subcutaneous rupture of the Achilles tendon. A series of 27 ruptures of the tendo Achillis are reported. Samples of tendon tissue, taken both at the site of rupture and at a distance from it, were histologically examined. Four different surgical procedures were performed. All patients showed marked degenerative changes of the tendon tissue. No definite relationship was found between the surgical procedure employed and functional results obtained. These were rated as excellent in 12 cases, good in 11 and fair in four. Only minor complications attending the operation occurred. We emphasize the superiority of open repair over conservative treatment.


Fatal fat embolism associated with polymethyl methacrylate bone cement. Three fatalities after procedures involving the use of polymethyl methacrylate cement are reported. Autopsy studies showed varying degrees of pulmonary fat embolism although lack of correction of operative blood loss was the major factor.


Use of acrylic of low polymerization after heminandibulectomy. Two cases of mandibular reconstruction are presented in which a premolded prosthesis of acrylic of low polymerization covered by silicone rubber was used. We believe that the good results obtained were not only due to the material used, but also because the periosteum and meniscus were preserved in the joint space.


Temporomandibular ankylosis.

Temporomandibular ankylosis. 1. Intraarticular hydrocortisone and passive manipulation under general anesthesia does not help mobilize the jaw. 2. Condylectomy gives good results in selected cases, where the duration of ankylosis is minimal. 3. Longer duration requires wide excision of the condyloid as well as coronoid processes. The opposite side also must be operated on in most late cases. A gap of more than 2 cm is left in order to avoid recurrences. 4. Postoperative physiotherapy of the jaw in the form of active and resistive movements of the jaw, if started early, gives good functional results and helps recover the function of the chewing muscles. 5. Functional results are good even without interposition of soft tissues or use of prostheses. 6. Skeletal traction after operation is unnecessary. 7. The results of adequate resection of bone followed by early and adequate physiotherapy are very gratifying to the patient.


Congenital dysplasia of the hip in northern India. The incidence of congenital dysplasia of the hip in Northern India is 0.8% in infants born in two hospital maternity units which are interested in detecting this condition.


Fatigue fracture of the femoral neck.

Fatigue fracture of the femoral neck. Fatigue fractures in the necks of femurs removed from fresh cadavers were produced experimentally by applying repeated compression and rotational stresses. We found that a load applied once in a physiological direction may be within the ultimate strength of the bone, but may produce a fracture if applied repeatedly. With increased age, chances of fatigue failure increased age, chances of fatigue failure increased and loads of even physiological magnitude produced fatigue fractures. This may also happen during life. The subcapital fractures produced in these specimens were similar to those seen in patients in a clinical practice. This has been reported many times in new young military recruits. In individuals undergoing unusually strenuous exercises, the stress concentrations can be unusually high.


Intravenous infusion anesthesia for upper extremity surgery. Regional anesthesia by intravenous infusion was successfully performed in 126 cases with only three complications. The technic is particularly useful for outpatient surgery and for cases where general anesthesia is unsuitable. This type of anesthesia is not for routine use. Proper equipment and personnel must be available. We do not recommend its usage in very obese patients or where absolute hemostasis is essential. Caution is urged in cases where edema has developed as the absorbtion of local anesthetic is not predictable.


Unusual lesions of the patella.

Unusual lesions of the patella. We have reported 13 cases illustrating various rare lesions of the patells and discussed the incidence and treatment and reviewed other reported cases of each type of lesion.


Complementary retroperitoneal lavage.

Complementary retroperitoneal lavage. Surgery is required in acute necrotic pancreatitis when ominous signs appear after three days of intensive medical therapy. The surgical procedure is outlined and retroperitoneal lavage is proposed as a necessary complement. A protocol for this procedure is given. We believe that removal of necrotic tissue is not synonymous with partial pancreatectomy. Lavage completes what we consider "lesional pancreatic surgery" by diluting, buffering and washing out residual caustic tissues. The resulting fistula is directed and spontaneously dries up in most cases.


Unusual lesions of the clavicle.

Unusual lesions of the clavicle. Eight types of rare clavicular lesions comprising 41 cases with their treatment and results are presented. Total claviclectomy was performed in ten cases of pyogenic osteomyelitis with no resultant functional disability to the patient. We advocate this as the treatment of choice.


Lumbar sympathectomy in the treatment of uncomplicated intermittent claudication. The precise role lumbar sympathectomy plays in the treatment of uncomplicated intermittent claudication continues to be debated. Past experience at this institution indicates that lumbar sympathectomy has a definite positive role in uncomplicated intermittent claudication. A critical analysis of the 86 patients included in this study adds further support to this contention. Properly selected patients who undergo complete anatomic sympathetic denervation of the lower limbs continue to have very satisfactory results, as they have since 1947 in this institution.


Modified pancreatoduodenectomy for carcinoma of the head of the pancreas: preliminary report. A new technic for restoration of gastrointestinal continuity following pancreatoduodenectomy for cinoma of the head of the pancreas is described and the advantages discussed. We believe this method prevents any leakage of the anastomosis and reduces mortality and morbidity because of better drainage. Also, by avoiding stasis, infection, which is a common complication in this kind of surgery, is prevented.


Clinical and therapeutic applications of a new pathogenetic theory on the dumping syndrome. Based on experimental and clinical studies we have deduced the existence of a neuro-osmotic dumping syndrome and a truly osmotic dumping syndrome that differ in pathogenesis and clinical symptoms.


Intestinal parasitic infestation among parturients in Trinidad and Tobago. 1. In this limited survey, 42.5% of parturients were found to be infested with intestinal parasites. 2. Hookworm, Trichuris and Entamoeba infestations were the most common. 3. Race, social class and locality influenced in the incidence of the different types of infestations. 4. Those infested with hookworm had longer labor, smaller babies and higher incidence of prematurity.


Intussusception in adults.

Intussusception in adults. A review of intussusception in adults treated over a 12-year period is presented. In most cases intussusception was ileocolic in type and symptoms were nonspecific. Benign neoplasms and idiopathic causes were more prevalent in small bowel cases while malignant neoplasms were more common when the colon was involved. In 38% of the cases, no obvious cause for the intussusception was found. All cases were treated surgically, usually by reduction or resection or both.


Primary liposarcoma of the skull.

Primary liposarcoma of the skull. Only 14 cases of primary liposarcoma of bones have been reported in the available world literature. This is the first report of its origin from the calvaria. The tumor showed excellent response to deep x-ray therapy.


Pattern of intestinal obstruction in Khartoum. We operated on 138 patients for intestinal obstruction. The different causes are discussed and their pattern of occurrence is compared with that reported from England and other African countries. The 27.6% mortality rate was closely associated with the 28.2% incidence of gangrene. Factors which may lower this high mortality rate are enumerated.


Kidney stone matrix. Differences in acidic protein composition. Organic matrices from kidney stones of several mineral compositions (calcium oxalate, uric acid, and apatite-struvite) were isolated and found to contain an abundance of acidic amino acids. The calcium oxalate stones contained 50% aspartic and glutamic acid residues while the uric acid stones contained over 65%. The apatite-struvite stones had only 30% of these two amino acid residues and also contained 20% glycine residues. Since the specific amino acid composition differed for stones of different mineral content, it was felt that the organic matrix might play the role of a nucleating agent.