Development and clinical evaluation of novel radioimmunoassay for the detection of hepatitis B antigen.

Australia antigen or hepatitis B antigen (HBAg) is intimately associated with acute and chronic forms of serum hepatitis. It appears in the patient long before the onset of acute serum hepatitis. Transfusion of blood containing this antigen frequently leads to hepatitis in the recipient. Due to this, numerous researches have been done to develop a sensitive diagnostic test for detecting the presence of this antigen. To this date, radioimmunoassay has shown better sensitivity and specificity. The test developed at Curtis Laboratories is a human double antibody solid phase radioimmunoassay which is called RIAcT:HBAg. Human HBAb was absorbed on a polyethylene pellet and placed in a glass test tube to prevent radioactive antibody from binding to it. In this procedure, 0.2 ml of patient unknown serum or plasma is placed into the test tube containing the antibody-coated pellet. The vials with the unknown samples and proper controls are incubated in a water bath at 37 degrees C for 2 hours, washed and incubated further with iodinated human antibody for 90 minutes at 37 degrees C. At the end, the tubes are washed again and counted. The level of antigen present is determined by the radioactivity present. The counts of the patient samples that are at least twice the mean counts of the negative controls are considered positive for HBAg. The sensitivity of this test was checked many times by determining the reactivity of various samples from the Bureau of Biologics. The lowest threshold of this RIA procedure has been 1.07 X 10(10) particles per ml.


Change in prevalent subtypes of HBsAg in acute hepatitis B infections and its implications.

Subtyping of hepatitis B antigen in patients with acute hepatitis B revealed subtype ay in 75% while subtype ad was much more common in chronic asymptomatic carriers of the antigen (in 81% of blood donors). In contrast to blood donors, a change of the prevalent subtype was noted in acute hepatitis patients between 1970 and 1974. Before May 1972, subtype ad was found in 67% of patients, whereas later subtype ay dominated in 93%. An unequal distribution of subtypes between acute and chronic forms of hepatitis B infections has been explained by differences in the virulence of virus strains. Our results suggest that the higher prevalence of subtype ad was also common in acute infections. The change of the prevalent subtype in acute infections may be attributed to differences in contagion rather than differences in virulence of virus strains.


Distribution of HB antigen carriers in the population of Budapest aged 0 to 85 years.

The authors investigated serum samples of 5,000 inhabitants aged 0-85 years in a crowded industrial area, by means of the counterimmunoelectrophoretic method. They evaluate their results in relation to age, sex and occupation. They compare them with data of Hungarian and several other authors. Some possible reasons of the differences are discussed and the importance of non-parenteral transmission is emphasized.


Significance of the presence of the various subtypes for the preparation of HBAb.

The superficial Australia antigen is essentially bound to the small particles and possesses different antigenic specificities 'a', 'd', 'g', 'w', and 'r'. The population which we tested was characterized by the major frequency (more than 90%) of the 'ag' subtype with respect to the 'ad' subtype. The doses have been achieved using the solid phase RIA, modified according to Ling, in which the specificity of the antisera anti 'a', 'd' and 'g' was obtained by means of the absorption and dissociation of the antigen-antibody complexes. The investigations which we carried out to establish the avidity of the various anti-HBs anti-sera used for research into the Au antigen, have established that in general the avidity is maximal for the anti-ad and minimal or absent for the anti-ag. From this arises the necessity for the preparation of an anti-HBs anti-serum 'pool' consisting of healthy and hepatitic donors, with an optimal relationship between the subtypes 'ad' and 'ag'. The subsequent research carried out using the true subtypes in order to establish a relationship between the subtype and the presence of the anticore antibody on RIA revealed the lack of any correlation.


Hepatitis B core antigen and antibody.

Two distinct antigen-antibody systems are associated with the hepatitis B virus: the hepatitis B surface antigen (HBsAg, Australia antigen) and antibody (anti-HBs) and the recently characterized hepatitis B core antigen (HBcAg) and antibody (anti-HBc, anti-core). HBsAg is found in the serum of patients acutely or chronically infected with the hepatitis B virus (HBV) and is represented on electron microscopy of such serum as 20 nm spherical and tubular structures and as the surface component of the 42 nm Dane particle. The core antigen of the HBV is associated with a 27 nm viral nucleocapsid particle present in the nuclei of infected hepatocytes and with the inner, core component of the Dane particle. These two HBV antigens have been shown to be antigenically distinct by immune electron microscopy. Immunization studies have demonstrated that both antigens are capable of eliciting distinct, humoral and cell-mediated immune responses in guinea pigs. Analysis of immune responses to HBcAg during the course of type B hepatitis provides a clearer understanding of this disease and anti-HBc may serve as a sensitive marker for the presence of the chronic HBsAg carrier state.


Purification and characterization of the 'core' particles and their potential diagnostic use.

The basis of the postulated correlation between 'core' particles and 'Dane' particles, of their different localization in the hepatocyte and of the different relationships is the mechanism of replication of the infecting material, interesting diagnostic possibilities appear evident in the hepatitis B field. Anti-core antibodies would be alone present in the serum of the hepatitic patient, in the acute or convalescent stage, whilst in the serum of the polytransfused patient they would appear in association with the antibodies directed against the superficial antigen of the Dane particles. The 'core' particles have consequently been concentrated and purified by means of 'coat' lysis using tension actives and density gradient ultracentrifugation. With such material and a hepatitic serum purified antibody, it has been possible to prepare a radioimmunoassay with which a 'screening' may be performed to evaluate the possible diagnostic use.


Comparison of DNA polymerase and radioimmune assays for the detection of hepatitis B antigens and antibodies.

In acute cases of hepatitis, DNA polymerase activity was found 2 to 3 times more frequently than positive radioimmunoassay. For each case the DNA polymerase reactivity was shown to be associated with hepatitis B antigens. Inhibitors to this DNA polymerase, with properties of IgM and IgG antibody, were found in 13 of 34 cases of acute hepatitis but only in 1 case out of 22 of cirrhosis. During the course of the acute disease these antibodies were detected 3 times more frequently than those to HBs antigen; the two types of antibodies were almost always found separately in different patients, those to DNA polymerase were apparently transient and developed earlier since they were found as early as 3 days after the clinical onset and no later than the 6th week following the onset.


Core antibodies and other parameters of hepatitis B virus infection in a tropical population.

HBsAg (80% of ay subtype) was detected with radioimmunoassay in 10% of a sample of the healthy population in Mopti, Mali, and in 6% with counterelectrophoresis. Anti-HBs was detected in approximately 57% of adults, and anti-HBc in 50%. Anti-HBs was not always detectable in anti-HBc positive sera with the methods used, and vice versa. This accounted for up to 85% of an adult group showing anti-HBs, anti-HBc, or both. While anti-HBc percentages climbed more gradually with age, and 'adult' percentage of anti-HBs was reached at about 3 years of age, and there were no significant differences in the age distribution of HBsAg.


[Culture of hepatitis virus B].

For the last 25 years, numerous attempts have been made to isolate the HBV agent responsible for hepatitis B by means of cultures 'in vitro'. We have undertaken longterm cultures of children's hepatic tissue (C.H.), conjunctive tissue (human adult H.A.F. and human embryonic fibroblasts H.E.F.) and KB cells; these were put in the presence of 7 sera HB + rich in Dane particles. These cells were trypsinized twice a week for almost 3 months and did not present any cytopathogenic effects. Electromicroscopy revealed, 15 days after infection, the presence of icosahedral particles (25 to 27 nm in diameter), free or in dense clusters, but more often empty (20 nm in diameter). These structures seemed to be made up of an assembly of capsomers approximately 5 nm in diameter, joined together in fours to form a ridge. Older cultures revealed clusters of icosahedrons some of which degenerated spontaneously; others were surrounded by proteinic structure having a fringed aspect. Certain rare particles of 35 to 45 nm in diameter are similar to full Dane particles. EID immunological results were positive in the case of sera of patients convalescent from hepatitis B, containing anti-HBc antibodies, on C.H. cells the 27th and 40th days, and negative with anti-HBs antibodies. By immunofluorescence we observed 12 to 20 days after infection of the cells, a clear fluorescence at first nuclear, then essentially cytoplasmic, by means of fluorescent anti-HBc sera of human or animal origin. With the fluorescent anti-HBs antibodies, the reaction is weak and solely cytoplasmic although in DRI, with H.E.F. and KB cells, we obtained from time to time weakly positive results in HBs. The relations between the morphological structures and the immunological results observed are discussed.


DNA and DNA polymerase of a virus-like particle in hepatitis B.

One of the particulate forms bearing hepatitis B antigen in human blood, the 42 nm Dane particle, has been shown to contain a small circular-stranded DNA with a molecular weight round 1.6 X 10(6) daltons and a DNA polymerase. The circular DNA functions as a primer-template for the DNA polymerase. The circular DNA, the DNA polymerase and the DNA product of the enzyme reaction appear to be internal components of the 28 nm core of the Dane particle. Dane particle cores labeled with H3 in a DNA polymerase reaction have been used in a sensitive double antibody precipitation assay for the antibody against core.


Hepatitis B core antigen in the immunosuppressed chimpanzee.

Two chimpanzees with low levels of anti-HBs developed increased antibody titres but showed no antigenemia after i.v. administration of 10 ml infective chimpanzee serum. Treatment of a chimpanzee (also possessing anti-HBs) i.m. with Cyclophosphamide plus Prednisolone for 3 weeks starting 2 days before the challenge with infective serum resulted in detectable circulating HBsAg by day 67. By day 95, the HBsAg concentration had increased to 17 times a human Ag reference plasma and low titres of anti-HBc were also present, but liver enzyme levels did not become abnormal until day 107. Since the circulating HBsAg concentration decreased gradually to 2 times the reference plasma by day 168, the animal was again immunosuppressed using oral Prednisolone alone for 7 weeks and then sacrificed. The liver yielded useful quantities of purified HBcAg while the plasma was an excellent source of Dane particles and HBsAg (41 X human reference plasma). This success could not be reproduced in 4 immunosuppressed cynomolgus monkeys. Our studies, therefore, demonstrate the value of the immunosuppressed high-order primate in the initiation and modulation of hepatitis B.


Immunogenetic factors in thalassemia and hepatitis B infection. A multicentre study.

Geographical variations in the prevalence of hepatitis B infection were found in association with an increased opportunity of exposure to this virus in patients with homozygous beta-thalassemia in Cyprus, Greece, Sardinia and the United Kingdom. No association was found between the Gm phenotype of the individuals investigated and susceptibility to hepatitis B virus using HBsAg as a marker, or the ability to make antibody to it. Variation in the distribution of Gm haplotypes does not appear to be a feasible explanation for variation in anti-HBs and HBsAg frequencies in a series of highly transfused populations.


The variety of responses within a community to infection with Australia (hepatitis B) antigen.

There is a striking difference in the response of individuals to infection with HBV. Some individuals develop hepatitis, others form antibody which may persist for a long period, and others may become persistent carriers of Australia antigen. These responses may give some indication of the immune status of an individual and of populations relative to this agent. As such, it could provide a guide to the use of vaccines against hepatitis in areas of common occurrence of the disease. We have completed a survey of a community in northern Greece for the three factors. A very large percentage of the population have evidence of exposure and this varies with age and sex. Many responded by forming high titers of antibody but a considerable percentage developed hepatitis and/or became carriers. Some appeared to have complex. Most developed antibody against the prevalent antigenic type (ay) but in many cases antibody against the rare antigenic type (ad) appears to have been formed. Host characteristics appear to have influenced the nature of the response to the infection. The effects of vaccines on carriers is not known and conceivably could be detrimental (i.e. by formation of complexes). It would be important to learn more of the biology of Au in populations and individuals in connection with the widespread use of vaccines.


Humoral and cellular immunity in guinea pigs to polypeptides derived from HBsAg-containing particles.

Recent studies in this laboratory have shown that purified HBsAg contains at least 7-9 polypeptide subunits ranging in molecular weight (MW) from 18,000 to 120,000; at least three of these are associated with carbohydrates. In addition, two glycosphingolipids have been extracted which are structurally similar to the blood group antigenic substance. The glycoproteins and glycolipids are of particular interest since it has been reported that carbohydrate is associated with the antigenic activity of HBsAg. Antisera to these subunits have been produced in guinea pigs. These results stimulated further 'in vivo' investigations into the possible development of a subunit vaccine which would exclude genes of a viral or cellular nature and would not be infectious. Guinea pigs were immunized with HBsAg, subtypes adw and ayw, and with two polypeptides (40,000 MW and 24,000 MW) in complete Freund's adjuvant. Control animals were inoculated with normal human serum or with polyacrylamide gel. Animals were boostered on day 14 and tested on day 30 for the development of cell-mediated immunity (CMI) by the macrophage inhibition assay, and for humoral immunity by PHA and RIA. Migration of peritoneal exudate (PE) cells was inhibited 30-88% in guinea pigs immunized with the 40,000-MW polypeptide when challenged with purified HGsAg, subtype adw or ayw, or with the 40,000-MW preparation, but the results were inconclusive with the 24,000-MW subunit. Similar evidence of CMI was observed with PE cells derived from guinea pigs inoculated with purified subtype-specific HBsAg or with the 24,000-MW polypeptide. Humoral responses were predictable, and showed a reasonable degree of correlation with the CMI response. Control animals failed to respond to challenge with purified preparations of HBsAg or the 40,000-MW polypeptide, but did show CMI with their respective immunogens and - in the case of guinea pigs inoculated with normal human serum - with the 24,000-MW subunit, suggesting that the latter may contain an antigenic determinant related to a human serum protein.


A case of posthepatitic HBs-positive chronic hepatitis with transient monoclonal gammopathy.

Report of a case of HBsAg-positive chronic aggressive hepatitis associated with a transient monoclonal gammopathy.


Half life of transfused anti-HBs.

During a prospective study in post transfusion hepatitis 222 blood units were given to 55 patients who underwent open heart surgery. Eleven of these blood units contained anti-HBs titered higher than 1:128 by passive hemagglutination and were given to 10 patients previously negative for anti-HBs. Eight of these 10 patients showed anti-HBs in the first serum sample obtained usually one to three days after surgery and a continuous decline in titre was noted at later bleedings until antibodies were no longer detectable 2 to 20 weeks after transfusion. In four patients from whom regular blood samples were available the half life of passively acquired anti-HBs was determined to be between 23 and 28 days. In two patients with serum anti-HBs before blood transfusion no change in antibody titre was noted over a period of six months.


Epidemiological distribution and significance of anti-HBs.

HBsAg and anti-HBs neutralizing antibodies were quantitatively detected and typed by solid-phase RIA procedures in various random populations and in commercial lots of standard immunoglobulins. The prevalence of anti-HBs was considerably higher than prevalence of HBsAg, and the specificities of HBsAg and anti-HBs overlapped within the single groups of population. The results obtained provided evidence that hepatitis B virus infections are wide-spread, particularly in communities, and that the distribution of subtypes varies according to geographical patterns. The degree of cross neutralization between homotypic and heterotypic subtypes was not quantitatively constant, implying that the amounts of the single reacting determinants probably vary among HBsAg particles. It is also stressed that a number of sera containing both HBsAg and anti-HBs escape identification, since a fairly long interval can elapse during which the decreasing levels of HBsAg and the increasing levels of anti-HBs equalize in the blood.


Anti-HBAg immunoglobulin after inoculation injuries.

A Medical Research Council study was begun in England and Wales in September 1973 to determine the incidence of hepatitis B infection among persons who reported certain specified accidents with material containing hepatitis B antigen and who were given anti-hepatitis B immunoglobulin within 14 days. It was not considered justifiable to include a control group. Anti-HBAg immunoglobulin was separated by cold ethanol fractionation from plasma in which hepatitis B antibody was detected by counterimmunoelectrophoresis. The plasma pools were examined for the presence of HBAg bu immune electron-microscopy and radioimmunoassay. The aimmunoglobulin was prepared as a 5 g per 100 ml solution and the titre of anti-HBAg measured by CIEP, RPH, and RIA. The usual dose was 500 mg-intramuscularly. The study includes a year's follow-up of each participant. Serum samples usually taken at four weekly intervals for six months, then at nine months and one year are tested, by routine methods, for abnormalities of liver function and for HBAg and anti-HGAg. Specimens are stored for further tests at the end of the study. In the first year of study 110 persons were enrolled: of these, 100 participants continued to be followed up, about half of them for more than six months.


Specific immunotherapy proposed for hepatitis B virus infection.

Immune response to hepatitis B surface antigen (HBsAg), coinciding with removal of HBsAg from the circulation, has been shown to be a cell-mediated response assessed by leukocyte migration inhibition assay. Immune response to HBsAg is a T-cell dependent phenomenon in the nude mouse model. Immunological tolerance in man appears to stem from the absence of cellular and humoral immune response to HBsAg, causing a chronic carrier state which serves as an epidemiological reservior for the transmission of viral hepatitis type B. Specific immunotherapy in hepatitis may consist of adoptive transfer of immunity to HBsAg. It is proposed to be accomplished by administration of anti-HBs, transfer factor or 'immune-RNA'. The efficacy and safety of 'immune-RNA' administration to chronic carriers can be validated by leukocyte migration inhibition techniques in vitro and in HBsAg positive chimpanzees in vivo.


Viral hepatitis type B: propects for active immunization.

Studies conducted in 1970 and 1971 with heat-inactivated MS-2 serum revealed that this active immunizing procedure was associated with a protective effect, a more attenuated hepatitis B infection and a decreased hepatitis B carrier rate. More recent studies have revealed striking differences in the response of unimmunized and immunized persons following a parenteral exposure to the MS-2 strain of hepatitis B virus. Serial tests for the detection of hepatitis B surface antigen (HBsAg), DNA polymerase activity, serum transaminase (SGOT), antibody to HBsAg (anti-HBs) and antibody to hepatitis B core antigen (anti-HBc) revealed the following findings. In serosusceptible unimmunized persons HBsAg was detectable about 4 weeks after exposure, DNA polymerase activity at about 6 weeks, abnormal SGOT levels at about 8 weeks, anti-HBc at about 8-10 weeks, and anti-HBs usually after 20 weeks. In successfully immunized persons HBsAg, DNA polymerase activity, abnormal SGOT levels, and anti-HBc were not detectable, evidence of a booster response of pre-existing or non-detectable anti-HBs was observed one to two weeks after exposure. Studies by various investigators have revealed that anti-HBs is associated with protection and resistance to reinfection. In contrast, anti-HBc is not protective and does not correlate positively with either resistance to infection or recovery from infection. The availability of sophisticated biophysical and biochemical techniques has enabled several investigators to prepare candidate inactivated hepatitis B vaccines from purified preparations of HBsAg. The successful propagation of hapatitis B virus infection to susceptible chimpanzees has provided an excellent animal model for the evaluation of hepatitis B vaccines. At the present time various investigators are studying the immunogenic and protective effect of the vaccine in these animals. Prospects for the development of a vaccine for the prevention of viral hepatitis type B are encouraging. It is extraordinary that this objective will be achieved in spite of the failure to isolate the etiologic agent in tissue culture.


Some considerations regarding active immunization with HBsAg.

Krugman has demonstrated that injection of heated serum containing HBsAg conferred some protection against subsequent challenge with live hepatitis B virus (HBV). It is likely that improved protection will result from injections for larger quantities of HBsAg. This can be readily done with purified antigen. We have carried out preliminary studies in mice to investigate the inactivation kinetics of HBsAg antigenicity and immunogenicity. Rates of inactivation of the two parameters by heat have been found to be parallel. Chimpanzees will be required to determine adequacy of inactivation procedures, and optimal dose and immunization schedule, prior to the initiation of human trials. The use of newly imported chimpanzees for this purpose may be hazardous. Of 27 chimpanzees recently tested in an exporter's compound in Sierra Leone, ten were found to circulate HBsAg and five additional had detectable anti-HBs, suggesting a high risk of hepatitis B transmission within the compound. Thus newly imported animals may be incubating HB infection, and could give rise to false positive results in inactivation trials, unless quarantined for a four to six month period prior to use. We have therefore established a laboratory for chimpanzee trials in West Africa, and plan to utilize only animals captured by trapping and held in strict isolation from the time of capture. It is unlikely that immunization against HBV will ever provide absolute immunity. Three chimpanzees who received a massive transfusion of infective plasma one year after resisting challenge with aliquots of the same plasma inoculated intraperitoneally, all developed severe hepatitis, accompanied in two cases by HBs antigenemia, despite strong anamnestic anti-HBs responses in all three animals.


A new particulate antigen present in serum.

A new antigen has been encountered in the serum of 10 blood donors. It is distinct from Australia antigen, consists of spherical particles 21-23 nm in diameter and the corresponding antibody is widely distributed in the population.


Selection of tests for hepatitis B antigen.

Three commercially available tests - two hemagglutination and radioimmunoassay were used to test sera received in a routine clinical laboratory. Their overall performance in terms of reliability and sensitivity was very similar and the yield of positive results was increased by about 25%.


A latex agglutination test for hepatitis type B antigen.

A new latex agglutination test system (ANTIGEX-TM) for the detection of Hepatitis B Antigen (HBAg) is described. It can be used to detect HBAg in either serum or plasma with a sensitivity comparable to that of complement fixation and about four times greater than counterimmunoelectrophoresis. The sensitivity and specificity of the system is illustrated by the test results obtained on the F.D.A. Bureau of Biologics Sera Panel No. 2 and on sera and plasma specimens from normal blood donors in the U.K. and the USA. Further applications of the technique, in the detection of HBAg in finger-prick blood samples, and in the rapid determination of antigen subtype ('ad' or 'ay'), are also described.


Involvement of complement and RA factor in non-specific agglutination of the latex reagent for HAA.

Heat inactivation of fresh sera reduced significantly the proportion of false-positive reactions when the latex agglutination test for the detection of HAA was used in a series of 454 sera, possibly because heating destroys the C'lq fraction of complement. Non-specific agglutination also occurred if RA factor was present in sera tested, a fact that has been previously reported. This rapid and simple test may be very convenient as a screening procedure, but all positive sera should be tested again by means of a more specific method, as well as for RA factor. Improvement of the reagent may result in the elimination of false-negative results, which were no-observed in this study but have been reported by others.


Specific detection of hepatitis B surface antigen utilizing chimpanzee and guinea pig antibodies in a solid-phase radio-immunoassay.

The test involves a primary incubation of sample in polystyrene tubes coated with avid guinea pig anti-HBs serum followed, after washing, by a secondary incubation with immunopurified I125 chimpanzee anti-HBs. The system was found to be suitable for automation of washing and counting. The Hebria kit eradicated the non-specific positives obtained in those tests employing antibody from only one animal species and had third generation sensitivity (35 positives out of 50 samples) with the U.S. hepatitis B reference panel No. 2. Dilution analysis of both antigen subtypes showed that Hebria was as sensitive as Ausria II. By absorbing monospecific guinea pig antisera prior to coating, the kit showed greater than 10-fold discrimination between the ad and ay antigen subtypes while retaining high sensitivity. The kit could also quantitate and subtype anti-HBs in plasma samples.


Standardized detection of hepatitis B surface antigen: determination of its serum concentration in weight units per volume.

The concentration of HBsAg in serum was determined in arbitrary units by quantitative immunoelectrophoresis and in units of optical density by UV-photometry of purified antigen. The HBsAg was purified from serum by gelchromatography and subsequently by isopycnic centrifugation in cesium chloride. The ratio of the immunoelectrophoretically measured concentration to the units of optical density was found to be constant in six different samples with both subtypes - ad and ay - and varying concentration. The concentration of protein in three purified HBsAg samples was determined after acid hydrolysis by automatic aminoacid analysis. The specific extinction was E1 280 mg/ml = 4,5 +/- 0,3. Thus the arbitrary units of immunoelectrophoresis were converted to concentration of HBsAg specific protein, expressed as mug/ml. In 540 Ausria positive serum samples, mainly from the beginning of an acute hepatitis, 10-40 mug/ml was the most frequent range of concentration. About 4% of the sera contained more than 100 mug/ml, about 20% contained between 0,005 and 0,5 mug/ml and were positive only in RIA. 10 mug/ml correlated with titers in complement fixation of 1:32/64, in counterimmunoelectrophoresis of 1:8 and immuno-diffusion of 1:2. The use of standards with a defined concentration of HBsAg would allow a better control of the sensitivity in qualitative tests and of reproducibility in quantitation.


Direct solid-phase radioimmunoassay (dSP-RIA) for detection of antibody to hepatitis B surface (HBs) antigen.

Using direct solid-phase radioimmunoassay (dSP-RIA) which was shown to be specific and about 10000 times more sensitive than immunodiffusion and even 10 times more sensitive than the radioimmunoprecipitation test used by us before, anti-HBs was detected in 90% of hepatitis B convalescents who were followed up for a one year observation period and who had eliminated HBs antigen from their sera (60/68). In 13% of these patients (8/60 anti-HBs was only temporarily detectable. In 50% of all patients who developed antibody within a two to twelve months observation period (n = 151), anti-HBs appeared within three to four months after height of disease. In 16.5% of patients with HBs antigen negative hepatitis (32/193) anti-HBs was observed to develop during convalescence indicating a type B infection despite a negative finding for HBs antigen in the acute stage of disease. Using dSP-RIA anti-HBs was found in 5.5% of voluntary blood donors (n = 1036), in 20.2% of hospitalized patients without clinical hepatitis (n = 252), and in 30.6% of hospital staff (n = 304). Anti-HBs increases with age: in 14 year olds we found 0.5%, in 60 year olds, 14%. The fact that at least 70% of the anti-HBs positive normal persons and medical staff groups had no history of hepatitis illustrates the frequency and importance of subclinical hepatitis B infections.


An assessment of the Pictorial Test of Intelligence for use with young cerebral-palsied children.

In order to determine the concurrent and predictive validity of the Pictorial Test of Intelligence for use with cerebral-palsied children, this test was administered to 46 cerebral-palsied children aged between four and seven years, together with the Columbia Mental Maturity Scale and Peabody Picture Vocabulary Test. The results, compared with a classroom Achievement Rating Scale, showed the Pictorial Test of Intelligence to be superior to the other two tests as a predictor of classroom achievement of the children in this study.


Effect of amphetamines in hyperkinetic children: stimulant of sedative? A pilot study.

Dextro-amphetamine and laevo-amphetamine were given in single equal doses to six hyperkinetic children. The effects on motor activity and palmar skin admittance (a.c. conductance) were measured. Only the 1-isomer resulted in a significant decrease in motor activity. Both isomers resulted in a significant decrease  in skin admittance, the 1-isomer having the larger effect. These findings offer no support for the hypothesis that hyperkinetic children are under-aroused and benefit from stimulation. They indicate that the beneficial effect of amphetamines is attributable to a sedative action.


The hyperkinetic child: a behavioural, electrodermal and EEG investigation.

Ten hyperkinetic children were compared with a matched group of normal controls on the basis of behavioural measurements, electrodermal recordings and spectral analysis of the EEG. All the behavioural measures (total motor activity, speed of movement, and noisiness) differentiated the two groups in the predicted directions. The groups did not differ significantly in respect of the skin admittance (a.c. conductance) level or the EEG alpha rhythm propagation time. These findings provide no support for the hypothesis that hyperkinetic children are under-aroused. The best differentiation between the groups was provided by the EEG coherence function. The hyperkinetic group showed a significantly higher coherence within the hemisphere than the controls at frequencies up to 8Hz. It is speculated that they may reflect diminised cortical processing of information in the patients by comparison with the controls.


Treatment of minor motor seizures with clonazepam.

Fifty-two patients (age-range four months to 17 years) with drug-resistant convulsive disorders were treated for up to four years (average 15 months) with clonazepam, a benzodiazepine derivative. The types of seizures suffered by these patients were atypical petit mal, akinetic, massive infantile spasms, mixed minor motor seizures, myoclonic jerks, psychomotor, classic petit mal and grand mal. Evaluation of seizure control by clonazepam showed that the large majority of patients had improved--complete control of seizures was achieved in 27 per cent, and a greater than 50 per cent control was achieved in 61 per cent of the patients. Ten per cent of the patients showed no change and one patient worsened. Eight patients were successfully managed on clonazepam alone. Lethargy and ataxia were common side-effects but usually they were transient. No serious organic toxicity was noted.


A longitudinal psychophysiological study of three-year-old Mauritian children: preliminary report.

The rationale for using the high-risk-group research design in the search for the aetiology of schizophrenia is described. A longitudinal study is being made of three-year-old Mauritian children who underwent a variety of physiological and psychological tests. Groups of these children, selected on the basis of their responses to electrodermal stimulation, are now attending nursery schools established on the island as part of this study. A control group of children do not attend nursery school. It is intended to follow-up these children over the next two decades. Mention is made of the benefits of this research to the development of para-medical and educational services in a developing country.


The development of auditory figure-ground discrimination and ear asymmetry under nonaural stimulus presentation.

The development of auditory word discrimination ability in normal male children has been investigated. 56 school-children (14 from kindergarten and 14 each from second, fourth and sixth grades) were asked to discriminate 50 single-syllable words presented against four levels of background noise (0, +10, +20, and +30). Age, noise level, and ear were all significantly related to auditory word discrimination scores. The results of the study show that with increasing age there is increased ability to discriminate words in the presence of noise. However, although the older the child the greater his ability to discriminate, a far greater effect than age on discrimination is the signal-to-noise treatment level, a finding which suggests that knowledge of the treatment condition may be a better predictor of auditory discrimination scores than a child's age. An additional finding was a general right-ear superiority in the presence of noise, most consistently found under the 0 signal/noise condition for children in kindergarten and in second and fourth grades.


Cerebro-facio-thoracic dysplasia: report of three cases.

Three children, two girls and one boy, are described as suffering from a new syndrome, 'cerebro-facio-thoracic dysplasia'. The characteristic findings are mental retardation, characteristic facies, narrow forehead, bushy eyebrows with synophrys, hypertelorism, broad nose, wide philtrum, triangular-shaped mouth, short neck, marked maxillary hypoplasia, a low hairline (especially posteriorly in the midline on the neck), brachycephaly, calcified clinoid ligements, and multiple bony abnormalities in the upper thoracic vertebrae and sometimes in the cervical region, together with a variety of deformities of the upper ribs. It seems possible that the condition is hereditary and that it may be inherited as a result of an autosomal recessive trait.


The influences of propranolol on serum gastrin concentration and hydrochloric acid secretion in response to hypoglycemia in normal subjects.

In ten normal subjects the effect of propranolol on serum gastrin concentration and HC1 secretion during insulin hypoglycemia (0.2 U/kg) was studied. Under the influence of propranolol (50 mug/kg intravenously in 3 min) the gastrin response to insulin was abolished. The insulin-induced HC1 secretion was reduced by propranolol from 38 to 18 mEq/2h (p less than 0.01). These results are compatible with the hypothesis of beta-adrenergic stimulation on serum gastrin concentration and HC1 secretion during insulin hypoglycemia.


Epidemiology of Oesophageal cancer in France. Increasing mortality and persistent correlation with alcoholism.

Recent data concerning the mortality from oesophageal cancer by régions and départements in France are reported (around 5,000 deaths yearly). It was decided to investigate whether any time trend had become apparent and whether the correlation with alcoholism could still be observed. Numbers of deaths from oesophageal cancer for the years 1961-1963 and 1967-1968, and from cirrhosis and alcoholism for 1967 and 1968 were collected as well as the corresponding population from the census figures of 1962 and 1968. A cohort analysis using the data for 5 reference years from (1951 to 1971) is presented. Among European countries, France has the highest mortality rate from oesophageal cancer in males (13.65/100,000) and very high sex ratio (12.88). The rates are particularly high in western areas, especially Basse-Normandie. Over the 20-year period from 1951 to 1971, a progress of 32.7% was observed in males. The increase in mortality in males is greatest in the youngest age groups. In 1967-1968, there is a strong correlation between oesophageal cancer mortality and mortality from cirrhosis and alcoholism, the latter being more significant.


The colon: Absorptive, seccretory and metabolic functions.

The role which the human colon fulfils in digestion and metabolism remains largely undocumented. Its capacity to conserve water and electrolytes is well known although how this is controlled is uncertain. In the animal kingdom, calcium and magnesium absorption from the colon are improtant as are absorption and synthesis of vitamins. The abundant microflora of the human colon gives it unique properties. Dietary residue is metabolised forming short-chain fatty acids, hydrogen, carbon dioxide and methane; whilst 20% of urea synthesised in man is broken down in the colon to ammonia, which is reabsorbed, and carbonic acid. The microflora also degrades a wide variety of organic compounds including food additives, drugs, bile salts, and cholesterol which may be relevant to the development of colon cancer. Regional differences in colonic function also exist making interpretation of data from this relatively inaccessible organ more difficult.


The motor-stimulating effect of natural motilin on the lower esophageal sphincter fundus, antrum and duodenum in dogs.

Natural motilin was investigated for its effect on gastrointestinal motor activity in anesthetized dogs. In doses starting from 10 ng/kg as single injection and 100 ng/kg-h as intravenous infusion, motilin induced phasic pressure changes in the lower esophageal sphincter (LES) and in the lumen of the fundus, antrum, and duodenum. Tachyphylaxis was seen in 6 out of 10 dogs. Motilin had no effect on isolated muscle strip preparations of the LES, fundus, antrum, and duodenum in this species.


Interactions between vagus nerve stimulation and pentagastrin or secretin on the guinea pig gallbladder.

In guinea pigs, anaesthetised with urethane, stimulation of the peripheral cut end of the cervical vagus activates cholinergic excitatory and non-adrenergic inhibitory fibres to the gallbladder. Central vagal stimulation reflexly contracts the gallbladder by an efferent pathway in the vagus and also activates an adrenergic, non-vagal inhibitory pathway to the gallbladder. Injections of pentagastrin (12-33 mug kg-1 s.c.) or secretin (0.1-0.2 U kg-1 i.a.), doses which produce no effect on gallbladder tone when administered alone, significantly increase the contractions evoked by vagal stimulation.


Experimental lymphatic obstruction and fat absorption in the rabbit.

Using a previously published procedure of experimental abdominal lymphatic duct obstruction, we were unable to confirm increased faecal fat excretion after the operation. Nevertheless, the completeness of the obstruction and the huge dilatation of the lymph ducts of the villi have been verified. A spontaneous seasonal variation of faecal fat excretion has been observed in the rabbit with a maximum in October and November.


Synthesis rates of albumin and fibrinogen during and after acute hepatitis.

Synthesis rates of albumin and fibrinogen were determined in six patients with acute virus hepatitis and in nine control patients using the 14C carbonate method of McFarlane. Five patients were restudied several weeks after complete recovery. The following results were obtained. (1) The mean albumin and fibrinogen synthesis rates in the control group were 238.8 and 23.5 mg/kg/day, respectively. (2) In two patients with mild courses of hepatitis and in one patient studied during the early phase of a rapid and fatal course the albumin synthesis rate during the acute disease did not differ from controls whereas fibrinogen synthesis rate was slightly increased. (3) Three patients with severe hepatitis showed a definite decrease in albumin and fibrinogen synthesis rates, the lowest value being 76 mg/kg/day for albumin and 6.3 mg/kg/day for fibrinogen. (4) The decrease in plasma protein synthesis correlated neither with serum transaminase or bilirubin levels nor with plasma albumin or fibrinogen concentrations during the acute phase. (5) The two patients with the lowest albumin and fibrinogen synthesis rates also showed a definite decrease of the prothrombin index during the acute phase. Both patients presented very prolonged courses of the disease. (6) In all patients studied twice, plasma protein synthesis rates were normal after recovery.


Biliary excretion of conjugated sulfobromophthalein (BSP) in constitutional conjugated hyperbilirubinemias.

Sulfobromophthalein (BSP) and its three major forms of conjugates were determined in bile or duodenal aspitate, plasma and urine following intravenous administration of free BSP and synthetic BSP-glutathione (BSP-GSH) in five patients with the Dubin-Johnson syndrome and two patients with the Rotor syndrome, using alumina column chromatography. It was found that in Dubin-Johnson patients the biliary excretion of conjugated BSP was selectively impaired, conjugated BSP increased in plasma replacing free BSP after 30 min, and plasma retention of BSP-GSH was greater than that of free BSP when administered intravenously. In contrast, biliary excretion of BSP and its conjugates was not impaired and regurgitation of conjugated BSP into plasma was minimal in the Rotor syndrome. Thus, these two constitutional hyperbilirubinemias can be separated by their basic defects in BSP metabolism.


Muscle cell electrolytes in ulcerative colitis and Crohn's disease.

The intracellular electrolyte concentrations in skeletal muscle from 15 patients with ulcerative colitis or Crohn's disease were examined. In patients with diarrhea not receiving potassium, muscle cell potassium and magnesium were low and sodium insignificantly elevated. By contrast, largely normal muscle cell electrolyte concentrations were found in patients receiving potassium or having a normal intestinal function. Supported by the existence of normal muscle cell electrolytes in a control group of patients receiving prednisone for extraintestinal diseases it was stated that glucocorticoid treatment does not affect the muscle cell electrolytes unless such treatment is effective in controlling the intestinal function. Potassium - and possibly magnesium - supplementations are recommended in cases with diarrhea.


Postmortem study of the prevalence of gallstones in Athens.

The prevalence of gallstones in Athens in 1973 was studied in an autopsy survey that covered 1,448 subjects of all ages and both sexes. Gallstones were found in 5.2% of all subjects and in 6.2% of subjects older than 40 years. The prevalence was higher in females and considerably higher in persons who died from cirrhosis of the liver or from ischaemic heart disease.


Viscosity and osmolality of abnormal biles.

The mean osmolality of bile from the common bile duct is similar to that of bile obtained from the gallbladder of patients with non-functioning gallbladders, and both means are significantly lower than the mean of bile in functioning gallbladders. The mean viscosity of bile from both functioning and non-functioning gallbladders is on average considerably greater than that from the common bile duct, and the mean viscosity of bile from non-functioning gallbladders is greater than that from functioning ones. The presence of much mucus in gallbladders containing stones is likely to account for these differences in viscosity. The composition of the gallstones does not appear to have any influence on these observations, but the number of patients in each group is too small for the differences to be significant.


Gastric secretion in rats after infusion of stimulants into the abdominal vena cava and aorta.

Pentagastrin or histamine were infused for 24 h by way of the abdominal vena cava or aorta into conscious rats bearing permanent gastric and vascular cannulas. The locus of infusion did not significantly affect the output and concentration of acid and pepsin, or the volume of gastric secretion.


Small and large vessel disease in the development of foot lesions in diabetics.

This paper describes studies undertaken to determine the role of vascular disease in the development of foot lesions in diabetics, particularly those with neuropathy. The ratios of ankle/arm and toe/ankle blood pressure were used as indices of large and small vessel disease respectively. Diabetic patients with lesions had evidence of more large and small vessel disease than patients without lesions. Patients with neuropathic lesions had evidence of more small vessel disease than patients without lesions.


Glucocorticoids and the fate of exogenous insulin in insulin-immunized guinea pigs.

Groups of normal rats and of normal and insulin-immunized guinea pigs were injected intravasculary with mixtures of radio-iodinated and unlabelled porcine insulins. No differences could be discerned between the fates of radio-iodinated and unlabelled insulins in normal or immune guinea pigs except with respect to the kidneys of normal animals, which seemed to accumlate TCA-precipitable radioactive substances devoid of immunoreactivity. The abnormally slow disappearance of insulin from the plasma and its persistent accumulation in the livers and spleens of immune guinea pigs were not affected by prior treatment for three days with large daily doses (ca 20 mg/kg body wt.) of hydrocortisone acetate.


The effects of glucose, N-acetylglucosamine, glyceraldehyde and other sugars on insulin release in vivo.

The specificity for carbohydrates of insulin secretory responses in vivo was studies. Test sugars were injected via a left femoral vein cannula into conscious rats. Blood samples collected over the ensuing 60 min via a left femoral arterial cannula were assayed for plasma insulin and glucose, and, in some experiments, for N-acetyl glucosamine. Whereas L-glucose or saline produced no significant changes in plasma insulin or glucose concentrations, D-glucose, N-acetylglucosamine, D-glucosamine, fructose, D-glyceraldehyde and DL-glyceraldehyde were potent secretagogues. Simultaneous injection of mannoheptulose abolished the insulinortopic action of glucose and N-acetylglucosamine, but not of DL-glyceraldehyde. Fructose, glucosamine, and DL-glyceraldehyde induced hyperglycaemia, but peak insulin concentraions occurred before any change in plasma glucose concentration. No evidence was obtained for a stimulatory effect of galactose on insulin release. Infusion for 60 min of N-acetyglucosamine produced a sustained elevated plasma insulin concentration and significant hypoglycaemia. The present in vivo results agree with previous in vitro observations and could indicate a role for sugars other than glucose in the regulation of insulin release.


Glucagon and insulin from lean rats and genetically obese fatty rats: studies by radioimmunoassay, radioreceptorassay and bioassay.

Insulin, proinsulin and glucagon extracted from lean rat pancreases were studied in radioimmunoassay, radioreceptorassay and bioassay systems. Extracted insulin behaved identically to a rat insulin used as a reference standard in radioimmunoassay. On the basis of its immunoreactivity, extracted insulin was slightly less potent (about 70%) than the rat standard insulin in competing with the binding of 125I-insulin to rat liver membranes (radioreceptorassay) and in stimulating glucose oxidation by rat fat cells (bioassay). Extracted glucagon and a pork glucagon used as a reference standard were indistinguishable in two radioimmunoassay systems for glucagon, in competing with the binding of 125I-glucagon to rat liver membranes (radioreceptorassay) and in stimulating adenylate cyclase in rat liver membranes (bioassay). Genetically obese rats (Zucker, "fatty") were compared to their lean littermates with respect to insulin, proinsulin and glucagon extracted from their pancreases. Proinsulin represented the same proportion of total immunoreactive insulin in both types of rats. In the radioimmunoassays, the radioreceptorassays and the bioassays, insulin, proinsulin and glucagon from obese rats were indistinguishable from insulin, proinsulin and glucagon from lean rats. It is concluded that the pancreatic hormones of obese ("fatty") rats possess the same immunoreactivity and biological potency as those of nonobese rats. This excludes the possibility that some alteration in the biological properties of pancreas insulin and/or glucagon of fatty rats could explain the metabolic abnormalities observed in this type of obesity.


Plasma glucagon levels in normal women during pregnancy.

Increased plasma pancreatic glucagon concentrations have been reported during various states of decreased glucose tolerance. In vitro studies have demonstrated that human somatomammotropin stimulates glucagon release. The present investigation aimed at evaluating the role of plasma flucagon in the insulin resistance associated with normal pregnancy. Postprandial samples of plasma were obtained from 156 pregnant women between the 5th and the 40th week of pregnancy and were assayed for blood glucose, plasma insulin, glucagon and free fatty acids. Plasma insulin showed a gradual increase during pregnancy, and reached its maximal values during the last trimester. A moderate but significant increase in plasma glucagon was present between the 16th and the 28th week of gestation, whereas during the first and the last trimester of pregnancy its concentration was similar to that in non pregnant women. Intravenous glucose tolerance was performed during the last trimester and in a group of non pregnant control women. The slight decrease in glucose tolerance and the marked hyperinsulinemia associated with late pregnancy were accompanied by a more rapid and more pronounced decrease in plasma glucagon. A rapid and sustained decrease in glucagon was also observed when plasma FFA were raised by the intravenous administration of a triglyceride emulsion and heparin. These data suggest that glucagon is not involved in the insulin resistance associated with normal human pregnancy.


The contribution of endogenous insulin secretion to the ketogenic response to glucagon in man.

The magnitude and direction of the lipolytic and ketogenic responses following exogenous glucagon administration is controversial and consideration of the possible role of endogenous insulin secretion upon these events has not been clarified. The present study examines the role of endogenous insulin secretion in modulating the net lipolytic and ketogenic activity of glucagon. Three groups characterized by different levels of endogenous insulin secretory capacity were studied. In all three groups, the responses in plasma insulin, betahydroxybutyrate, and free fatty acids were observed following bolus administration of 1.0 mug/kg glucagon. In the obese subjects with increased endogenous insulin secretion, glucagon administration resulted in a decline below basal levels of both free fatty acid and betahydroxybutyrate. In the diabetic subjects with no demonstrable endogenous insulin secretion, glucagon administration was followed by a rise in plasma free fatty acids and an exaggerated rise in plasma betahydroxybutyrate. The normal control group exhibited a response in betahydroxybutyrate midway between the obese and diabetic groups. These obwervations support the thesis that the magnitude of endogenous insulin secretion modulates the lipolytic and ketogenic actions of glucagon in man.


Metabolic studies in familial partial lipodystrophy of the lower trunk and extremities.

A familial syndrome of partial lipodystrophy inherited as a dominant trait is reported. Subcutaneous fat loss was confined to the extremities and trunk. Diabetes mellitus, hyperlipidemia, hepatomegaly and renal disease were very prevalent in this family. Metabolic studies were performed on 3 members. In vivo tests suggested that the remaining fat tissue responded normally to stimulators and inhibitors of lipolysis. In vitro incubation of the dystrophic fat tissue of one patient suggested that the intracellular pathways of lipid and glucose metabolism were normal. The pattern of subcutaneous loss of adipose tissue observed in this family may be due to sympathetic nervous system overactivity of certain non-contiguous dermatomes.


Variation in polyol levels in cerebrospinal fluid and serum in diabetic patients.

Cerebrospinal fluid (CSF) or CSF and plasma levels of sorbitol, 1,5-anhydroglucitol and myoinositol of diabetic and non-diabetic patients with normal kidney function and of diabetic and non-diabetic patients with impaired kidney function were measured by gas-liquid chromatography. The CSF sorbitol level correlated with the plasma glucose level (p less than or equal to 0.05) in diabetic patients with normal kidney function, having received insulin for less than 12 months. The correlation between CSF sorbitol and plasma glucose levels in patients not dependent on insulin was not significant. Sorbitol was not detected in the plasma. The highest sorbitol levels in CSF were seen in insulin-dependent diabetic patients with impaired kidney function. No rise was seen in non-diabetic uremia. 1,5-anhydroglucitol, normally present in plasma, was absent from CSF and plasma in diabetic patients receiving insulin. In non-diabetic uremic patients, 1,5-anhydroglucitol levels in CSF and plasma were lower than in healthy subjects, but htere was no correlation with plasma glucose levels. The myoinositol level was higher in CSF than in the plasma of both non-diabetic and diabetic patients with normal kidney function. Both plasma and CSF levels were significantly (p less than 0.001) elevated in diabetic as well as in non-diabetic uremic patients, the plasma myoinositol increasing relatively more than the CSF levels. The elevation of plasma myoinositol correlated with the elevation of plasma creatinine and thus also with the impairment of kidney function. Plasma and CSF myoinositol levels were not influenced by the plasma glucose level.


Stability of the 4th International Standard for Insulin.

The stability of the (W.H.O.) 4th International Standard for Insulin, has been assessed by accelerated thermal degradation studies. This is a crystalline preparation of insulin, freed from proteolytic enzymes, sealed in ampoules containing air and with a moisture content of 5--6%. Of the original biological activity 95.8 (92.8--98.9;P = 0.95)% was retained after storage for 12 years in the dark at 20 degrees C and 65.7 (63.4--68.1;P = 0.95)% after 14 years at 37 degrees C. Degradation rate constants were calculated from these data for the Standard at 20 degrees C and 37 degrees C, assuming first order kinetics. The degradation constant at 37 degrees C did not differ significantly from those found in earlier degradation studies at 37 degrees C over shorter periods, thereby supporting the assumption that the degradation of crystalline insulin, at least at 37 degrees C, is a first order reaction. Extrapolation of these data suggest that the Standard stored at -20 degrees C for 20 years would have retained at least 99.93% (P = 0.95) of its original activity and so for practical purposes can be considered to be stable.


Transfilter studies on neural induction in the newt.

In order to study the transmission mechanism of neuralising signals during primary embryonic induction, the interacting components (competent newt gastrula ectoderm and dorsal lip tissues) were separated by filter membranes of varying pore size. Nuclepore filters with nominal pore size from 0.1 to 8 mum were employed and the neuralising effect was shown to traverse all of these membranes. Electron microscopic examination did not reveal any cytoplasmic processes in the pores and the authors conclude that the morphogenetic signals are carried by transmissable compounds rather than through direct cytoplasmic contacts.


Transfer and localisation of maternal serum antigens by mouse preimplantation embryos and oviductal epithelium.

Two hours after systemic injection of bovine plasma albumin (BPA) into pregnant mice, albumin-like antigen was detected by indirect immunohistological methods within the cytoplasm of oviductal and preimplantation uterine embryos whether ovulation was spontaneous or induced by hormone injection. Although fluorescence, localising antigen similar to or identical with the systemically injected foreign protein, was present in embryos in all oviductal regions and at all cleavage stages, the intraembryonic location of the transferred serum molecules differed from embryo stage. Most ootids and two-celled blastomeres contained large intracytoplasmic areas of intense fluorescence randomly associated with non-fluorescent or dimly fluorescent areas in the same cell. By four- and eight-celled stages, albumin-like antigen was localised at the periphery of blastomeres; less was found deep within embryos. By morula and blastocyst stages, blastomeres differed from each other in fluorescence intensity although intracellular fluorescence was homogeneous. Transferred BPA antigen, present in both pronuclei, probably was absent from blastomere nuclei. Ootid zonae pellucidae contained BPA antigen but none was detected in zonae surrounding cleaving embryos. Little foreign albumin was detected in oviductal epithelium. It is concluded that morphological, as well as biochemical, differentiation occurs during mammalian cleavage and it is suggested that maternal macromolecular contributions to mammalian preimplantation embryos may be necessary for normal development in vivo.


[Clinical diagnosis of corrected transposition of the great vessels by bulbo-ventricular inversion].

29 cases of corrected transposition of great vessels (CTGV), of which 3 cases were isolated and 26 cases were associated with various malformations were studied. The aim was to make the diagnosis of CTGV were made evident, to differentiate those which came from the associated malformations. The specific signs for CTGV are as follows: 1) a slight systolic ejection murmur at the 2nd or 3rd left intercostal space along the sternal margin; 2) a loud 2nd tone in the same focal zone. Specific radiographic signs are that: 1) the aortic arch comes up to sternal-costal articulation; 2) the inferior left margin has a convex form; 3) in the right oblique-anterior projection it is possible to see 2 imprints on the opaque esophagus, which is represented by the aorta above and pulmonary artery below...


[Frequency of serologic conversion of complement fixing antibodies FC by respiratory viruses in cases of acute juvenile respiratory disease].

The complement fixation test, run on double samples of serum pertaining to 447 children hospitalized between January 1971 and June 1974 at Ospedale Maggiore in Novara, or checked in consulting rooms for acute phlogosis of the respiratory organs, has permitted to detect in 241 children, equal to the 53,9% of the total cases, a significant increase of antibodies for respiratory viruses (Influenza A, B, C, Parainfluenza 1, 2, 3. Respiratory syncytial virus, Adenovirus, Reovirus). Specifically, in the 40,0% of the total cases, the serological conversion was evidenced for one virus only, and in the 13,8% for 2-3 viruses, raising in this last cases, questions concerning the diagnosis at a serological level.


[The congenital coronary artery fistula. Observations on seven cases (author's transl)].

The authors report seven cases of congenital coronary artery fistula observed in the Cardiology Department of Padua Medical School. The cardiopathy, the anatomo-histological characteristics, evolution and embriology are described. The anamnesic, clinical, electrocardiographic, X-Ray, phonocardiogram, haemodynamic, and contrastographic findings for each case are given. They are discussed and interpreted in the light of the pathophysiology of the disease.


Aneurysm of the pars membranacea of the interventricular septum.

Six cases of aneurysmal formation of the pars membranacea septi (AMS) are presented. Three cases associated with a ventricular septal defect (VSD) and one with VSD, prolapse of the right aortic cusp and aorto-right ventricular fistula, were operated on and the aneurysm, unsuspected preoperatively, was found at the operating table. Two cases, both with aortic coarctation (CA), have been diagnosed on the basis of selective angiocardiography and operated solely for resection of the coarctation. In three patients a mild pulmonary infundibular pressure gradient was present at catheterization. This anomaly is considered rare. It has been reported to complicate surgery, may not require surgery, and is considered by some to be a prelude to spontaneous closure of a VSD. The English medical literature concerning aneurysm of the membranous septum has been reviewed and our opinions on the management of these patients are presented. Including our own six patients, we are aware of approximately one hundred and sixty five reported cases, with possibly an additional 16 cases to be added.


[Plasma concentrations of complement components C3, C4, and C3PA in juvenile nephropathies].

Complement profiles in children suffering for nephropathy have been investigated. The approach has proved useful in differentiating distinct nosologic entities. Plasma C3 and, to a lesser extent, C4 levels were found to be markedly reduced in glomerulonephritis and significantly increased in nephrotic syndrome. Although in both conditions plasma C3PA concentration ranges normally, additional data are request before assuming that alternate pathway is not involved. The extensive serial study of complement profiles should be widely adopted by clinicians managing nephropatic patients.


The "inverted" pattern of the normal ventricular pressure curves: a syndrome with two possible angiocardiographic entities.

The "inverted" pattern of the normal ventricular pressure curves (VPCs) -- as herewith defined -- is considered a characteristic finding in patients with uncomplicated transposition of the great arteries (TGA). In more than 2,000 consecutive cases of congenital heart disease, this pattern was recorded in 6 patients. In all of them situs solitus was present with a large atrial septal defect (ASD) as the sole and obligatory defect. It is shown by angiocardiography in the "systemic" (venous) ventricle in all patients, that this ventricle is: I) the angiographic right ventricle in 5 patients, and II) the angiographic left ventricle in 1 patient. In I TGA with atrio-ventricular concordance (classic TGA) is diagnosed, while in II atrio-ventricular discordance with no TGA (isolated ventricular inversion) is diagnosed. In both instances a Mustard-type of operation is indicated. "Inverted" patterns of different magnitude are found. Therefore, as long as the pattern of VPCs is "inverted", differential diagnosis should be made during life between I classic TGA, and II isolated ventricular inversion.


[Programmed atrial stimulation in the study of sinoatrial conduction in normal subjects (author's transl)].

Sinoatrial conduction was investigated in 18 normal subjects, using premature atrial depolarizations. The results obtained in this investigation were evaluated plotting the test cycle (expressed as difference between the basic sinus cycle and the test cycle as a percentage of the basic sinus cycle) as a function of the return cycle (expressed as difference between the return cycle and the basic cycle as a percentage of the basic sinus cycle). In normal subjects, premature atrial depolarizations elicited in the last 10-20% of the spontaneous sinus cycle, produced a progressive prolongation of the return cycle and the points correlating the return cycle index to the test cycle index fell above the diagonal of the plotting system. After earlier premature atrial stimulations, the return cycle stayed the same length, and the points correlating the return cycle index to the test cycle index fell along a line parallel to "y" axis (plateau). The mean value of the returning cycle (expressed as above) corresponding to the test cycles (evaluated as above) included in the first 5% of the "plateau" can be defined as the "sinoatrial conduction index". This index, the sum of conduction into and out of the sinus node, ranged from 79 msec to 187 msec. By assuming similar anterograde and retrograde conduction, the sinoatrial conduction time ranged from 39.5 msec to 97.5 msec (mean value 70 msec).


[High-frequency electric stimulation of the atrium in treatment of supraventricular tachycardias and atrial flutter].

The authors report the results of rapid atrial stimulation in the treatment of 12 cases of junctional tachycardia (J.T.), 20 cases of atrial tachycardia (A.T.) and 43 cases of atrial flutter (A.F.). Sinus rhythm was restored in 91.6%, 70% and 60.4% of the cases for J.T., A.T. and A.F. respectively, either when pacing was discontinued or following a period of atrial fibrillation (from a few minutes to several hours). Fifteen per cent of the cases of A.T. and 25.5% of the cases of A.F. turned into stable atrial fibrillation with reduction of ventricular rate. The authors believe that this technique is a valuable alternative to D.C. countershock when medical treatment has proved ineffective and when countershock may be hazardous.


[Correlation between chemical carcinogenesis and immune response].

Methylcholanthrene, in amount sufficient to induce tumors in 100% of treated animals failed to influence primary and secondary phases of antibody synthesis in mice immunized with sheep erythrocytes and human serum albumin. The early immune response in tumor bearing mice was also indistinguischable from that of normal animals, despite the presence of marked splenomegaly in the former group.


[The isoproterenol test in the electrocardiographic diagnosis of coronary insufficiency. Experience in 100 cases of ischemic cardiopathy].

Intravenous infusion of 10 to 30 gamma/min. of Isuprel for 3 to 7 minutes is accurate in diagnosing coronary disease in at least 80% of cases in our series of 100 patients with segmental coronary artery stenoses of 50% or more, demonstrated by coronary angiography. By comparing these 100 patients with a control group of 30 healthy subjects we can state that the late "ST" segment changes (persisting or appearing 3 minutes after stopping the infusion) are typical of coronary insufficiency. In patients without previous infarction, coronary insufficiency is expressed by a horizontal "ST" depression of 1 mm or more. In patients with previous infarction we observed either an "ST" depression or an "ST" elevation. The "ST" elevation, never observed in the control group, seems to have a different significance depending on whether or not a previous myocardial infarction has occurred. If there was no previous necrosis, severe coronary artery disease seems to be suggested and is a bad prognostic sign. This is not so if the patient has previously presented a myocardial infarction.


[The contractile function of the left ventricle in pure mitral stenosis. Echocardiographic analysis].

The left ventricular performance in patients with mitral stenosis has been assessed by echocardiography, through the measurement of various parameters, such as the stroke volume, the ejection fraction (EF), the mean circumferential fiber shortening rate (VCF), the cardiac work and the left ventricular mass. Among these parameters only EF and VCF are statistically lowered in mitral disease, in comparison with their value for normal hearts: the mean value of EF is 0.62 in healthy people and 0.49 in mitralic patients; the mean value of the VCF rate is 1.05 circ/s and 0.78 circ/s respectively. The cardiac work and power in the considered patients are on the average lower than normal values by about 10%, but not significantly so. A good correlation has been obtained between the left ventricular work and mass, whereas no correlation has been found between the ventricular wall thickness and the diameter of the left cavity, both measured at end-diastole. The quotient of the ventricular thickness and intracavity radius, in diastole, is equal to 0.38 in healthy people and 0.30 in mitralic patients. Lastly, the echocardiographic results and those of angiocardiographic literature were in agreement.


Coronary circulation in patients with and without coronary artery disease and the effects of chewable isosorbide dinitrate.

Coronary arteriograms of 38 patients with suspected coronary artery disease (CAD) were first evaluated to decide whether or not the disease was present and, if so, whether easily recognizable collateral channels were demonstrated. In this evaluation, we found CAD in 26 of the 38 patients. Eighteen of 21 patients with severe obstructions or occlusions had functioning collateral vessels. The films were then evaluated a second and third time to determine the effects of chewable isosorbide dinitrate (ISDN) on the coronary circulation. Visual inspection of the arteriograms revealed significant increases in 1) the apparent number and diameter of collateral vessels, 2) the opacification of vessels distal to occlusions, and 3) the diameter of coronary arteries following the administration of 2.5, 5, or 10 mg chewable ISDN. Computer analysis of the arteriograms showed an average 16% increase in the diameters of specific segments of major coronary arteries following ISDN. All patients showed some degree of vasodilatation following ISDN; however, patients without CAD consistently showed more vasodilatation than patients with disease. Mean aortic blood pressure decreased an average of 10% following ISDN. These results demonstrate that the chewable form of ISDN reliably dilates the coronary arteries in patients both with and without CAD and enhances the collateral circulation to the ischemic areas in patients with CAD.


[Study of the carotid pulse with lead II (acceleration carotidogram) in cardiovascular diseases. Introductory note].

The relationship between heart dynamic and the carotid pulse has been studied by using the second derivative as a function of the time of the carotid pulse. This method permits a detailed analysis of the systolic phase of the carotid pulse: in protosystole a positive wave preceeds a negative wave and in telesystole a negative wave preceeds a positive wave. The morphology of the acceleration carotid pulse (CDA Carotidogramma Di Accelerazione), represented by a series of formulas which relate between themselves the protosystolic and the telesystolic waves, is a characteristic as well as statistically significant in each class of the cardiovascular disease in which we examined aortic insufficiency and stenosis, mitral insufficiency and stenosis, aortic angiosclerosis and hypokinetic myocardiopathy. We propose the use of the CDA for a precise analysis of the process of contraction and relaxation of the left ventricle. Since the acceleration with which a phenomenon evolves is the expression of the strength that has generated it, the CDA is particularly effective for the study myocardial dynamic.


[The ventilation-perfusion ratio, with special reference to muscular exercise].

The values of VA/Q obtained at rest in 12 normal subjects undergoing cardiac catheterization in a supine position were between 0.63 and 1.695, with a mean of 1.142 +/- 0.295. The VA/Q values obtained in another 10 healthy subjects tested in a sitting position with a rebreathing method for calculating Q. were somewhat less scattered (between 0.77 and 1.50), and also lower (mean 0.975 +/- 0.210). A highly significant correlation was demonstrated during muscular exercise on the bicycle ergometer (sitting position), both between oxygen consumption and alveolar ventilation and between oxygen consumption and cardiac output. However, since at various submaximal work loads cardiac output increased much less than alveolar ventilation, the overall VA/Q ratio showed a progressive increment with increasing oxygen consumption.


[ABO blood-group phenotypes and pathogenesis of cardiovascular diseases. Congenital, rheumatic and coronaric heart disease and arterial hypertension (author's transl)].

Many cases of cardiovascular diseases have been examined in reference to the distribution of ABO blood-groups, in order to calculate the relative risk of disease and the hemogroupal distributive significance in our samples as related to those of other authors, using combined calculation. The analysis concerned the following cases: 746 with arterial hypertension, 3258 with congenital heart disease, 4503 with articular rheumatism, 1047 with acquired valvulopathia, and respective controls. It was found that blood-group phenotypes represent an important biophysiopathological action in regard to articular rheumatism and its cardiac consequences, in myocardial infarction and in hypertension, males only. On the contrary, no action in regard to congenital heart disease was found, with the exception of some single anomalies which have yet to be confirmed. This hemogroupal action greatly exceeds the one limited to the immunitary analogy and is a noticeable part of family heredity. It shows itself in: -- a significant negative association with group O and positive association with group A in the myocardial infarction; -- a significant negative association with group O and positive for the others in the valvulopathic (rheumatic) diseases; -- a positive association with A phenotype and negative with B in arterial hypertension, males only; -- no association with ABO blood-groups and congenital heart disease.


[Evidence for circadian variability in the renin-angiotensin II system (author's transl)].

Circadian variations of P.R.A. and angiotensin II were studied in three normal subjects and nine hypertensive patients. All data were analysed by a mathematical-statistical method called "cosinor". Analysis of results has shown: 1) Circadian rhythm of P.R.A. which in the normal subjects reaches its acrophase (maximum daily level) in the morning, while in the hypertensive patients the acrophase recurs in the afternoon; 2) Circadian rhythm of angiotensin II in the normal subjects, which is not detected in the hypertensive patients. Angiotensin I does not exhibit rhythm in both groups.


[Completely interrupted aortic arch, ventricular septal defect and patent ductus arteriosus. A complete surgical correction in one procedure (author's transl)].

The authors present a case of completely interrupted aortic arch (Type B) associated with ventricular septal defect and patent ductus arteriosus in a child 3 years and 10 months old, who was successfully subjected to complete surgical correction in one procedure. The pre- and post-operative electrocardiographic and hemodynamic data are evaluated. The authors draw attention to the extreme rarity of surgical resolution, in one procedure, of this type of congenital heart disease.


[Myocellular necrosis by cathecolamines in pheochromocytoma (author's transl)].

A case of myocellular necrosis by cathecholamines in a patient presenting pheocromocytoma is reported. Death was due to cerbral apoplexy. The histological findings are quite specific and show myofibrillar degeneration with substitution of the normal striation by coarse sarcomeric transversal bands. The possible role of necrosis by catecholamines in sudden deaths and non-coronary myocardiosclerosis is emphasized.


[Bifascicular block: its course and prognostic significance in 52 cases followed-up for 15 years].

The authors report the results of a 15 years follow-up in a group of 52 patients with right bundle branch block and left anterior hemiblock. The incidence of complete AV block and sudden death in this group is compared to the one observed in a suitable control group. Among the bifascicular block group there was only one sudden death and one case of complete AV block. However, the groups did not differ statistically for mortality rates and average survival time. The authors suggest that bifascicular blocks should be regarded as a mere risk factor for complete AV block.


[Left anterior hemiblock in 4th phase in a patient with Prinzmetal's variant form of angina pectoris (author's transl)].

In the present study a patient with Prinzmetal's variant form of angina pectoris presenting intraventricular conduction disturbances (left anterior hemiblock in 4th phase) is reported.


[The vascularisation of the periorbita].

1. The periorbita is extensively vascularised from the side of the bone and the orbita. The vessels are interconnected so that the periost is not vascular boder area. 2. In the periorbita of the adult there is a dense capillary plexus which is less close-meshed on the inside. 3. Meandrian arteries and vascular loops exceptionally as long as 6 mm were found in the inner layers of the periorbita and in the adjacent layers of the fat connective tissue. 4. Flat membranes of fat connective tissue which are connected with the inner layer of the periorbita by long vascular streaks about on the inner surfaces of the periorbita. 5. On the outer surface of the periorbita venules 40 to 60 mum wide were found entering the periorbital bone. In the fissures area further meandering venules and venes were found which open in the cavernous sinus (in addition to the Vv. ophthalmicae) or enter the bordering bone. 6. N. and A. infraorbitalis run outside of the periorbita. 7. The nerves leave the periorbita by special channels and run through near and husk-like processes of the periorbita which accompany them up to the outer periost. In the bone they five off bone vessels.


[Does the weight of the brain depend on the body height?].

Brains of 1664 subjects (895 males and 769 females) aged from 20 to 89 years have been studied. The whole material being investigated was divided, within sex groups, into body-height classes and age classes. The class interval within the age classes was 10 years, that in height classes 5 cm. Mean arithmetics, standard deviations, standard error as well as coefficients of variation and correlation for respective classes have been calculated. It has been ascertained that the brain weight depends on the body height. In tall subjects no brains of extremely low absolute weight are encountered and, adversely, high brain weight is seldom met in short individuals. The body height also exerts certain influence upon the relative weight of the brain. More favourable proportion between the brain weight and the body length has been revealed in short subjects. Tall individuals are characterized by a low relative weight of the brain. It should be supposed that the spinal cord weight is higher in the latter subjects. The differences between the mean absolute weight of women's brains and that in men of the same age class are conditioned by the difference in the body length. A constant magnitude of difference in the mean brain weight in subjects of the same body height claims 100 g. The paper provides 2 enclosed tables representing obtained results for arithmetic mean of the absolute brain weight both in the age classes and body height classes. The differences between the mean weights of brains in women as well as in men are not significant. The coefficient of correlation between the brain weight and the body height is for men r male1 = 0.2008 for women r female1 = 0.2630, wherease the coefficient of regression for the brain weight is r male2 = 3.67 and r female2 = 3.906 respectively.


Studies on the morphology of the gills and gill arches with reference to the blood supply in Notopterus motopterus and Colisa fasciatus.

The morphology of the gills, with their blood supply have been described in Notopterus notopterus and Colisa fasciatus in some detail. Gills are curved and perforated on the dorsolateral and ventrolateral wall of the pharynx. The gills consist of 2 rows of filaments which are stacked one above the other to form a space. The gill filaments are smaller on both the ends and larger in middle. The gill filaments are of pink colour as they are supplied with blood. Gill rakers are large in size in Notopterus notopterus while they are small in Colisa fasciatus. 3 pairs of basibranchials are present in Notopterus notopterus which are covered by median membranous bony plate while 2 basibranchials are present in Colisa fasciatus. 3 pairs of hypobranchials are present in both fishes. 5 pairs of ceratobranchials are present in which Vth ceratobranchial bears teeth. 4 pairs of epibranchials are present. 3 pairs of pharyngobranchials are present in which the tip of the IVth pharyngobranchial bears minute teeth in Notopterus notopterus while in Colisa fasciatus IInd and IIIrd pharyngobranchial bear minute ones. One afferent branchial vessel is present in Notopterus notopterus and Colisa fasciatus in each gill like in other teleostean fishes. One efferent branchial vessel is present in each gill of Notopterus notopterus while in Colisa fasciatus 2 efferent are represented in each gill.


[Details of the muscular and nervous system in the anterior member of Sphenodon punctatus (reptiles-Rhynchocephales)].

Sphenodom is the only Lepidosaurian with an organisation of the nervous ways in the anterior member which constitute an important stage in tetrapod evoluation. Squamata are exceptions. The radial nerve is developed and possesses the most important part of the fibres for the dorsal muscles of the forearm. The flexor canal is separated into 2 voluminous trunks.


Hypertrophic changes in the ovarian chromaffin tissue (catechol storing tissue) under the influence of the pituitary and its theoretical implications in the teleost fish, Nandus nandus.

Evidence is presented in this work for the existence of a special chromaffin tissue in the ovary of the percoid teleost fish, Nandus and its hypertrophy under the influence of the pituitary extracts. The relationship between the pituitary and the ovary has been discussed from various viewpoints and the secretions of the ovarian chromaffin tissue and the interrenal are suggested as the possible mediators in the process of maturation and ovulation in the teleost fishes.


Marginal overdominance in Drosophila.

A reanalysis of Drosophila viability data was undertaken to determine the role of genotype-environment interactions in the maintenance of polymorphism. Between-replicate variances of viabilities in chromosomal homozygotes and heterozygotes with the same mean fitnesses were compared, with the expectation that if the heterozygote variance were on the average greater, conditional overdominance would be prevalent; if it were less, partial dominance would be prevalent; and if it were the same, marginal overdominance of the type considered by Wallace (1968) would be the prevalent type of variation. In fact, heterozygote variance was slightly less. The work of Dempster (1955) and of Gillespie and Langley (1974) is cited to show that this situation can still lead to balanced polymorphisms. Their general model for genetic variation in populations, consistent with the viability data, is reinforced.


Genetic drift in clines which are maintained by migration and natural selection.

Genetic drift will cause a migration-selection cline to wobble about its expected position. A rough linear approximation is developed, valid when local populations are large. This is used to calculate effects of genetic drift on clines in a stepping-stone model with abrupt and with gradual changes of selection coefficients at a single haploid locus. Among the quantities calculated are measures of slope, standardized variation of gene frequencies around their expected values, and correlation among neighboring populations with respect to deviations from the expected gene frequencies. These quantities appear to be primarily functions of Ns and Nm for a given pattern of selection. Computer simulation gives rough confirmation of these results. Standardized variances of gene frequencies and correlation of neighbors differ along the cline in the case of smooth changes in selection. In no case is pathological behavior of gene frequency deviations found near the boundaries of selective regions. Local behavior of gene frequences of nearby colonies is approximately predicted by a simple adaptation of the stepping-stone theory of Kimura and Weiss. Approximate measures of the lateral variation of the midpoint of a cline and the probability of non-monotonicity are also calculated and discussed.


Genetic drift in a cline.

A model is developed of genetic drift in a cline maintained by spatially varying natural selection and local dispersal of individuals. The model is analyzed by an approximation scheme which is valid for weak selection and small migration rates. The results, which are based on numerical iterations of the approximate equations, are that the cline is less steep than predicted on the basis of the deterministic theory but that for weak selection the correlation between random fluctuations in neighboring colonies is approximately the same as in models of migration and drift in the absence of selection.


Genetic heterozygosity in unreplicated bacteriophage lambda recombinants.

Bacteriophage crosses using density-labeled parents have been carried out under conditions restricting DNA synthesis. The parental material and genetic contributions to progeny manifesting recombination within a genetic interval sufficiently short to exhibit high negative interference have been examined. The unreplicated products of recombination isolated as phage particles appear to contain long continuous heteroduplex regions which are heterozygous for the closely linked markers. Recombination between closely linked markers seems to be the consequence of the removal of base-pair mismatches that are present within the heteroduplex regions. This localized reduction of heterozygosity within the heteroduplex regions that join the parental components of recombinant DNA molecules can account for high negative interference.


Mutant enrichment in the colonial alga, Eudorina elegans.

An enrichment procedure has been developed that results in at least a 200 X increase in mutation frequency in the colonial alga, Eudorina elegans. A period of nitrogen starvation followed by treatment with 8-azaguanine results in the death of wild-type cells and the maintenance of mutants. N'-nitro-N-nitro-soguanidine-induced acetate, p-aminobenzoic acid and reduced nitrogen requiring mutants have been isolated by this procedure.


Distribution among the chromosomes of Drosophila pseudoobscura of the genes governing the response to light.

Most strains of Drosophila pseudoobscura are neutral to light when tested in phototactic mazes. However, clear-cut photopositive and photogenative populations are obtained by selection over a series of generations. The genetic nature of the differences between the positive and negative populations has been studied in crosses in which the three large autosomes carried mutant markers. All chromosomes contain genes which influence the response to light. The third chromosome has the strongest effect, followed by the second, the X, and the fourth chromosomes. This seriation is not in proportion to the relative lengths of the chromosomes. Either the effective genes are not very numerous, or some of them exert stronger influences than others.


Diallel analysis of growth traits in mice.

Two replications of a complete diallel cross experiment were performed among four partially inbred lines of mice. These inbred lines originated from a random-bred ICR strain and were produced by 12 generations of full sibbing (F congruent to 92%). Individual body weight was recorded for each animal at 12, 21, 42 and 56 days of age. Body weight gain traits were examined for intervals 12-21, 21-42 and 42-56 days. Simultaneous least squares analyses of inbred and linecrossed groups were used. Sex differences were highly significant for all traits. Replicate differences were significant but made a small contribution to the total variation. Inbred lines differed greatly. Crosses showed growth trends similar to their contemporary maternal and paternal inbreds. Heterosis was highly significant for all traits except 21-day weight. Inbreds were heavier at 12 days of age, but linecrossed progeny were superior to inbreds for all postweaning weights. General combining ability was highly significant for 12- and 56-day weights and 21-42-day gain. Specific combining ability was highly significant for 21-day weight, 12-21- and 42-56-day gain. Significant maternal effects were found for all individual weights but not for 12-21- and 21-42-day gain. Residual reciprocal effects were significant for all traits. Estimated variances among linecrossed groups contained a large maternal component, a fluctuating additive genetic component and consistent non-additive genetic influence on all growth parameters measured.


Studies on the nature and function of polygenic loci in Drosophila. III. Veinlet modifiers having region-specific effects upon the vein pattern.

Polygenic modifiers affecting the expression of the mutant veinlet were studied to determine whether each acts specifically upon one vein or wing region or whether they affect the venation pattern in some general way. Selection experiments showed that the L4 vein can be modified independently of the L2 and L3 veins. Similarly, the L2 vein can be shortened while the L3 is selected to be longer, although there is some interdependence between the L2 and L3 veins. Assays of heterozygous whole chromosome effects show that different chromosomes are involved in the responses of separate veins, and one polygenic locus causing a decrease in L4 vein length has been isolated. Substitutions of whole chromosomes from selection lines into unselected backgrounds of non-homologous mutants demonstrate that the selected modifiers have the same qualitative effects upon other mutants having similar phenotypes. These results support the hypothesis that the polygenic modifiers affecting veinlet expression function independently of the veinlet locus, presumably by influencing common steps in the developmental processes leading to the formation of individual veins.


Natural selection for within-generation variance in offspring number II. Discrite haploid models.

In the classical model of genetic drift in population genetics theory, use is made of a hypothetical "infinite-gametic pool". If, instead, the gametic pool is determined by the random number of offspring per individual, a new form of natural selection acting on the variance in offspring number occurs. A diffusion model of this selection process is derived and some of its properties are explored. It is shown that, independent of the sampling scheme used, the diffusion equation has the drift coefficient M(p) = p(1-p) (mul--mu2 + sigma2e2--sigma2el) and the diffusion coefficient v(p) equals p(1-p) [psigma2e2 + (l--p)sigma2el]. It is also pointed out that the Direct Product Branching process model of genetic drift introduces a non-biological interaction between individuals and is thus inappropriate for modeling natural selection.


Genetic modification of recombination rate in Tribolium castaneum.

Asymmetrical responses were obtained in a replicated study of 15 generations of two-way selection for recombination rate between the ruby (rb) and jet (j) loci in Tribolium castaneum. Recombination rates in the two replicate high lines increased from an average of 0.22 in the base populations to an average of 0.42 at generation 15. Recombination rate pooled over the 15 generations of selection in each low line was significantly less than the control but there was no clear downward trend in response to selection for decreased recombination rate. The realized heritabilities were 0.16 +/- 0.03 and 0.17 +/- 0.02 in the two high lines, and were not significantly different from zero in the two low lines. Selection was based on crossing over in cis females only; however, rates measured in cis males after 12 generations showed the same response patterns as female rates. Similar response patterns were also determined for recombination measured in trans males and females at generation 18 following three generations of relaxed selection. The distribution of recombination rates measured in backcross beetles [(H X L) X H and (H X L) X L] at generation 12 indicated  polygenic control with those genes decreasing recombination rate being dominant. Detailed analysis of recombination rates in F1's produced by interline crosses at generation 15 confirmed the directional dominance findings. Under a polygenic model of recombination modifiers in which low recombination is dominant to high, average recombination rates will increase as inbreeding progresses, thus providing a mechanism for the production of new gene combinations in small populations.


Population genetics of euphydryas butterflies. I Genetic variation and the neutrality hypothesis.

Twenty-one populations of the checkerspot butterfly, Euphydryas editha, and ten populations of Euphydryas chalcedona were sampled for genetic variation at eight polymorphic enzyme loci. Both species possessed loci that were highly variable from population to population and loci that were virtually identical across all populations sampled. Our data indicate that the neutrality hypothesis is untenable for the loci studied, and therefore selection is indicated as the major factor responsible for producing these patterns. Thorough ecological work allowed gene flow to be ruled out (in almost all instances) as a factor maintaining similar gene frequencies across populations. The Lewontin-Krakauer test indicated magnitudes of heterogeneity among standardized variances of gene frequencies inconsistent with the neutrality hypothesis. The question of whether or not to correct this statistic for sample size is discussed. Observed equitability of gene frequencies of multiple allelic loci was found to be greater than that predicted under the neutrality hypothesis. Genetic differentiation persisting through two generations was found between the one pair of populations known to exchange significant numbers of individuals per generation. Two matrices of genetic distance between populations, based on the eight loci sampled, were found to be significantly correlated with a matrix of environmental distance, based on measures of fourteen environmental parameters. Correlations between gene frequencies and environmental parameters, results of multiple regression analysis, and results of principle component analysis showed strong patterns of association and of "explained" variation. The correlation analyses suggest which factors might be further investigated as proximate selective agents.


Growth of the pig: changes in body weight and body fluid compartments.

Body weight and the rate of change in TBW, ECW, and ICW were measured in 252 anesthetized pigs during the first 12 weeks after birth. After TBW was measured with 3H2O, 55 of the pigs were killed and TBW measured by desiccation. 3H2O overestimated TBW by 6.5% of body weight and 4.9% of fat-free wet weight, compared to desiccation (P less than 0.001); mean figures for 3H2O were 78.6 +/- 1.02% of body weight, and for desiccation, 72.1 +/- 0.45%; on a FFWW basis, 88.6 +/- 0.94% for 3H2O, and 83.7 +/- 0.13% for desiccation. TBW decreased significantly from 85.0% of body weight at birth (1.5 dg) to 75% at 5 kg (day 28) at a rate of ---3.2% body wt/kg body wt (P less than 0.001 from a zero rate). After that the rate of decrease was not different from zero: --0.117% body wt/kg body wt. ECW decreased significantly from 48% at birth to 35% at day 28 at a rate of --3.802% body wt/kg body wt (P less than 0.001 from a zero rate), and after day 28 the rate of decrease was not different from zero (--0.149% body wt/kg body wt) through week 12. ICW decreased, but not significantly, at a rate of --0.099% body wt/kg body wt. The changes in the rate of decrease in TBW and ECW coincided with weaning, and it was speculated that there was a direct relationship between the two events.


Growth characteristics of intact and hypophysectomized female hamsters. The effect of bovine growth hormone on hypophysectomized hamsters.

Post-weaning growth data, including body length, body weight, and several visceral organ weights, were collected for intact and hypophysectomized female hamsters aged 4 to 12 weeks. In addition, hypophysectomized hamsters were treated with bovine growth hormone for a period of six weeks and the effects examined.