Effect of soil microstructure on the relationship between the intensity of nitrification and soil moisture.

The relationship between the logarithm of nitrification intensity and moisture (expressed as pF) did not have an ideal linear course in structural soils in which pores of a certain size predominated. When increasing the incubation moisture it could be observed that the intensity of nitrification decreases at pF 3.5-4.4 and increases again on further increasing the incubation moisture. The detected anomaly was observed when applying moistures corresponding to peaks of derivation pF-curves. It can be assumed that the relationship between the intensity of nitrification and moisture is modified by soil microstructure; therefore, this relationship was studied in more detail in the present communication.


Purification and chemical characterization of antigens from sheep erythrocytes.

A mixture of glycoproteins and glycolipids was solubilized from sheep erythrocytes membranes under the effect of high ionic strength (2 M NaCl, 0.5 M Tris). Several antigenic fractions could be purified from the mixture using gel filtration on Sephadex G-200 and block electrophoresis on Pevikon C870; two fractions were found to raise antibodies in a primary reaction and these antibodies effectively sensitized erythrocytes to lysis by complement. The majority of other fractions elicited a weaker primary reaction which was detectable by both agglutination and haemolysis. The fraction, migrating fastest towards the cathode, elicited after immunization a formation of antibodies that could be detected almost exclusively by haemagglutination. The fraction, which elicited in the primary reaction a high titre of haemolytic antibodies, is composed of 72% proteins, 11% lipids and 15% saccharides.


Anatomy of the bony pelvis in parapithecid primates.

Four partial innominate bones, attributed to the parapithecid primates Parapithecus grangeri and Apidium phiomense, have recently been recovered from Oligocene deposits in the Fayum of Egypt. These fossils provide the first documentation of pelvic morphology for early anthropoids. In pelvic anatomy, parapithecids show definite similarities to higher primates rather than to prosimians, but cannot be clearly allied with any one extant group. Functionally, the fossils indicate quadrupedal or leaping habits rather than suspensory or bipedal behaviors.


Ecology of sympatric gibbons.

Ecological compatibility of sympatric gibbons is achieved by a difference in diet resulting from the effect of differing body sizes on the efficiency of harvesting different foods. In addition, differences in feeding apparatus and body size between the large and small species maximise the rate at which each can eat its most eaten food type. Parallels exist between the species array of gibbons and that of the African apes in the relationship between body size and ecology.


The effects of marijuana and alcohol usage on handwriting.

In this study the handwriting of twelve individuals under the influence of marijuana, alcohol, and a combination of the two was compared with control samples. It was found that marijuana smoking in moderate amounts had less effect on the handwriting of the individuals than did alcohol. It  was also found that when the two were combined the gross changes observed were probably due to the effect of the alcohol alone.


Tissue distribution of paraquat and diquat after oral administration in rats.

The differential distribution of paraquat and diquat in liver, kidney, lung, brain and heart has been studied after oral administration of the LD50 or LD50/2 to rats. Paraquat concentrations were highest in all organs at 24 hours; at this time concentrations in kidney and lung were 2 to 3 times higher than at 2 hours. In contrast, with the exception of kidney, tissue diquat concentrations were highest at 2 hours. There was severe lung damage at 24 hours after paraquat; diquat aid not produce severe lung lesions, but caused intestinal distention and diarrhea. These findings suggest that the difference in the toxicity of paraquat and diquat is related to their difference in tissue distribution and excretion.


On the estimation of lay-time by measurements of rigor mortis.

Measurements of the torgue to overcome rigor mortis on the legs of 101 male human cadavers at 5 degrees C are reported. There seems to exist an upper limit, which is dependent on the lay-time, above which no value occurs, but below which every value may be found. By calculating the 90% tolerance limits for lay-times grouped at 12-hourly intervals, this upper limit was found to decrease exponentially with lay-time. This may be used for the estimation of maximum lay-times from rigor mortis measurements.


Mortality statistics and autopsy: reliability of estimation of the mode of death in Copenhagen and a rural district of Sealand, Denmark.

During a one-year-period 1243 medicolegal deaths in the City of Copenhagen were autopsied after the mode of death was provisionally determined by an external medicolegal examination. In 35% of the cases an atuopsy was requested by the police. Without autopsy the mode of death would have been defined erroneously in 10.4% of all cases. This was particularly because of the difficulties in distinguishing between natural and non-natural death. In a rural district of Sealand in the autopsy rate in the same period was 14%. The rate among presumed natural deaths and accidents was particularly low (7% and 26%, respectively) against 23% and 77% in Copenhagen. This means a greater risk for errors in the estimated mode of death compared to Copenhage. This can also mean unequal rights by law. It is stressed that the police, in some rural areas, should be more liberal in requesting autopsies and at least increase them to the Copenhagen level of about 30%. The external medicolegal examination alone offers too little reliability, and it is stressed that such differences from one part of the country to another should not exist in a small well-organised country such as Denmark.


Assay of major and minor constituents of opium samples and studies of their origin.

Samples of opium from different countries as well as from various regions of India were studied for their morphine and codeine content by gas-liquid chromatography (GLC) as their silyl ethers in a single step. Non-phenolic alkaloids (thebaine, papaverine and narcotine) were extracted with dichloromethane for GLC after conversion of the meconic acid, morphine and codeine in opium into insoluble salts with lime water. The data on alkaloidal constituents have been utilised in a determination of the origin of seized opium samples in this preliminary study.


Visualization of dyes on thin-layer chromatography plates by means of the argon ion laser.

A simple method for the detection of nanogram amounts of dyes by means of their fluorescence in the visible region of the spectrum is described. To stimulate the fluorescence, an argon ion laser with principal lines of 514.5 nm and 488.0 nm is used. The sensitivity of the technique allows the visualization of an amount of dye smaller than could be detected with either visible absorbance or ultraviolet stimulated fluorescence.


A routine screening method for the major metabolite of methyl phenidate in the urine of greyhounds.

A qualitative urine screening procedure for the determination of the major urinary metabolite of methyl phenidate, alpha-phenyl-2-piperidine acetic acid (ritalinic acid), has been devised. The method involves a salting-out extraction, conversion of the metabolite to the parent drug using methanol-hydrogen chloride, and simultaneous two-column gas-chromatographic detection. Final confirmation of the presence of the drug is by two-system thin-layer chromatography. The method is suitable for the detection of illicitly administered methyl phenidate in greyhounds.


[The value of computer tomography in cerebrovascular accidents].

Computer-tomography investigations in 300 patients with cerebrovascular diseases are reported. The possibilities of computer-tomography for localization and follow-up of ischaemic attacks, intracerebral haematoma and subarachnoidal haemorrhages are described. Details of clinical and diagnostic importance of follow-up studies are presented together with examples of current pathological data.


[Diagnostic value of sonography (B-scan) in the upper abdominal region].

Minimal risk, easy applicability and high efficiency of ultrasonography, makes this method a valuable screening procedure in upper abdominal diagnostics. Case history, physical examination and laboratory diagnostics should be followed by ultrasound procedures as the first directed examination. This review intends to evaluate the efficacy of ultrasonography in different gastroenterological, oncological, hematological and angiological diseases of the upper abdomen.


[Osteonecrosis morbus Friedrich].

This report presents in detail the aseptic bone necrosis of the sternal end of the clavicle, the so-called M. Friedrich. When pain persists this relatively rare illness can be operated successfully.


[The epidemiology of breast neoplasms].

In an epidemiologic study the risks of breast cancer were assessed. An evaluation of the distribution of different tumor stages revealed that more information, in a larger sense more civilization, workship and the life in town makes women more curious and brings them to go sooner to the doctor. A lack of information correlates with more advanced stage of tumor. Significantly more childless women suffer of breast cancer, but a high number of children is no protection. In women with 3 and more children yet more advanced stages were seen. This may be interpreted by isolation, lack of information and distress. A significantly high number of breast cancer patients lost one of their parents by death before the age of 20, which confirms the psychosomatic theory of cancerogenesis (overcoming of the "Objektverlust" in early childhood). The incidence of depressive states before the onset of the disease is significantly higher. A reduced capacity to communicate was also observed. The exploration of sexual behaviour showed more frequently a reduced sexual activity correlated to cohabitarche, number of partners, alibidimia, unwanted pregnancy and masturbation. The ability of adequate sexual life was deeply disturbed by disease and therapy. Following primary therapy intelligent guidance of the patient may avoid occurrence of disturbances of sexual communication.


[Intraoperative specimen radiography in mamographically suspect, non palpable breast lesions. Experiences with the Faxitron unit].

The excision of mammographically suspect lesions of the breast is a problem, when these are non-palpable. Our experiences with intraoperative specimen radiography in 128 cases show that this method is very useful and appropriate to prove biopsy, especially if calcifications are present.


[Combined use of colposcopy and cytology].

The widespread view of the nearly absolute accuracy of cervical cytology in the diagnosis of severe dysplasia, carcinoma in situ and small invasive carcinoma is not correct. The rate of cytologically missed cases computed from the first smear only is more than 10%. Using cytology and colposcopy simultaneously in each single case, the errors of one method will be greatly compensated by the other. Presupposition for the compensation, however, is the independent indication for biopsy. In other words, biopsy must be done based on cytological findings, even when colposcopy is unsuspicious. On the other hand a colposcopically suspected and cytologically unsuspected case must also be biopsied. For the gynecological clinic as well as for the gynecological practice, the additional effort resulting from the combined method is easily realizable. For large screening programs, however, the colposcope can be used only as an instrument for target punch biopsies, indicated by pathologic smears.


[Early diagnosis of minimal cerebral dysfunction].

The aim of this retrospective study was to find indicators for an early diagnosis of the MCD-syndrome. Two groups of children (n = 18), with and without MCD at the age of five years were compared in view of medical history and early developmental data. The results suggest that illnesses of the mother before the birth of the child, risk factors in the course of the second half of pregnancy, as well as slight retardation of the child's early psychomotor development deserve particular attention in further studies.


[Screening for elevated creatine kinase activities for the early diagnosis of Duchenne muscular dystrophy].

A screening test for the determination of creatine kinase in a dry spot of the whole blood is used for the early identification of boys with Duchenne muscular dystrophy and girls with carrier properties of this hereditary disease. In the absence of an effective medical therapy, such screening leads to genetic counselling of the affected families with the purpose of avoiding the birth of further cases of Duchenne muscular dystrophy in the same families. The first results of a voluntary screening program in Germany are discussed.


[Radiological diagnosis of juvenile urination disorders].

Voiding disturbances and available radiologic examinations are reported. The importance of a correct analysis of the findings for the orthopedic, nephrologic and neurologic treatment is emphasized. Intravenous urograms and voiding cystogram are standard methods; recently ultrasound is also included in the management of children with neurogenic bladder.


[Diagnosis of neurologic and motor development in pre-school age children with minimal cerebral dysfunction].

An investigation on neurologic development of pre-school children does not yet clearly differentiate between children with minimal neuromotor disturbances and healthy ones. This is due to the lack of knowledge on brain function in normal children. Diagnostic problems on Minimal Brain Dysfunction are demonstrated by neurologic and motor examinations of 4 to 6 years old children.


[School medical findings in children of foreign workers].

A report on three investigations of the medical school-service on children from foreign countries is given. The foreign children are normally smaller than German children, but in relation to their stature more heavily built. Vaccination status and documentation are insufficient. Nevertheless, disturbances in organic functioning considered to be attributed to damages caused by our "society of civilization" are less prominent in foreign children. The necessary actions of improving the above mentioned situation are pointed out.


[Perinatal problems in breech presentation].

543 cases of breech presentation at the 1st Clinic of Obstetrics and Gynecology of the University of Vienna were investigated. The rate of Cesarean section showed during the last years a distinct increase (1969 to 1973: 16,6%, 1974 to 1977: 27,9%, 1977: 39,7%). A significant decrease of perinatal mortality in cases of breech presentation was not observed. On the other hand an increase in Cesarean sections diminishes the rate of perinatal mortality. In this publication the findings concerning the obstetrical results using abdominal or vaginal delivery are compared. No significant difference in perinatal mortality could be found. A better result could only be achieved in cases of mature infants. Our point of view is, that at present the indication of Cesarean section for cases of breech presentation should be very liberal. On the other hand vaginal delivery in some cases is still of importance.


[The "Wiesloch model". Sports therapy and intervening prevention].

For more than 4 years independent practitioners in the area of Wiesloch have led an intervening prophylactic treatment. Based on the recognition of heart and circulatory diseases, the program consists of sport with medical participation, a series of lectures and a course an dietary cooking. The results of 4 years show that there is success in normalising pathological risk factors.


[Organized myocardial infarct rehabilitation].

Rehabilitation of patients with myocardial (coronary) infarction during 1969/70 has been analyzed. Thereafter a therapeutic chain was organized with rapid transfer from the acute hospital to a rehabilitation clinic and further treatment in ambulatory coronary groups at residence. Participants and non-participants of ambulatory coronary groups from 1969 to 1976 were compared. The results concern the influence of organization, return to work, behavior of physicians and patients, risk factors and physical capacity. During the first three years under study the death rates of participants and non-participants of coronary groups were 1.3% per year. The small number of patients does not permit further interpretation.


[Pathogenesis of diabetic microangiopathy. Protein and basement membrane synthesis in isolated kidney glomeruli of diabetic and non-diabetic rats].

In incubation experiments with isolated glomeruli, an increased synthesis of protein and basement membranes was detected in streptozotocin-diabetic rats compared to metabolically healthy controls. Chemical analysis of isolated basement membranes and incorporation studies did not give any indication of enhanced hydroxylation of lysine in the diabetic membrane. Different glucose concentration in the incubation medium and insulin in vitro did not influence protein and basement membrane synthesis of non-diabetic glomeruli. On the other hand, in diabetic glomeruli the synthetic activity depends on glucose concentration. Insulin had a stimulatory effect on protein and basement membrane synthesis diminished at lower glucose concentration and did not inhibit the increased synthetic activity demonstrated at higher glucose concentration. Therefore, these results may be attributable to an energy deficit of incubated glomeruli and not to a lower glucose stimulation of synthesis. By treatment of diabetic rats with insulin in vivo the synthetic activity was not affected by brief normalization of blood sugar. Insulin treatment from the beginning of diabetes only lead to a normalization of protein synthesis in moderate metabolic control. On the other hand, a rise of basement membrane synthesis could only be prevented by strict metabolic control of the rats. These results show that basement membrane synthesis reacts more sensitively to the diabetic situation than overall protein synthesis. Insulin deficiency does not appear to be one of the factors directly influencing basement membrane synthesis.


[Experimental data on the pathogenesis of hypertensive encephalopathy].

Experimental investigations of the pial vessels during acutely drug-induced hypertension revealed diffuse or sausage-like distension of arterioles. Degree and type of reactions depended on blood pressure characteristics like steepness of increase, percentual increase, duration and peak value in this order. Such vasodistension is interpreted as mechanical overwhelming of smooth muscle force within the arteriolar wall during acute increase of intraluminal pressure. Clinical consequences of these results are: Vasodilators as well as sedatives with respiratory depression are contraindicated for patients with hypertensive crises. Data also make readily apparent, that short lasting acute hypertension may be fatal for patients with preexisting disturbances of blood-brain barrier function as it accelerates the development of brain edema.


[Treatment of alopecia areata with dinitrochlorobenzol (DNCB)].

Twenty-two patients with severe forms of alopecia areata were treated about 4--16 months with dinitrochlorobenzene (DNCB) dissolved in acetone. According to the patients' reactivity to DNCB the chemical was applied to the scalp in various concentration once a week. In 14 patients we observed the therapeutic effect of DNCB is unknown. Possibly it might be due to a change in the local immunoregulation. At the present time this treatment cannot be recommended for general use.


[Congenital hereditary hypotrychosis. Generalized autosomal dominant hypotrichosis with pili torti (hypotrichosis congenita hereditaria Marie Unna)].

Report on a family with numerous relatives in 7 generations suffering from hypotrichosis congenita hereditaria Marie Unna type. Personal investigations were possible in 29 persons. 19 of them presented the characteristic features of the syndrome with pili torti, generalized inborn hypotrichosis and a type of baldness resembling androgenetic alopecia.


[Masked urothel carcinoma of kidney pelvis and ureter].

Behind the typical signs of a staghorn calculus of a kidney and a ureterocele with calculi in x-ray a transitional cell carcinoma of the kidney and the whole ureter was found. The nonfunction of the organ was thought to be due to pyonephrosis. Four other cases reported in the literature are discussed.


[Perinatal mortality in twin pregnancies. Progress in perinatology].

162 twin pregnancies of the I. Univ.-Clinic of Gynecology and Obstetrics in Vienna between 1969 and 1977 have been analysed (1969--1974 = group I; 1975--1977 = group II). It was found that the main risk factors of twin gravidity are the high rate of babies born before term, the high rate of operative deliveries and the high rate of perinatal mortality. The twin pregnancy should be diagnozed early. This allows the possibility of prophylactic therapy of onset of labour before term. According to our observations the difference in mortality rates between the first and the second group is very high (13,3 in comparison to 5,5%). The management of twin pregnancies as a high risk delivery with continuous fetal monitoring has doubled the number of indications for caesarean section. The essential problem of twin pregnancy seems to be the high rate of premature deliveries. Therefore an adequate increase of the legal antenatal and postnatal maternal protection period is emphasized.


[The importance of dietary fibers, also objections against the uncritical use of statistics].

It is explained that mathematical-statistical evalutions in epidemiologic investigations are not principally significant for the causal relationship of different events. This holds true especially for multifactorial connexions. Statistics is an auxiliary science, which is able to point to potential relationships, however, it is not a proof by itself. Unequivocal coincidental relationships can be connected statistically significant. In general the diseases which are related to a lack of fibers in food are typical prosperity diseases (civilization diseases). The same diseases were related earlier to an overconsumption of sugar and they are found also in overnutrition. By selection (e.g. the relation between dietary fibers and coronary heart diseases) and by ignoring important facts the conclusions can be manipulated. It is possible to show in some cases that wrong conclusions are drawn on the basis of a purely mathematical-statistical evalution of nutritional relationships. Even wrong data (e.g. for ethanol consumptions) are evaluated for many years.


[The effect of benzbromaron in gout patients with limited kidney function].

The effect of the uricosuric drug benzdromarone was investigated in 15 patients with hyperuricemia and renal failure. Benzbromarone micron was given in a dose of 80 mg/day for 6 days. The mean serum uric acid level fell from 8.9 to 4.24 mg/100 ml. After therapy uric acid concentrations in serum returned to 5.2 mg/100 ml. Renal urate elimination was increased from 487 mg/day to 831.3 mg/day; after medication urate elimination decreased to the initial values. Uric acid clearance was increased by benzbromarone from 3.97 to 11.1 ml/min, and even after therapy a uric acid clearance of 6.4 ml/min could be measured. A decrease of uric acid reabsorption from 4.73 to 2.3 mg/min was observed, after medication the reabsorption rate increased to 3.42 mg/min.


[Profiles of polycyclic aromatic hydrocarbon metabolites after treatment with various inducers of microsomal rat liver monoxygenases (author's transl)].

The microsomal oxidation of 12 frequently occurring environmental polycyclic aromatic hydrocarbons after incubation with rat-liver microsomes has been studied and their metabolites characterized by means of gas-liquid chromatography/mass spectrometry. The method enables the detection and characterisation of phenols, diols, triols, and tetrols as trimethylsilyl ethers beside the original hydrocarbons. Moreover, the induction properties of some carcinogenic and non-carcinogenic hydrocarbons (benz[a]anthracene, pyrene, chrysene, benzo[a]-pyrene, benzo[e]pyrene, benzo[b]fluoranthene, benzo[j]fluoranthene, benzo[k]fluoranthene) have been studied. Except pyrene and benzo[e]pyrene, all compounds investigated significant but different induction factors. The relevance of the induction for an estimation of the biological effect of environmental polycyclic aromatic hydrocarbons is discussed.


[Synthesis of the fragments A1-,, A9-15 and A16-21 of ovine insulin A chain using the S-tert-butylmercapto residue for thiol protection (author's transl)].

The following paper describes the synthesis of the fragments A1-8, A9-15 and A16-21 of the ovine insulin A chain using the S-tert-butylmercapto residue for thiol protection. The synthesized fragments, which showed a good solubility in organic solvents, were partially deprotected with trifluoracetic acid and converted to the corresponding S-sulfonates quantitatively by oxidative sulfitolyses at pH 7.6.


[Synthesis of the fragments A1-8, A1-7, A9-15 and A8-15 of the chicken insulin A chain (author's transl)].

By coupling the peptide derivatives H-Cys(SBut)-Cys(SBut)-His-OMe(6-8 b) and H-Cys(SBut)-Cys(SBut)-OH(6-7b) respectively with Trt-Gly-Ile-Val-Glu(OBut)-Gln-OH(1-5a) the N-terminal sequences A1-8 and A1-7 of the chicken insulin A chain have been prepared. The sequence of A9-15 has been obtained by connecting Bpoc-Asn-Thr(But)-Cys(SBut)-OH (9-11c) and H-Ser(But)-Leu-Try(But)-Gln-OH (12-15). Acylation of the aminopeptidderivative 9-15b with Bpoc-N2H3 yielded fragment A8-15 (8-15).


Lectin-associated proteins from the seeds of Leguminosae.

The seeds of Pisum sativum (pea), Canavalia ensiformis, Vicia faba, Vicia sativa, and Ricinus communis were shown to contain proteins which are associated to the respective lectins (lectin binders). The lectin binders from Pisum sativum and Canavalia ensiformis were studied more closely. Both are single proteins not resembling the variety of membrane glycoproteins found in animals and plants which bind to lectins. The pea lectin binder is a tetrameric glycoprotein composed of identical subunits of the Mr 51 000. Its interaction with the lectin is abolished by acidic buffers or by glucose. The Concanavalin A binder, which does not contain sugar, is composed of one kind of subunit, Mr of 35 000. As in the case of the pea lectin binder, glucose and acid dissociate the lectin-lectin binder complex, but in contrast to the pea lectin binder low NaCl concentrations also cause this effect. During germination and growth, the Concanavalin A binder appears in the roots.


The specificity of sialytransferases using glycosylated lysozyme derivatives as substrates.

Galactose, lactose, N-acetylgalactosamine, N-acetylglucosamine and fibrinoglycopeptides were bound to lysozyme by different linkages. These glycosylated lysozymes were tested as N-acetylneuraminic acid acceptors using particular sialytransferase preparations from frog and bovine liver and from bovine and porcine submandibular glands. Desialylated fetuin served as reference compound. Galactose residues of desialo-fetuin and lysozyme-lactose are sialylated by all four sialytransferases tested, galactose bound to lysozyme via a phenylazo group is inactive with the enzyme from bovine submandibular gland, and galactose bound directly to lysozyme serves as substrate only for the frog liver sialytransferase. Lysozyme-phenylazo-N-acetylgalactosamine is active only with the sialytransferase from bovine sumbandibular gland. N-Acetylglucosamine derivatives of lysozyme are inactive with all sialytransferases tested. These observations are discussed in the light of the natural substrates for the sialytransferases investigated.


The amino acid sequence of goat beta-lactoglobulin.

The isolation of beta-lactoglobulin from milk of the goat is described. The purified protein was checked for purity and has been characterized by its gross composition and end groups. The native or the modified protein was then degraded by tryptic and cyanogen bromide cleavage. The cleavage products were isolated and sequenced in the sequenator using a Quadrol and propyne program. These data provide the complete sequence of beta-lactoglobulin of the goat. The results are discussed and compared particularly with bovine beta-lactoglobulin components AB. Some biological aspects are described.


Self-association of band-protein from human erythrocyte membranes in aqueous solutions.

Band 3, the main integral protein of the human erythrocyte membrane, was solubilized and purified in high concentrations of acetic acid. After removal of the organic solvent by dialysis, the self-association of the protein in aqueous solutions was studied by analytical ultracentrifugation. Sedimentation velocity and sedimentation equilibrium experiments clearly demonstrate that, under appropriate conditions of protein preparation, at protein concentrations c less than 200 micrograms/ml, ionic strengths 2 less than 10mM and pH values remote from the isoelectric pH of the protein, band 3 shows a monomer/dimer/tetramer-association equilibrium. With some preparations, as well as at higher values of c or I, hexamers and octamers contribute to the association equilibrium. The time needed for relaxation towards association equilibrium depends on the blood donor from whom the membranes were derived and varies between less than one minute and more than several hours. The results of analytical ultracentriguation, together with previously published data on the incorporation of band 3 into planar lipid bilayers, from chemical crosslinking and from electronmicroscopy suggest that band 3 will also show a monomer/dimer/tetramer-association equilibrium in the human erythrocyte membrane. This hypothesis contrasts the widely-held assumption that, in the membrane, band 3 is a stable dimer; however, it is consistent with nearly all known data on band 3-self-association.


Synthesis and biological activity of seventeen analogues of human insulin.

We synthesized seventeen analogues of human insulin, applying the principle of stepwise, selective formation of the disulphide bonds. Most of these analogues only differ from human insulin in the replacement of a single amino acid in positions 2, 5, 6, 7, 8 and 11 of the A chain and 5, 7, 13 and 16 of the B-chain. The influence of these modifications on the physicochemical properties of the analogues is discussed. Eight analogues could be crystallized. All the analogues produce the same biological effects as insulin, but differ markedly in their potency. In isolated fat cells in vitro, [HisA8]insulin showed a relative potency of 2.46 in stimulating glucose oxidation (human insulin = 1), whereas [D-CysA6,A11]insulin had a potency of only 0.00027. Very low potency was observed when IleA2 or the half-cystines A6, A7, A11 or B7 were modified. Replacement of the invariant GlnA5 by alanine only reduced potency slightly. All the analogues are full agonists. The effects of the analogues on glucose oxidation and lipolysis are correlated, supporting the view that they are mediated by a common receptor on the fat-cell membrane. Hypoglycaemic potencies in the rat were similar to potencies in vitro. As expected, no correlation was demonstrable between antiserum binding--measured in the radioimmunoassay--and biological activity. Several results of this investigation are difficult to reconcile with the current view regarding the structure-activity relationship of insulin which appears to require further refinement.


Studies on the involvement of lysosomes in estrogen action, II. Seasonal variation in the sedimentation patterns of endometrial lysosomes from prepuberal pigs.

The lysosomal population in endometrial cells of prepuberal pigs varies with the seasons. In summertime, lysosomes equilibrate in sucrose density gradients over a broad, domeshaped area from p = 1.21 g/cm3 to p = 1.15 g/cm3. The winter pattern is characterized by a major sharp peak of aggregated lysosomes at p = 1.20 g/cm3, visible as a "snowflurry"-like band. The two extremes are linked by transition patterns in spring and fall. The possible correlation of this phenomenon to the seasonal fluctuations in the level of soluble cytoplasmic estrogen receptor is discussed.


A spot fluorometer for protein determination.

An apparatus is described which allows spot fluorometry in situ for quantitative evaluation of protein spots stained with a fluorochrome (Hoechst 2495).


[The separation of the 4-[4-(dimethylamino) phenylazo]phenylthiohydantoin derivatives of leucine and isoleucine on polyamid thin-layer sheets in the picomol range (author's transl)].

The separation of the 4-[4-(dimethylamino)-phenylazo]phenylthiohydantoin derivatives of leucine and isoleucine has so far not been possible. In this paper a method is described which separates these two amino acid derivates in the picomol range on polyamide thin-layer sheets. The system (10% formic acid/ethanol 10:9) can be used either in one or in two dimensions.


[Determination of the interaction of oligopeptides and oligonucleotides by a chromatographic technique (author's transl)].

Homologues of deoxyriboadenylic and deoxyribothymidylic acid show different degrees of retardation in a pentalysine gel. This chromatographic retardation is directly related to the strength of the interaction between the oligonucleotides and the immobilized pentalysine peptide residues. The strength of this interaction is determined by the total negative charge of the oligonucleotide, and by the nature of the constituent base of the oligonucleotide. The specificity of the interaction depends upon differences in conformation of the oligonucleotides.


Structure-function studies on red pigment-concentrating hormone, II. The significance of the C-terminal tryptophan amide.

The significance of the C-terminal tryptophan residue of the red pigment-concentrating hormone (RPCH: Glu-Leu-Asn-Phe-Ser-Pro-Gly-Trp-NH2) regulating the blanching of the crustacean chromatophores has been investigated. RPCH and a number of analogues that differ only in the C-terminal part of the hormone, have been synthesized and assayed for biological activity on the shrimp Leander adspersus. It has been shown that the indole skeleton of tryptophan is an absolute requirement for the biological activity of the hormone. To provide maximum response the tryptophan must be blocked as the amide. The activity of synthetic [Tyr4]RPCH and adipokinetic hormone (AKH) purified from Schistocerca gregaria has been compared with the activity of synthetic RPCH.


Interrelations between uptake of [14C]orotic acid and labeling of UTP and RNA in rat and mouse liver after partial hepatectomy.

The uptake and utilization of [6-14C]orotic acid for UTP and RNA synthesis were studied in rat and mouse liver at 24 h after partial hepatectomy. Rat liver concentrated radioactivity relative to blood several-fold better than did mouse liver after both sham-operation and partial hepatectomy. The results showed that in mouse liver, contrary to rat liver, the orotic acid uptake was not increased after the partial hepatectomy. In rat liver, the precursor uptake and the labeling of UTP increased by about 75% whereas the specific radioactivity of RNA increased 2 to 3-fold after the operation, thus indicating an increased RNA synthesis. Mouse liver showed no increased [14C]orotic acid uptake or labeling of UTP or RNA at 24 h after partial hepatectomy.


Snake venom. The amino-acid sequence of the subunits of two reduced and S-carboxymethylated proteins (C8S2 and C9S3) from Dendroaspis angusticeps venom.

Two proteins (C8S2 and C9S3) were purified from Dendroaspis angusticeps venom. Whereas protein C8S2 comprises 124 amino acid residues, protein C9S3 contains 125 and each includes 16 half-cystines. After reduction and S-carboxymethylation of the proteins, the subunits were shown to contain 62--63 residues including 8 half-cystine residues. The complete sequences of the subunits have been elucidated. The sequences of the subunits of protein C8S2 and C9S3 are homologous to those of protein S2C4 from D. jamesoni kaimosae. Further, in all the subunits one of the structurally invariant amino acids, Tyr25, has been replaced by phenylalanine or asparagine and the Cys66 occurs in position 57. All the subunits apparently contain only two of the five functionally invariant residues of the neurotoxins, viz. Lys27 and Trp29. Mixtures of proteins S2C4, C8S2, C9S3 and angusticeps-type proteins showed a marked synergistic toxic effect.


Characterisation of protein synthesis in cell-free extracts from different mammalian cells by their sensitivity to inhibitors of polypeptide-chain initiation.

Plasma cells and reticulocytes are differentiated mammalian cell systems specialized in the synthesis of distinct proteins. To study whether a cell specificity of polypeptide-chain initiation in such cell systems can be detected through inhibitors, the sensitivity to several drugs interfering with initiation was compared in cell-free systems with S-30 extracts from these cells. The following was indicated by the experiments: (1) under the selected conditions, the different cell-free systems are comparable with respect to their activity of initiation, and that aurintricarboxylic acid inhibits mRNA binding to the ribosome in plasma cell tumours the same as in reticulocytes. (2) The sensitivity of translation of endogenous mRNAs in reticulocytes and plasma cell tumours to inhibitors of mRNA binding to the ribosome, aurintricarboxylic acid and the homopolynucleotides poly(U) and poly(A), are different. (3) Inhibition of the succeeding reactions of peptide-chain initiation by sodium fluoride and pactamycin was not selective. (4) In both cell systems translation of poly(U) is equally sensitive to aurintricarboxylic acid. (5) In extracts from TEPC 15 myeloma cells synthesis of immunoglobulin L-chains compared with that of the other myeloma proteins is partially resistant to aurintricarboxylic acid, whereas in reticulocytes, no differential sensitivities of individual proteins could be observed. The different susceptibility of the mRNA binding reaction in plasma cell tumours and reticulocytes suggests that the predominant mRNAs of these cells have different affinities to ribosome binding sites.


Taurocholate uptake by adult rat hepatocytes in primary culture.

Adult rat hepatocytes were cultured on Petri dishes for 25--30 h prior to measuring their ability to transport taurocholate. A rapid uptake of the bile acid (25 muM) was observed: about 20% was accumulated in the cells within 15 min. The taurocholate transport was saturable with an apparent Km of 28 +/- 10 muM and a maximal velocity V of 0.07 +/- 0.02 nmol/(micrograms DNA x min). Uptake was shown to be energy dependent as it was inhibited about 65% by antimycin A (20 micrograms/ml). The monohydroxylated bile acid taurolithocholate and the dihydroxylated taurochenodeoxycholate inhibited taurocholate transport to about 30 and 40% resp. of the control. The transport process was strongly dependent on sodium ions. It is concluded that the characteristics of taurocholate uptake into adult rat hepatocytes are very similar either in freshly prepared cells or in hepatocytes which are cultured on Petri dishes for 25--30 h.


Novel opioid peptides derived from casein (beta-casomorphins). I. Isolation from bovine casein peptone.

A material which displayed opioid activity in the guinea pig ileum longitudinal muscle-myenteric plexus preparation was extracted from an enzymatic casein digest into chloroform/methanol. The extract was roughly purified by adsorption/desorption procedures using charcoal and Amberlite XAD-2 resin as adsorbents. A high degree of purity was achieved by high-pressure liquid chromatography of the material on muBondapak C18 and mu-Porasil columns and finally by gel filtration chromatography on a Bio-Gel P-2 column. Several pronase-resistant compounds with opioid activity were obtained.


Novel opioid peptides derived from casein (beta-casomorphins). II. Structure of active components from bovine casein peptone.

Material with opioid activity had been isolated from an enzymatic casein digest. It was shown to contain a pure heptapeptide with the sequence Tyr-Pro-Phe-Pro-Gly-Pro-Ile. The identity between the opioid principle and the peptide was proven by the fact that chemical reagents or enzymes effecting one would effect the other. After carboxypeptidase Y digestion a pentapeptide, Tyr-Pro-Phe-Pro-Gly, could be isolated; this peptide showed a higher opioid activity than the heptapeptide. The opioid peptides were highly resistant towards proteolysis, even by pronase. The sequence of the hepatapeptide identified it as a fragment of bovine beta-casein. Therefore it was named beta-casomorphin.


Interactions of band 3-protein from human erythrocyte membranes with cholesterol and cholesterol analogues.

The interaction between cholesterol and band 3-protein from human erythrocyte membranes was studied by incorporating the solubilized protein into monolayers of cholesterol and related sterols at the air-water interface and measuring the changes in surface pressure which accompanied protein incorporation. The following results were obtained: 1) Band 3-protein shows a very strong interaction with cholesterol monolayers. Both apolar and polar bonds contribute to this interaction. 2) Steroids with a structure slightly different from that of cholesterol (especially with respect to the polar group and the side chain) in most cases show a reduced affinity for band 3-protein. Thus, the protein-sterol interaction is highly specific. It is assumed that the protein-cholesterol interaction can be subidivided into two parts: an unspecific one which results from contributions from several sterol molecules, and a specific one which is due to the high affinity binding of the protein and cholesterol. The structural element responsible for the high affinity interaction is assumed to be a sterol-binding niche on the surface of band 3-protein. The sterol is thought to be held in the niche by a hydrogen bond at its polar head and a variety of hydrophobic bonds along its ring system and side chain.


[Hemoglobins, XXVIII. Phosphate-protein-interaction, gene expression and function: the genetic and allosteric control of the oxygen affinity of the fetal blood (author's transl)].

This work describes possible molecular mechanisms concerning the control of oxygen affinity in fetal blood of mammalia. There is a genetic control of oxygen affinity through a fetal gene: at constant phosphate concentration (Hb less than P2-glycerate) in humans there is a hemoglobin with only five binding sites to 2,3-bisphosphoglycerate, resulting in an increased oxygen affinity. In several species (sheep, cattle, goat) with Met-Leu as the N-terminal group of the beta-chains, the 2,3-bisphosphoglycerate binding sites are deleted in positions beta 1 and beta 2, so that the regulation is phosphate-independent and thus providing a fetal hemoglobin with an increased oxygen affinity. The allosteric control is observed in pigs. In the postembryonal development "adult" hemoglobin with seven contacts (beta-chains) is demonstrated. The increased oxygen affinity is achieved here by a reduced biosynthesis of 2,3-bisphosphoglycerate (Hb greater than P2-glycerate) (Rapoport-Luebering-cycle). The functional control is discussed with respect to the ontogeny of the hemoglobins.


Modification of pyruvate kinase activity by proteins from chicken liver.

The partial purification of a protein fraction inhibiting pyruvate kinase isoenzymes is described. The fraction was isolated from the (NH4)2SO4 step of the purification procedure for pyruvate kinase isoenzymes from chicken liver (Eigenbrodt, E. & Schoner, W. (1977) Hoppe-Seyler's Z. Physiol. Chem. 358, 1033-1046) by extraction with 1N NaOH, acidification to pH 3, ethanol precipitation and chromatography of the supernatant on DEAE-cellulose. The inhibitor fraction was further purified by disc gel electrophoresis using a gel gradient from 10 to 25%; this procedure separated activating proteins from the inhibitor fraction. The inhibitor fraction inhibited the pyruvate kinase isoenzymes from chicken in the sequence of decreasing effect: M2 greater than L greater than M1. The inhibition was due to a decrease in the affinity for phosphoenolpyruvate. The inhibitor is stable against heating for 5 min in 1% sodium dodecyl sulfate at 100 degrees C; it is destroyed by pepsin digestion. The inhibitor fraction could be purified further only by dodecyl sulfate gel electrophoresis. This resulted in the separation of 2 inhibitors (Mr = 33,500 +/- 8500 and ca. 5000), an activator (Mr = 15,100 +/- 5200), and an unidentified protein (Mr = 27,000).


Synthesis of anomeric ethyl ketosides of 5-N-acetyl-D-neuraminic acid.

The syntheses of anomeric ethyl ketosides of 5-N-acetyl-D-neuraminic acid are described. The alpha-anomer prepared by a modified Koenigs-Knorr procedure starting from acetochloroneuraminic acid is quantitatively cleaved by Vibrio cholerae neuraminidase. Proton-catalyzed reaction of 5-N-acetyl-D-neuraminic acid with ethanol yields the beta-anomer.


Methylation of DNA in developing embryos of the sea urchin Psammechinus miliaris.

Embryos of the sea urchin Psammechinus miliaris have been labelled after fertilization with [6(-3)H]uridine and cultured in filtered sea water. 32-cell, blastula, gastrula and pluteus stages were harvested. The DNA from these embryos was purified and hydrolyzed and the nuclear bases were analyzed by means of high performance liquid chromatography. The ratios of 5-methylcytosine and cytosine demonstrate that the concentrations of 5-methylcytosine are essentially the same in the developmental stages examined (gamma = 95%), which contradicts the hypothesis that methylation of DNA plays a role in cell differentiation.


RNA synthesis in isolated bovine thyroid nuclei and nucleoli. alpha-Amanitin effect, a hint to the existence of a specific regulatory system.

DNA dependent RNA polymerase activities in isolated bovine thyroid nuclei and nucleoli have been studied. They retain their RNA synthetic activity for an extended period of time. This RNA synthetic activity is sensitive to actinomycin D and requires the presence of all four ribonucleoside triphosphates. The optimal conditions have been determined. Polyacrylamide gel electrophoresis reveals that the RNA synthesized has a size distribution ranging from 34S to 4S. The production of 18S-8S RNA is very sensitive to low concentrations of alpha-amanitin. However, in isolated bovine thyroid nuclei (not in nucleoli) this drug displays an effect on all RNA classes produced. The alpha-amanitin induced drastic decrease of [3H]-UMP incorporation in RNA of all sizes synthesized by isolated bovine thyroid nuclei is discussed.


Preparation and fractionation of membrane vesicles of thymocytes after osmotic cell disruption.

The isolation of plasma membranes is often accompanied with a loss of sensitivity to stimulatory agents (e.g. mitogens). Changes in the structure of the cell membranes after cell breakage have also been reported. Concerning this problem, "mild" cell disruption conditions were tested by using osmotic shock. Different membrane fractions were isolated, resulting in an enrichment of plasma membranes in one fraction. All fractions were characterized by plasma membrane marker enzymes (alkaline phosphatase, gamma-glutamyltransferases), the amount of cholesterol, the molar ratio of cholesterol and phospholipid, by dodecyl sulfate-gelectrophoresis and by electronmicroscopy. Total balance sheets of the fraction were made for each of the different biochemical parameters. The enriched plasma membranes resulting by using the described method had also lost the sensitivity to the stimulatory effect caused by mitogens such as Concanavalin A.


Paraganglion cells mimicking metastatic clear cell carcinoma.

Four patients are presented whom normal paraganglion cells simulated metastases of clear cell carcinomas of the urogenital tract. The cells, located within and near the lumbar sympathetic ganglia, were initially interpreted by the pathologist as metastatic, but a later examination showed them to be normal paraganglion cells. One ganglion was also examined electron microscopically. Normal ganglion cells were seen, as were cells with intracytoplasmic osmophilic neurosecretory granules, confirming their paraganglionic origin. Awareness of the appearance of paraganglion cells in some instances is essential to avoid their misinterpretation as metastases of clear cell carcinomas.


Quantitative analysis of histological parameters in giant cell lesions of the jaws and long bones.

Stereological techniques have been used to compare histological parameters of the giant cell component in 10 giant cell tumours of long bones, 10 central giant cell granulomas of the jaws and 10 peripheral giant cell granulomas of the jaws. The giant cell parameters investigated were: mean profile diameter, mean diameter, mean profile axial ratio, volume-to-surface ratios, nuclear numerical density, profile area and cell volume. Serial sections were cut at 4 micron from routinely fixed, paraffin-embedded specimens of each case and two sections, separated by 100 micron, were stained with haematoxylin and eosin. Random fields were photographed until at least 25 giant cell profiles had been recorded from each of the two sections of each lesion. The processed film was projected for point and intersect counting, the entire procedure ensuring double-blind assessment of the cases under investigation. In general there was no significant difference for any of the parameters between the giant cells of central and peripheral jaw lesions. There was a significant difference, however, between the giant cells of central jaw lesions and long bone tumours in respect of both nuclear numerical density and mean absolute cell volume.


Acute diarrheal infections in infants. I. Epidemiology, Treatment, and prospects for immunoprophylaxis.

Epidemiologic differences that appear to be geographic or climatic actually relate more closely to socioeconomics and sanitation. Regardless of etiology, the major management problems are those of dehydration and its sequelae. Progress toward development of antibacterial and antiviral vaccines is discussed. Next month the viral and bacterial agents that cause diarrheal infections will be reviewed.


Ring chromosome 6: case report and review of literature.

A ring chromosome 6 has been identified by GTG-banding in a male with microcephaly, growth retardation, seizures, epicanthus, hypertelorism, micrognathia, and other congenital anomalies. Cytogenetic studies indicate the instability of the ring chromosome. The most common findings in subjects with ring 6 include: profound to moderate mental retardation, microcephaly, prenatal growth failure, retarded bone age, epicanthal folds, flat nasal bridge, short neck, ears low-set or malformed, microphthalmia, and micrognathia. Linkage studies, including HLA, are consistent with reported maps of chromosome 6.


Sensitivity of Bloom's syndrome lymphocytes to ethyl methanesulfonate.

Ethyl methanesulfonate induced several times as many sister chromatid exchanges (SCE's) in lymphocytes from individuals affected with Bloom's syndrome as in lymphocytes from controls or heterozygotes. In cultures of cells from an individual with Bloom's syndrome who had two populations of lymphocytes circulating in his blood--'low' cells having normal spontaneous frequencies of SCE's and 'high' cells having elevated frequencies--only the high cells showed the increased sensitivity to ethyl methanesulfonate.


The genetic hypothesis for susceptibility to lepromatous leprosy.

Evidence for genetic influence of the host response to infection with Mycobacterium leprae is reviewed. A complex segregation analysis is performed on data for 91 families from Mactan, Philippines, in each of which at least one offspring developed lepromatous leprosy. The data are not found to be inconsistent with an autosomal recessive hypothesis for susceptibility to lepromatous leprosy. Heritability estimates in the range of 80% were calculated for sib-sib pairs under the multifactorial hypothesis for susceptibility. It is argued that the multifactorial hypothesis is more in keeping with available immunologic, epidemiologic, and demographic data than is the single gene hypothesis.


Prominent acrocentric chromosome satellites in child patients with mental retardation or psychiatric disorders; no IQ-satellite size correlation.

Child patients with mental retardation or psychiatric disorders were selected for the presence of prominent acrocentric chromosome satellites and highest or lowest IQ scores. No correlation was found between IQ scores, of which eleven were below IQ 54 and 16 above IQ 68, and the length of acrocentric short arm, satellite, stalk, or short arm material below the stalk. Conventional staining, quinacrine fluorescence, C banding and ammoniacal silver staining revealed the apparent same acrocentric short arm variation between patient and one parent in eight families studied. These findings suggest that the satellite variants were truly normal variants and not etiologically related to the developmental disorders seen in the patients.


The C4 system: formal and population genetics.

The present study shows that the C4 system as investigated by high voltage agarose gel electrophoresis is highly polymorphic. In a series of unrelated Norwegian adults, where C4 types have been ascertained through segregation in families, six different haplotypes have been found to occur with a frequency exceeding 1%. The genotype frequencies in the population fit expected Hardy-Weinberg distribution. In family material comprising 89 matings with 327 children the distribution of offspring is as expected according to autosomal codominant inheritance of haplotypes.


A third common allele in the transferrin system, TfC3, detected by isoelectric focusing.

Further genetic heterogeneity of the transferrin system (Tf) was revealed by prolonged isoelectric focusing (PAGIF) of human sera on polyacrylamide gels (pH 4--6.5). One of the two common subtypes of TfC, designated previously as TfC1, is split into TfC1 and the new subtype, TfC3. The gene product of TfC3 has an isoelectric point between C1 and C2. In our sample (n = 252) seven phenotypes, TfC1, C2-1, C2, C3-1, C3-2, C3, and C1B2, were observed and the following frequencies calculated: TfC1 = 0.795, TfC2 = 0.155, TfC3 = 0.042, and TfB2 = 0.008. Family studies (n = 25) indicate an autosomal codominant mode of inheritance. The six TfC phenotypes are present after treatment of sera with neuraminidase and can be revealed in part by PAGIF and subsequent immunofixation.


Gm, Am and Km immunoglobulin allotypes of two populations in Tunisia.

Gm, Am and Km allotypes were investigated in two Tunisian populations (236 samples from Mahdia and 142 samples from Sfax). These populations descend from immigrants and, therefore, the results were compared with those obtained in other populations living in the Near East and in North Africa. The subclass heavy chain allotypes G1m, G2m, G3m and A2m are inherited in fixed combinations. There were five main and four minor Gm-Am haplotypes that could be deduced from the phenotypes. This led to the conclusion that the populations studied are Caucasoids with some African admixture (about 10%) and a very low oriental contribution. Furthermore, there were 11 samples which showed 8 uncommon Gm-Am phenotypes. These could be explained by the assumption of five different uncommon Gm-Am haplotypes. Four of these may have arisen by equal crossing over of prevalent haplotypes. The fifth may be the result of unequal crossing over of prevalent haplotypes. The fifth may be the result of unequal crossing over, since it was proven, by family study, that more markers are transmitted together than are present in the prevalent haplotypes.


The rate of sister chromatid exchange in normal human bone marrow cells.

Sister chromatid exchange (SCE) was studied in bone marrow cells of six healthy individuals. Compared with peripheral blood lymphocytes, a remarkably low and constant rate of SCEs was observed, ranging from 3.64 to 4.65 per metaphase. The lower incidence of SCE can be related only partly to the higher contraction of bone marrow cells and the shorter exposure time to BUdR. Rather, a cell-specific phenomenon is suggested.


Ring chromosome 10 associated with multiple congenital malformations.

A 46,XY,r(10) karyotype was found in lymphocytes and skin fibroblasts cultured from a 8-month-old male showing multiple malformations and severe mental retardation. A comparison of the clinical features observed in cases in which a 10 ring was identified by means of banding techniques has been also attempted.


Interstitial deletion of the short arm of chromosome 2 in a moderately mentally retarded boy without gross clinical stigmata.

This report describes a moderately mentally retarded boy in whom there is interstitial deletion of the short arm of chromosome 2 (del(2)(p11p21)) associated with the presence in all cells of a small acentric fragment.


Multibranched chromosomes 1, 9, and 16 in a patient with combined IgA and IgE deficiency.

Instability of the centromeric region of chromosome 1 and multibranched configurations formed by different numbers and combinations of arms of chromosomes 1, 9, and 16 were found in cultured lymphocytes of a 12-year-old male with combined IgA and IgE deficiency. No chromosome abnormalities were found in fibroblast cultures from the patient or in blood cultures from his parents. A possible effect on the frequency of the abnormalities of the almost continuous antibiotic treatment received by the patient was found both in vivo and in vitro, but no abnormalities were found in blood cultures from control subjects who received similar treatment. Interphase association of chromosomes 1, 9, and 16 and a high frequency of interchanges among the centromeric regions of these chromosomes due to the presence of a fragile site is assumed to be the cause of the abnormalities.


Inverse maternal age effect in monosomy X.

The maternal age distribution of 45,X abortuses was significantly lower than that of chromosomally normal abortuses in two co-ordinated studies. One, carried out in Geneva, included 44 X-monosomic abortuses and the other, in Hiroshima, was based on 38 abortuses with a 45,X karyotype. It was deduced that 45,X conceptuses result from either non-disjunction during paternal meiosis or anaphase lag during meiosis or mitosis, and that anaphase lag is more frequent among young couples than among older ones.


An r(22)(p11 leads to q13) in a moderately mentally retarded girl.

An r(22) was detected in a 6-year-old female patient with growth retardation, IQ of 45, and a lower quotient for verbal performance. She presents some other minor anomalies. The break and fusion points probably were at p11 and q13.


The use of bone core biopsies for cytogenetic analysis.

Cultures of bone core specimens have proved satisfactory for cytogenetic analysis in patients from whom it was impossible to obtain a bone marrow aspirate, or in whose peripheral blood dividing myeloid cells were absent or insufficient in number. The quality of the metaphase chromosome is adequate for banding studies.


Reduced frequency of sister chromatid exchanges in human lymphocytes cultured with autologous serum.

The incidence of sister chromatid exchange (SCE) was determined in human lymphocytes cultured with fetal calf, human AB, and autologous serum. In each individual studied, cells grown in medium supplemented with fetal calf and human AB serum showed higher yields of SCE than those cultured with autologous serum. Increased concentration of fetal calf and human AB serum in the tissue culture medium resulted in elevated frequency of SCE. No such elevation in SCE frequency was observed with increased concentration of autologous serum. The results indicate the presence of extraneous SCE-inducing factors in fetal calf and human AB serum, the nature of which is not precisely known.


Structural differences in reciprocal translocations. Potential for a model of risk in Rcp.

Interchange segment sizes and the sizes of chromosome imbalance arising from the different modes of meiotic segregation were measured in a selected sample of 20 reciprocal translocations (Rep). The Rep were selected by two modes of ascertainment: (I) neonates with an unbalanced form of the translocation, and (II) couples with recurrent spontaneous abortions without evidence of full-term translocation aneuploid offspring. The measurements (% of haploid autosomal length: %HAL) were plotted as the observed or potential chromosomal imbalance with monosomy (abscissa) and trisomy (ordinate). It was found that (a) the interchange segments were larger in the spontaneous abortion Rcp, (b) that all of the imbalances observed in full-term neonates plotted close to the origin and to the left of the line joining 4% trisomy to 2% monosomy, and (c) the imbalances observed in the neonates in each individual Rcp were of the smallest size possible arising by any segregation mode. It was concluded that a major factor in the survival to term of aneuploid conceptuses is the size (proportion of genome) of the chromosome abnormality, irrespective of the origin of the chromosome regions. These results are discussed in relation to their use as a model to evaluate the risk of abnormal offspring in the progeny of translocation heterozygotes (the Chromosome Imbalance Size-Viability Model).


Detection of carriers and prenatal diagnosis for fucosidosis in Calabria.

A significant proportion of patients affected with fucosidosis have Italian ancestors. We assayed for alpha-fucosidase activity purified mononuclear cells and/or leukocytes obtained from 64 members of two large pedigrees from Calabria, in which seven children had been diagnosed as affected with fucosidosis. Of these 64 individuals, 22 were diagnosed as carriers, while the values for the remainder were within normal limits, indicating a clear bimodal distribution among individuals at risk. These data confirm that carrier detection for fucosidosis requires the measurement of alpha-L-fucosidase activity in purified mononuclear cells from peripheral blood. In addition, this program has made prenatal identification of an affected fetus possible.


Genetic load in an isolated tribal population of South India.

The Kota of Nilgiri Hills, Tamilnadu, are an isolated tribal population and occupy the lowest stratum in the local social hierarchy. They have developed an economic symbiotic relationship with other tribes of the Nilgiri Hills (e.g., Toda, Kurumba, Badaga), but have almost no social relationship with other communities, such as the Hindu and Muslim, communities, etc. The total population of the Kota is about 1200. Consanguineous marriages are highly favoured in this group. This paper presents data on prenatal, infant and adolescent mortality in relation to the degree of inbreeding. No perceptible difference has been found in mortality figures between consanguineous and non-consanguineous marriages. This may be due to the long history of inbreeding among the Kota. No case of visible congenital malformation has been noticed. The estimates of genetic load as revealed by inbreeding data indicate that genetic load in the Kota is low (perhaps about 1 lethal equivalent per gamete); it is also low in comparison with that in other Indian populations.


Serum albumin variants from populations of Andhra Pradesh, S. India.

1108 tribal and 1062 non-tribal individuals from three districts of Andhra Pradesh were examined for serum albumin variants. A slow-moving variant, identical to Albumin Kashmir was found in a single Muslim individual. Another new slow-moving variant, faster than Albumin Kashmir found in a single individual of a Koya Dora tribe is designated as Albumin Koya Dora.


G6PD deficiency with Gd(-)A like variant in a Chinese family from Cambodia.

A low rate value of G6PD was found in red blood cells from a Cambodian boy. Enzyme mapping was performed according to the WHO standard methods. G6PD presented all the characteristics of the A(-) variant encountered in the Negroes and behaved distinct from fast migrating enzymes described in China. No negro was in the ancestry of the mother.


Primary mesodermal dysgenesis of the cornea (Peters' anomaly) in two brothers.

In this report two brothers with a severe form of Peters' anomaly, i.e., primary dysgenesis mesodermalis of the cornea, are described. The third-degree consanguinity of the parents is an additional argument for the autosomal recessive inheritance of this condition.


Genetics and biochemistry of the phenylketonuria-present state.

Phenylketonuria is an autosomal recessive inherited disease caused by a disturbance in the phenylalanine hydroxylating system. Phenylalanine is converted to tyrosine by phenylalanine hydroxylase, which is located mainly in the liver. This enzyme needs the reduced cofactor tetrahydrobiopterin to be active. In phenylketonuria, low or zero enzyme activity is measured. Enzyme activity higher than 5% compared with that in normal controls is correlated to hyperphenylalaninemia. Dihydropteridine reductase regenerates the active cofactor. A defect in this enzyme or in the biosynthesis of the cofactor results in phenylketonuria which does not respond to dietary treatment because the biosynthesis of neurotransmitters is impaired.


Cytogenetic effects of inactivated influenza virus on male germ cells of mice.

Earlier we reported that both actively replicating (A2 and B) and live but nonreplicating (C) strains of influenza virus induced significant percentages of chromosome aberrations in the spermatogonia of the infected mice, suggesting that virus replication was not essential for the induction of chromosome aberrations. In the present communication, data are presented to demonstrate that the inoculation of formaldehyde-inactivated A2 Hong Kong influenza virus also induced higher percentages of chromosome aberrations in mouse spermatogonia than were seen in controls.


Partial translocation of NOR and its activity in a balanced carrier and in her cri-du-chat fetus.

Cytogenetic analysis of a balanced reciprocal translocation t(5;13) carrier and her unbalanced 5p- conceptus was carried out. This carrier mother had previously given birth to a child with cri-du-chat syndrome. Silver staining demonstrated the breakpoint on 13p within the nucleolus organizer region (NOR). In the carrier, the NORs both at the original site (13p) and at the translocated site (5p) were silver-stained, indicating that the rRNA genes at both sites were transcribed. The NOR at the derivative chromosome 5 was also silver-stained in the fetus.


Polymorphism of alanine aminotransferase (E.C.2.7.6.1): common and rare alleles.

Gene frequencies of common and rare GPT alleles derived from an investigation of 1139 unrelated, healthy individuals from southwestern Germany are given. GPT typing was performed by means of horizontal starch gel electrophoresis in a Tris-histidine x HCl buffer system. In addition, a new electrophoretic variant, GPT9, is described. The frequencies of the GPT alleles observed were calculated as: GPT1, 0.4987; GPT2, 0.4686; GPT1M, 0.022; GPT0, 0.005; GPT3, 0.0022; GPT4, 0.0025; GPT8, 0.0005; GPT9, 0.0005.


Genetic polymorphism of alpha-L-fucosidase in Brittany (France).

The authors studied the phenotypic distribution of alpha-L-fucosidase in a random sample of the population of the area of Rennes (France). The frequencies of Fu1 (0.64) and Fu2 (0.36) genes are significantly different from the frequencies observed in New York whites and blacks.


Incidence and characteristics of glucose-6-phosphate dehydrogenase variants in Japan.

A total of 3000 men living in Yamaguchi were screened for glucose-6-phosphate dehydrogenase (G6PD) deficiency using Beutler's spot test and three types of starch gel electrophoresis. These electrophoresis used a phosphate buffer system at pH 7.0, a TRIS-EDTA-borate buffer system at pH 8.6, and a TRIS-hydrochloride buffer system at pH 8.8. Fifteen G6PD-deficient variants were found at the rate of 0.5% and classified into four groups. As new variants, G6PD Konan, Kamiube, and Kiwa were identified. These three variants had a mild to moderate G6PD deficiency and were not associated with any clinical signs. G6PD Konan had fast electrophoretic mobility as compared with normal levels, G6PD Kiwa had slightly elevated electrophoretic mobility, and G6PD Kamiube had normal electrophoretic mobility. These three variants had normal levels of Km G6P, Km NADP, and Ki NADPH, normal utilizations of both 2-deoxy-G6P and deamino-NAPD, normal heat stability, and a normal pH curve. The other variant was G6PD Ube, which we had previously found in Yamaguchi (Nakashima et al., 1977). One boy with G6PD Ube was Korean.


Haplotype analysis of the linkage group HLA-A: HLA-B : Bf in Japanese.

Three hundred four HLA-A : HLA-B : Bf haplotypes of the Japanese population as deduced by family analysis are described. Several linkage disequilibriums were observed in the following two-factor haplotypes: HLA-A and HLA-B, HLA-A and Bf, and HLA-B and Bf. Positive linkage disequilibriums between HLA-A and HLA-B noted in the present study seem to be Japanese specific when compared with the results obtained from other ethnic groups reported so far. The striking finding is that three HLA-B : Bf haplotypes, namely B12-BfF, B15-BfF, and B7-BfS, are common and show linkage disequilibrium in both Japanese and European Caucasians (German). This finding not only confirms the proximity of the HLA-B and Bf loci but also suggests that the interaction of the two genes plays an important role in the maintenance of stable linkage disequilibrium.


Deficiency of arginine and lysine causes increase in the frequency of sister chromatid exchanges.

An increase in the rate of sister chromatid exchanges (SCE) was found when V79 Chinese hamster cells were exposed to increasingly severe degrees of arginine and lysine deficiency. The data suggest a possible function of chromosomal proteins, and of histones in particular, in the maintenance of the low normal rate of SCE.


Mitochondrial malic enzyme (E.C.1.1.1.40) in human leukocytes: formal genetics and population genetics.

Mitochondrial malic enzyme MEM (E.C. 1.1.1.40) is present in human leukocytes; the polymorphism of MEM thus can be easily demonstrated using routine starch gel electrophoresis. Data on formal genetics are given. The gene frequency of MEM1 was estimated to be 0.67 +/- 0.02.


Human mitochondrial glutamic-oxaloacetic-transaminase, GOTM: formal genetics.

The polymorphism of human GOTM was investigated in red blood cells by means of routine starch gel electrophoresis. The formal model of two common alleles, GOTM1 and GOTM2, at an autosomal locus GOTM was confirmed by examination of 640 mother-child pairs. The frequency of GOTM1 in this sample from southwestern Germany was calculated to be 0.981 +/- 0.003.


A new case of partial trisomy 15q-.

Partial trisomy 15 was observed in a newborn with malformations of the head and extremities. A t(5;15) translocation was found in the mother and maternal grandfather.


Prenatal diagnosis of a 13p+ karyotype.

Chromosome 13p+ was identified in an amniotic cell culture. The variant has been inherited for three generations without adverse effect.


A low frequency mosaicism for monosomy 21 in a live born female.

Monosomy 21, whether homogeneous or as a mosaicism, is very uncommon. We report here a 3-month-old white female with a low degree of monosomy 21 in the blood karyotype (6.5%, 110 cells counted) but not in the skin fibroblasts, which contained only the normal chromosome complement. The patient's physical features included microcephaly with frontal slanting; prominent occiput; ridge-shaped sutures; agenesis of the corpus callosum; large, prominent ears; high and narrow palate; micrognathia; tetralogy of Fallot; crowded toes; and dry, thick skin with very little subcutaneous tissue. The case is discussed in light of the suggested clinical features of the "monosomy 21" syndrome and the possible implications of such a low-grade mosaicism in prenatal diagnosis.


A pachytene chromomere map of chromosome 10.

A pachytene chromomere map of bivalent 10 is presented. Recent results from high-resolution metaphase banding document a similar pattern of intrachromosomal differentiation.


Heteromorphic X chromosomes in 46,XX males?

This paper reports an attempt to determine whether the short arm of one of the X chromosomes in XX males is longer than normal. In a blind study comparing coded photomicrographs of 15 G-banded mitoses from each of five XX males and five control females, the results were ambiguous and somewhat contradictory, but gave the impression of, or were compatible with, an XXp+ phenomenon in at least two of the five XX males. Measurements of the X chromosomes from the above cells and, in addition, from 15 mitoses from each of six XXY males, failed to disclose any XXp+ phenomenon. Statistical analysis indicated that in the five XX males there was no difference in the lengths of the two Xp arms. The reasons for the apparent discrepancy between the results of ocular inspection and measurement are discussed. The putative heteromorphism might be an alteration in shape, staining intensity, or position of bands, neither of which necessarily leads to an increase in length. We conclude that our results do not indicate any XXp+ phenomenon in the five XX males tested. However, the presence or absence of XXp+ is not in itself evidence for or against interchange betweenthe X and Y in the paternal meiosis. Our results emphasize that the etiology of XX males is likely to be heterogeneous.


Interstitial deletion of chromosome 13 and associated congenital anomalies.

An interstitial deletion of chromosome 13 with breakpoints at 13q22 and 13q32 is presented. The clinical findings associated with this deletion are discussed in relation to the correlations of specific chromosomal bands with constellations of congenital defects as described by Niebuhr and Ottosen (1973), Niebuhr (1977). Lewandowski and Yunis (1975), and Noel et al. (1976).


Trisomy 9p resulting from de novo 9/15 translocation and a 9p isochromosome.

Trisomy 9p with de novo 9/15 translocation and 9p isochromosome was observed in a mentally defective boy with typical clinical features for this syndrome. This chromosomal aberration is probably caused by the pericentric inversion of chromosome 9 of the patient's father.


Expression of esterase D and other gene markers in trisomy 13.

A gene dosage effect was observed for esterase D in a patient with trisomy 13. Isoelectric focusing of hemoglobin from the same patient showed three unidentified bands. A gene dosage effect for several other enzymes was not observed in two other patients with trisomy 13.


Distribution of HLA A, B and C antigens in an Australian population.

HLA A and B antigens have been determined in 2740 adult responders to a population health survey in Busselton, Western Australia. HLA A, B and C antigens have been determined in 481 schoolchildren. The antigen frequencies are generally close to those obtained elsewhere for subjects of British origin, but there are some differences from the frequencies found in North American Caucasians. The frequencies were not affected by the inclusion of genetically related individuals in the sample. Seventeen HLA A-B haplotypes, six A-C haplotypes and six B-C haplotypes had frequencies above 1%. A total of 1071 distinct phenotypes were identified out of the 5069 which are theoretically possible for the HLA A-B model used in the study. The most frequent phenotype was A2, B12 which occurred in 2.5% of the sample.


De novo 13q paracentric inversion in a boy with cleft palate and mental retardation.

A paracentric inversion in chromosome 13, inv(13)(q12q22), is described in a boy with mild mental retardation and multiple minor anomalies. Bromodeoxyuridine-late replication studies showed no changes in the replication pattern of bands in the abnormal chromosome 13. The relation between the proband's phenotype and his inv(13) is unclear.


Tandem duplication (5q13 to 22) in a mentally deficient girl.

A tandem duplication of 5q13 to 5q22, visualized using GTG- and RBA-banding, was found in a mentally defective girl with craniofacial dysmorphy.


Heterozygous expression in 3-M slender-boned nanism.

A 15-year-old girl affected by autosomal recessive 3-M slender boned nanism (3-MSBN) was studied. The clinically normal parents, two other obligate and two probable heterozygotes for the 3-MSBN gene from an unrelated family were radiologically investigated. All except one probably heterozygote showed mild features of the 3-MSBN, mainly bone slenderness and prominent talus. These findings are interpreted as demonstrative of the heterozygotic expression of the 3-MSBN gene.


Urinary pepsinogen isozymes: a highly polymorphic locus in man.

A genetic analysis of human urinary pepsinogen isozymes is presented. Nine discrete phenotypes were identified in a population survey of 215 unrelated Caucasian individuals. The phenotypes were characterized by differences among the staining intensities of the activated group I pepsinogens, Pg 5, Pg 4, Pg 3, and Pg 2. The genetic studies demonstrated that the codominant expression of four alleles, PgA, PgB, PgC and PgD, at a single genetic locus determined the nine phenotypes identified. Linkage analysis excluded close linkage of the Pg locus with the chromosome 6 markers HLA, GLO1, and Bf.


Hemoglobin G San José [beta 2 7 (A4) Glu to Gly alpha 2], beta thalassemia, and alpha thalassemia in a Sicilian family.

A 3-year-old child of Sicilian origin was found to have a severe form of Cooley's anemia. Investigations were extended to other members of her family. In three, a rare beta-chain structural Hb variant, Hb G San José [beta 7 (A4) Glu to Gly], was observed: in the father of the porposita heterozygosity for the abnormal Hb was found to be coexistent with beta o thalassemia; two sisters had lowered MCV and MCH values and levels of the abnormal Hb significantly lower than in other heterozygotes for Hb G San José. The alpha-chain/total beta-chain synthesis ratios suggest an alpha-thalassemic-like effect. Their mother had lowered MCV and MCH values, an Hb A2 level in the upper limit of the normal range, and a balanced alpha-chain/beta-chain synthesis ratio. Therefore, the possibility of coexistence of an alpha thalassemia trait with a beta thalassemia trait in the mother of the proposita and with Hb G San José heterozygosity in the two sisters who had lowered levels of abnormal Hb is discussed.


Complex segregation analysis of diabetes mellitus.

Complex segregation analysis was applied to a sample of 12,293 nuclear families each with at least 1 diabetic patient. The families were divided into two groups depending on the proband's treatment: insulin-dependent (IDG) and insulin-independent (IIG). Heterogeneity analysis has revealed a highly significant difference in the IIG group when families were divided into different mating types. The higher recurrence risk was found in the group with affected mothers. Also evidence for a major recessive gene was found in the IGG group, while it was not possible to distinguish between the hypothesis for absence of a major locus and absence of polygenic inheritance in the IDG group. Risks to develop the disease were calculated for a few typical situations.


Joint distribution of z transformations estimated from the same sample.

When correlations are estimated by maximum likelihood method from the same sample, Fisher's z transformations of the correlations (as well as the sample correlations themselves) are shown to be asymptotically distributed as multivariate normal with appropriate mean and dispersion matrix.


Hereditary essential myoclonus.

A new family with the rate condition of hereditary essential myoclonus (HEM) and the literature on HEM are presented. Some of these cases may previously have been reported under the title of Friedreich's paramylclonus multiplex. The present family, which is number 13 in the literature and in which 9 members in three generations had this benign disorder, is described. The diagnostic criteria have been tabulated.


HLA-GLO linkage analysis in 57 informative families.

The HLA-GLO linkage relationship was investigated among 37 single backcross families with 97 children and 20 double intercross families with 51 children. For the total number of families the value Zmas = 14.600 for theta = 0.060 was found. A total of 11 cross-overs between HLA and GLO were found leading to a recombination fraction of 0.067. 221 unrelated haplotypes were examined for linkage disequilibrium between HLA and GLO, and no disequilibrium was found.


Hydroxyproline metabolism in two sisters with hydroxyprolinemia.

Hydroxyproline metabolism was evaluated in two sisters with hydroxyprolinemia and their mother. 33 and 21% of an oral hydroxyproline load (200 mg/kg) was excreted by the sisters, 5.4% by the mother, and 1.3% by normal subjects. Plasma and erythrocyte values in the sisters and their mother were elevated, indicating that extra- and intracellular hydroxyproline pools were increased. Analysis for urinary glycolate and oxalate (metabolic products of hydroxyproline) showed no increased excretion by the two sisters, although the mother's excretion was normal. A deficiency of hydroxyproline oxidase in the two sisters was indicated by the lack of delta 1-pyrroline-3-hydroxy-5-carboxylic acid excretion.


A familial extra small marker autosome in persons with normal phenotype.

The propositus was referred because of sterility and oligospermia. His karyotype was 45, XY, t(13q14q). His father was dead; his mother and the only brother, who was fertile, both had 47 chromosomes, but a normal phenotype and normal intelligence. The additional chromosome was three quarters the size of a G chromosome and had satellites on the short and long arms.


Phytohemagglutinin effects on cultured human neural crest tumors.

Two human melanoma lines, RPMI-7931 and HS-294, respond to mitogenic stimulation by PHA. A dose-response curve for these lines can be demonstrated with maximal stimulation at 16 and 32 micrograms per ml and inhibition at 75 to 250 micrograms per ml PHA. The mitogenic effects of PHA were inhibited by N-acetyl-D-galactosamine. Neuroblastoma cells also exhibits a similar but less important dose-response to PHA. These data indicate that human melanomas and neuroblastomas may have PHA receptors or mechanisms for mitogenic stimulation which are analogous to those observed with normal lymphocytes.


Culture of adult rat lung cells: benzo(a)pyrene metabolism and mutagenesis.

A method is described for obtaining and culturing large numbers of lung cells from normal adult male rats. The lungs were perfused in situ to remove blood cells and then perfused via the trachea with a trypsin-collagenase solution to initiate tissue digestion. The tissue was further digested in the enzyme solution and approximately 2 X 10(8) viable lung cells were obtained per animal. Primary cultures contained a mixed cell population. Through eight subcultures about 70% of the cell population possessed an epithelial-like morphology, whereas the remaining 30% was fibroblast-like. Three clones of epithelial-like cells were isolated at the fourth subculture. The mass culture lung cells and the epithelial-like clone that was studied retained a normal karyotype and did not grow in soft agar. Both the mass culture cells and the epithelial clone metabolized the lung carcinogen benzo(a)pyrene (BP) to water-soluble products. Furthermore, the mass culture lung cells metabolized BP to intermediate(s) which mutated Chinese hamster V79 cells from ouabain sensitivity to ouabain resistance. These lung cell cultures have potential use in cell transformation, mutation and carcinogen metabolism studies.


Proliferation and morphology of chick embryo cells cultured in the presence of horse serum and hemoglobin.

We have shown previously that hemoglobin greatly stimulates chick embryo cell proliferation in Eagle's minimal essential medium supplemented with horse serum. In the present study we compared the effects of horse serum plus 10 micrometers hemoglobin to those of fetal bovine serum on subcultures of chick embryo cells serially propagated at high cell densities. The cells became elongated in the presence of fetal bovine serum and their rate of proliferation progressively decreased, whereas they became polygonal in the presence of horse serum plus hemoglobin and proliferated well in successive cell passages. The polygonal cells obtained in the presence of horse serum plus hemoglobin rapidly elongated if cultured at low cell densities in the presence of fetal bovine serum, but, in contrast, elongated cells did not yield polygonal cells if cultured at low densities in the presence of horse serum plus hemoglobin. It is possible that the polygonal and elongated cells are undifferentiated cells and differentiating myogenic cells, respectively.


Isolation and characterization of fibroblasts obtained by pulmonary lavage of human subjects.

Cells that possess the morphology and collagen synthetic capacity of fibroblasts were recovered by bronchofiberscopic subsegmental pulmonary lavage from patients with pulmonary fibrosis, from patients with miscellaneous nonfibrotic lung diseases and from healthy volunteers. Lavage cells were placed in tissue culture, observed for 2 to 6 weeks, and compared with human lavage pulmonary alveolar macrophages (PAM), WI-38 and IMR-90 human fetal lung fibroblasts, and adult lung tissue fibroblasts (CLAC-76). Lavage fibroblsts (LF) were identified as proliferating clones in monolayers of nonproliferating PAM and could be subcultured repeatedly. Fibroglasts were propagated from 28 of the 92 lavage specimens cultured. Time-lapse cinematography showed similar distributions of interdivision times for LF, CLAC-76 and WI-38, but the LF and CLAC-76 lines had slower mean migration rates than the fetal line. Light, scanning, and transmission electron microscopy of LF showed attenuated spindle-shaped cells with interdigitating filopodia, flat surfaces with few microvilli, and containing numerous cytoplasmic polyribosomes and rough endoplasmic reticulum. Extracellular fibrils with the appearance of collagen were seen. Collagen synthesis by LF was measured as 3.9% to 4.9% of the cell-associated protein sensitive to bacterial collagenase. This protein was rich in hydroxyproline, and had an electrophoretic migration pattern identical to known collagen. LF did not contain lysozyme although this enzyme was abundant in fresh and 1-week cultured PAM. Thus LF were similar to human fetal and adult lung tissue fibroblasts in their morphology, tissue culture characteristics, constitutive enzymes and collagen synthetic properties but were distinctly different from PAM.


Growth characteristics of skin fibroblasts and 3T3 cells entrapped by polymerizing fibrin.

Skin fibroblasts as well as 3T3 cells were cultured after entrapping freshly prepared cells in medium containing polymerizing fibrin. In contrast to cells grown on plastic substratum, fibrin-clot-cultured cells became highly elongated forming strands of cells. The strands interconnected by lateral cellular protrusions so that horizontal networks of cells were present throughout the clots. Cell growth as well as stretching were dependent upon the concentrations of fibrin. Highest growth rates were obtained with low fibrin concentrations (0.3 mg fibrinogen per ml). As shown by deprivation experiments nutritional limitations appear to be responsible for differences in growth rates observed in fibrin clots of higher density. In this system the fibrin meshwork serves as substratum for adhesion, elongation and multiplication of fibroblasts. The method makes it possible to study single cells in culture and the effects of persistent microenvironmental influences.


Localization of vtiamin A by autofluorescence during induced metaplastic changes in cultures of skin.

A technique was devised for following the uptake and location of vitamin A in organ cultures. Explants of 12- and 13-day embryonic mouse upper lip skin were grown for 3, 6 or 9 days in biological medium to which was added 0, 4.1 or 6.9 micrograms per ml of retinyl acetate. This form of vitamin A caused glandular morphogenesis of vibrissa follicles, and keratinization in epidermis and follicles was completely suppressed in 12-day explants and partially suppressed in 13-day explants. Frozen sections at 16 microns showed the white, nonfading fluorescence of keratin and the green, rapidly-fading fluorescence due to vitamin A which was captured by high-speed photography. Although more concentrated within lipid droplets in the dermis, the vitamin penetrated both the epidermis and the hair follicles. The ability to obtain permanent photographic records of the fading fluorescence makes this a useful method for analyzing vitamin A distribution as well as keratin distribution.


Effect of horse serum on neural cell differentiation in tissue culture.

The effects of various concentrations of horse serum on dissociated mouse glial precursor cells in colony cultures were evaluated. High concentrations (20% or more) favored cell attachment but inhibited cell proliferation and differentiation, whereas lower concentrations (5% to 10%) favored cell proliferation and differentiation. In fetal bovine serum the cells did not attach to culture surfaces to the same degree nor did they achieve the same level of differentiation as in corresponding concentrations of horse serum.


Inhibition of complement-dependent rosette formation after lymphocyte incubation with fatty acids.

Overnight incubation of lymphocytes with certain fatty acids bound to albumin has previously been shown to modify the fatty acid composition of cellular phosphoglycerides without impairing viability, and also to inhibit mitogen and antigen-induced lymphocyte transformation. In the present study, incubation of human lymphocytes with palmitic acid or with linoleic acid impaired their capacity to form complement-dependent rosettes but had no effect on either spontaneous rosette formation with sheep erythrocytes or Fc(gamma)-dependent rosette formation. With mouse lymphocytes, complement-dependent rosette formation was suppressed only following incubation with linoleic acid and there was no effect on Fc(gamma)-dependent rosette formation. Although the mechanism of these effects remains to be elucidated the results suggest that control of membrane lipid composition may modify the important in vivo immunological functions which involve complement receptors.


Radioassays of blood group M, N and T (Thomsen-Friedenreich) antigens.

Radioassays employing the double-antibody or Farr techniques were developed for the M, N and T antigens. Blood group glycoproteins were isolated by butanol extraction of red cell stroma and iodinated by the chloramine-T technique. The final purity of glycoprotein was over 75% as judged by radioimmunoassay (RIA). T activation of glycoprotein was obtained with neuraminidase. A specific RIA was obtained for the M antigen and was sensitive to approximately 10 ng of glycoprotein or glycopeptide. In the RIA system rabbit anti-M displayed a higher affinity for M glycoprotein than for M glycopeptide. A RIA that was entirely specific for the N antigen, could not be obtained. A radioassay, obtained for the T antigen with peanut agglutinin in the Farr technique, was sensitive to approximately 100 ng of T antigen and was readily inhibitable by monosaccharides. A RIA, obtained for the T antigen with rabbit anti-T, was entirely specific and sensitive to approximately 1 ng of T activated glycoprotein or glycopeptide but was not inhibitable by monosaccharides.


Post-translational modification of rat immunoglobulins synthesized in the Xenopus oocyte translation system.

The post-translational modification of rat immunoglobulin synthesised in Xenopus laevis oocytes was studied. The major products of translation of rat spleen poly-(A) containing mRNA were found to be assembled 7S immunoglobulin molecules indicating extensive modification of primary translation products. The possibility that these immunoglobulin molecules might include antibodies of defined specificity was investigated using spleen mRNA from rats hyperimmunized with ferritin and keyhole limpet haemocyanin. The presence of antibodies to immunizing antigen in oocyte translation products was determined by affinity chromatography on Sepharose-antigen columns and the synthesis of Sepharose-antigen binding antibodies was observed, equivalent to 2.5-3% of total immunoglobulins. The oocyte produced antibodies were of the same immunoglobulin class as the circulating antibodies from the immunized rats.


Suppression of delayed hypersensitivity by the depletion of circulating monocytes.

The intraperitoneal injection of Marcol oil was previously reported to result in the depletion of circulating monocytes. The way in which this depletion affected guinea-pigs' ability to respond to challenge with antigen was tested by challenging with tuberculin after sensitization and examining the ability to respond to DNCB. It was found that the ability to respond to tuberculin was completely abrogated by the injection of Marcol and could be restored by removing the oil from the peritoneal cavity. The ability to respond to a secondary challenge with DNCB was not affected, while the ability to become sensitized was impaired. A normal inflammatory response was seen. The role of the circulating monocyte is discussed.


Effect of protein A of Staphylococcus aureus on the binding of monomeric and polymeric IgG to Fc receptor-bearing cells.

The effect of protein A of Staphylococcus aureus (SpA) on the binding of rabbit IgG to the Fc receptors of mouse lymphocytes and macrophages was found to correlate with the aggregation of the IgG ligands. After binding anti-erythrocyte IgG complexed with SpA, the cells were able to attach to and kill erythrocyte indicator cells with a higher efficiency than lymphoid cells treated with anti-erythrocyte IgG alone. The amount of anti-peroxidase IgG which can be bound to effector cells was not changed by reaction with SpA. In contrast, the binding to cells of IgG-coated erythrocytes and of anti-peroxidase IgG complexed with peroxidase was substantially reduced by reaction with SpA. The results are compatible with the presence of two distinct Fc receptors, one for cytophilic monomeric IgG and another for polymeric (antigen-complexed) IgG.


Kinetics of humoral responsiveness and antigenic distribution in operated rats.

Wistar R/A rats were injected intravenously with 10(9) sheep red blood cells (SRBC) prior to, during or after a standard laparotomy. Stimulation of anti-SRBC antibody synthesis was already observed when the antigen was given 4 h prior to surgery and was maximal if SRBC were administered at the time of operation. The enhancing effect on the immune response lasted for 2 days after surgery. From the third post-operative day on, the injection of SRBC induced a normal humoral response. No subsequent depression was detected. Inter-organ distribution studies of 51Cr-labelled SRBC injected at various times prior, during or after the surgical procedure, showed a maximum decrease of liver uptake during operation; the depression was still present 2 h later but on the first post-operative day, no significant difference from the controls could be demonstrated. When the labelled antigen was given before surgery, organ distribution was normal. Consequently, there is no time relationship between the stimulation of antibody production and the alteration of total phagocytosis induced by surgery. Therefore, the enhanced humoral response cannot be explained only by spillover of the antigen from the liver into lymphoid organs.


Functional properties of bovine IgG1 and IgG2: interaction with complement, macrophages, neutrophils and skin.

Bovine immunoglobulin G subclass (IgG1 and IgG2) antibodies were found to fix bovine complement while only IgG1 fixed guinea-pig complement in vitro. Similar results were noted when IgG1 and IgG2 antibodies were tested by passive cutaneous anaphylaxis (PCA) in that both IgG1 and IgG2 caused PCA in bovine skin while only IgG1 mediated the reaction in rat skin. In precipitation reactions IgG1 antibodies to DNP failed to cause precipitation of DNP19-ovalbumin while IgG2 antibodies to DNP precipitated DNP19-ovalbumin. Both IgG1 and IgG2 antibodies to ovalbumin precipitated ovalbumin. Surprisingly, IgG2 antibodies to equine erythrocytes caused phagocytosis by bovine neutrophils and peripheral blood monocytes while IgG1 antibodies failed to cause either phagocytosis or adherence. Results with peripheral blood monocytes cultured for 7 days demonstrated that both IgG1 and IgG2 could mediate phagocytosis.


The responsiveness of efferent lymph cells to phytohaemagglutinin during the response of the popliteal node to dinitrophenylated bovine serum albumin.

Cells, recovered from the efferent lymphatics of the popliteal nodes of sheep during in vivo responses to dinitrophenylated bovine serum albumin (DNP-BSA), were examined for their responsiveness to phytohaemagglutinin (PHA) in vitro during the first 4--5 days of the immune response. The response was almost totally depressed when cells collected throughout the response were cultured with supra-optimal doses of the mitogen. Optimal and suboptimal doses of PHA resulted in greatly enhanced responses in cells collected in the first two periods (0--6 h; 6--12h) of the response; the presence of high doses of DNP-BSA in the culture media prevented the enhancement in the second period but had no adverse effects on the cells collected during the first 6 h. Efferent cells collected after 12 h generally showed decreased responses, this depression being maximal in cells collected 2--3 days after antigenic stimulation. Later in the response the cells again exhibited enhanced responses. The possible interpretations of these results in terms of regulation of the response are discussed.