The long-term effect of radiotherapy on the immune status of patients cured of a gynecologic malignancy. The long-term effect of radiotherapy on the immune system were evaluated in 52 patients cured for a gynecologic malignancy from 3 to 15 years. Eighty-seven percent of the patients had no depression in cell-mediated immunity as determined by delayed hypersensitivity reaction to DNCB. The absolute lymphocyte count, as well as the circulating immunoglobulins (IgA, IgG, IgM), were within the normal range. Thus, there were no significant changes in cell-mediated immunity and circulating immunoglobulins several years after radical wide-field radiotherapy.


Soldiers of Saint Agatha: some campaigns against breast cancer. Fourth Wendell G. Scott Memorial Lecture. March 10, 1976. There have been real and heartening advances in the diagnosis and treatment of breast cancer during the past 20 years. A greater proportion of women with breast cancer can now be detected before cancer has metastasized. Some patients with breast cancer can now be cured by systemic chemotherapy. Much of the problem still remains to be researched, but the control of breast cancer can be realistically predicted.


Breast adenomas.

Breast adenomas. True adenomas of the breast are cellular epithelial lesions, which may be confused with carcinoma and are difficult to classify. We have reviewed 28 adenomas of the breast and have developed specific diagnostic criteria, as well as a classification of breast adenomas based on histologic features. Electron microscopic studies confirm that adenomas have ultrastructural features similar to normal breast epithelium and are not variants of fibroadenomas. The association between lactating adenomas and pregnancy has been confirmed; however, clinicopathologic correlation does not suggest a relationship between adenomas and oral hormonal medication. Although adenomas are benign, this study includes the third reported case of an infiltrating carcinoma associated with a lactating adenoma.


Meningeal carcinomatosis.

Meningeal carcinomatosis. An anatomopathologic study of 18 cases of pure meningeal carcinomatosis is presented. In five of these cases, the brain, spinal cord, choroid plexuses, cerebral vessels, and prevertebral soft tissues, including the lumbosacral nerve plexuses and ganglia, were examined microscopically in an attmept to determine the routes of tumor spread. Our results suggest that the malignant cells reach the cerebrospinal leptomeninges via perineural, endoneural, and perivascular lymphatics and sheaths through the intervertebral and possibly cranial foramina. Involvement of the choroid plexuses appears to be secondary to, rather than the avenue for, leptomeningeal carcinomatosis, with the tumor cells reaching the choroid plexuses via the perivascular sheaths of choroidal vessels. Leptomeningeal carcinomatosis was the only manifestation of metastatic spread beyond regional lymph nodes in about 40% of all reported cases in which this information is available. This implies that radiation or other forms of local therapy to the cerebrospinal leptomeninges may provide an effective means of palliation in many of these cases.


Nonepithelial tumors of the nasal cavity, paranasal sinuses, and nasopharynx. A clinicopathologic study. VI. Fibrous tissue tumors (fibroma, fibromatosis, fibrosarcoma). In a study of 256 nonepithelial neoplasms involving the nasal cavity, paranasal sinuses, and nasopharynx, 23 lesions were classified as fibrous tissue tumors, including four cases of "fibroma", six of fibromatosis, and thirteen of fibrosarcoma. The clinical findings associated with these lesions are described, their histologic features illustrated, results of therapy presented and clinicopathologic correlations made. The "fibromas" presented a small localized nodules. None recurred after local excision. Fibromatosis, a locally aggressive tumor, does not metastasize, but may cause considerable morbidity or even death due to local infiltration which may be difficult to control surgically. Fibrosarcoma may cause death either by local infiltration or by metastasis, but has a better prognosis than most other sarcomas of this region. We recommend that a large en block resection be performed initially for fibromatosis and fibrosarcoma growing in this area, after the diagnosis has been made by biopsy. In this series, including patients who had more than one operation, recurrent tumor was seen following 10 of 12 limited local excisions performed for fibromatosis and fibrosarcoma, but in only one of 13 patients after a large bloc resection. The problems involved in histologically differentiating fibrous tissue tumors from other lesions are discussed. A patient with the rare syndrome of multicentric fibromatosis with spontaneous regression of lesions is presented.


Adenocarcinoma of the small bowel in association with regional enteritis. Four new cases. Four cases of carcinoma of the small bowel following chronic regional enteritis are presented. Search of the medical literature revealed 31 other cases reported up to the present. We must entertain the possibility that chronic regional enteritis may predispose to the development of cancer of the small bowel.


Carcinoma of the small bowel. A complication of regional enteritis. The 35th and 36th cases of adenocarcinoma arising in small bowel affected with regional enteritis are presented. The pathology of all reported cases is reviewed. An association between regional enteritis and carcinoma of the small bowel is suggested.


Subsite mapping of enzymes: collecting and processing experimental data--a case study of an amylase-malto-oligosaccharide system. Two research groups have independently developed the theory and experimental methodology for quantitatively assessing substrate monomer-subsite binding-energies for depolymerases. When the two approaches are applied to the same enzyme-substrate system they yield surprisingly divergent results. This paper outlines the application of the two approaches to an amylase-maltooligosaccharide system and points out the more important areas of disagreement. We show that by proper data-management, the conflicts between the tow laboratories are basically resolved. The complexities of the subsite model demand extensive data-gathering and exacting data-processing and verification that the computed model-parameters can faithfully reproduce the experimental data.


Synthesis of beta-nitrophenyl 2-acetamido-2-deoxy-3-O-beta-D-galactopyranosyl-D-glactopyranosides. Reaction of 2,3,4,6-tetra-O-acetyl-alpha-D-galactopyranosyl bromide with p-nitrophenyl 2-acetamido-2-deoxy-4,6-O-(p-methoxybenzylidene)-alpha-D-galactopyranoside under the usual conditions, followed by removal of the p-methoxybenzylidene group and O-deacylation, gave crystalline p-nitrophenyl 2-acetamido-2-deoxy-3-O-beta-D-galactopyranosyl-alpha-D-galactopyranoside. The synthesis of p-nitrophenyl 2-acetamido-2-deoxy-3-O-beta-D-galactopyranosyl-beta-D-galactopyranoside has also been accomplished by a similar reaction-sequence.


Alternation in mitral valve and septal movements in a patient with pulsus alternans. In a patient with recent aortic valve replacement, echocardiography, phonocardiography, and external pulse recordings were employed to evaluate a prolonged period of pulsus alternans. Alternation in left ventricular end-diastolic and end-systolic diameters, systolic diameter shortening, diastolic ventricular filling, systolic ejection time and diastolic period (A2Q) were seen. In addition, two features were noted which have not been previously reported: (1) Alternation in the timing of mitral valve motion, such that prior to the weak beats, closure of the mitral valve did not occur until 40-70 msec. after the onset of the Q-wave on the ECG. Before the strong beats, however, mitral closure coincided approximately with the Q-wave. (2) Alternation of the movement of the interventricular septum with abnormal or paradoxical movement during weak beats and normal movement during the strong beats. The first observation lends credence to the theory that stretch-tension relationships are important in the maintenance of pulsus alternans in some patients. The paradoxical septal motion during the weak beats may be due either to a discordant right and left ventricular alternans or alternate contractile defection of the septum.


Pulmonary vascular complications in narcotic addicts. Perfusion deficits on lung scanning in narcotic addicts are frequently demonstrable. To determine the angiographic and hemodynamic correlates of these deficits, right heart catheterization with selective pulmonary angiography was carried out in 7 addicts with abnormal lung scans, and one with normal scan. Four had angiographic abnormalities corresponding to the regions of perfusion defects on lung scan. These consisted of large vessel cut-offs, intraluminal filling defects and/or considerably reduced vascularity. Three subjects had normal angiograms in the presence of abnormal perfusion scans. One subject had normal angiogram and scan. All eight patients had normal pulmonary artery pressures and calculated pulmonary vascular resistances. This study demonstrates that (1) angiographic abnormalities in larger pulmonary arteries may be found in narcotic addicts with perfusion defects on lung scanning, and (2) these findings may be present without significant hemodynamic alterations.


Interaction of rabbit sperm and egg. Examination of the same cryofractured specimens with scanning and transmission electron microscopy. Rabbit ova fertilized in vitro were prepared for scanning electron microscopy by ethanol-cryofracturing and critical-point drying methods and were also embedded and sectioned for transmission electron microscopy. Study of a region of interaction between sperm and zona pellucida with scanning electron microscopy reveals the latter to be composed of a complex network of fibers interspersed with numerous pores. Transmission electron microscopy of the same region reveals a "typical" homogeneous composition of the zona pellucida. Ultrastructural observations of thin sections passing through the region of sperm-egg interactions or through other regions of the ovum or its investments reveals very little methodological distortion of the various intracellular organelles or matrix. Application of the procedures described provides not only an elucidation of surface detail but also reveals intracellular cytoplasmic information about the same specimen during in vitro fertilization.


Differential survival of isolated portions of crayfish axons. Electron microscopic studies show that transplanted segments of sensory axons of varying lengths degenerate within 7-14 days whereas transplanted segments of crustacean motor axons survive morphologically intact for 20-30 days. The middle portion of an isolated motor axon segment degenerates less rapidly than portions of the same axon located nearer the periphery or nearer the ventral nerve cord. One week after transplantation, glial cells appear to phagocytize sensory axons whereas glial cells around motor axons appear to hypertrophy and to have more rough endoplasmic reticulum. After three weeks, motor axons also appear to be phagocytized by glial cells. These data suggest that the glia surrounding isolated motor axons can change from a supportive to a destructive function, whereas glial cells surrounding severed sensory axons primarily have a destructive function. These and other data also indicate that crustacean motor axons receive significant trophic inputs from their own perikaryon, from post-synaptic contacts, and from adjacent glial cells. The possibility that adjacent healthy cells may supply metabolically deficient cells with needed substances could be a significant adaptive advantage for the evolution of multicellular organisms.


Immunocytochemical localization of nerve growth factor (NGF) in the submandibular gland of adult mice by light and electron microscopy. Nerve growth factor (NGF) was localized in the submandibular gland of adult male mice by a direct immunocytochemical method using highly purified antibodies against NGF coupled to horseradish peroxidase. In light microscopic sections the reaction product was entirely confined to the cells of the secretory tubules. The acinar part of the gland was free of reaction product. This finding was confirmed by electron microscopy. Within the cells NGF was localized exclusively in the apical secretory granules. No reaction was observed in the rough endoplasmic reticulum, the Golgi region or in the granules of the basal part of the cells. This observation favours the assumption that NGF is derived from a precursor molecule and that the precursor is transformed into immunologically active NGF within the secretory granules during their transport from the basal to the apical part of the tubular cells. Stimulation of the submandibular gland with carbachol (2 mg/kg) led to a massive release of the content of the secretory granules, including NGF, into the salivary duct.


The early development of the tail and the transformation of the shape of the nucleus of the spermatid of the domestic fowl, Gallus gallus. The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl. In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum. The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.


The glial cells of the cerebral ganglia of Helix pomatia L. (Gastropoda, Pulmonata). II. Uptake of ferritin and 3H-glutamate. 1. With Helix pomatia intracerebral injections of ferritin were carried out (maximal incubation time: 45 min). First, the marker spreads with time via the enxtracellular space throughout the cerebral ganglia and, secondly, is transported out of the ganglia. Electron microscopical studies showed that all glial cell types take up great amounts of ferritin by endocytosis. The plasmatic glial cells at the periphery incorporate more of the marker than the filamentous glial cells in the centre. No uptake of ferritin by neurons or axons was observed. In vitro studies proved that ferritin can penetrate from the connective tissue capsule into the ganglia only after disruption of the neural lamella and damaging of the peripheral glial processes. 2. 3H-glutamate, a putative transmitter of the CNS of Helix pomatia, was injected into the hemocoel of active snails (incubation times: 15 min, 1h, 6h, 3d). Light microscopical evaluation of radioautographs showed that great quantities of the tracer penetrate into the ganglia. The bulk of it is taken up by glial cells, whereas the neurons exhibit only small amounts of the tracer. The studies with ferritin as well as those with 3H-glutamate indicate that the glial cells of the cerebral ganglia of Helix pomatia act as a "hemolymph-neuron barrier". A dominant role of the plasmatic glial cells according to these processes is discussed.


Ultrastructure of the salivary bladder of the nine-banded armadillo. The nine-banded armadillo possesses a salivary bladder which is a dilated portion of the main duct of the submandibular gland at its origin. The wall of the bladder is composed of an epithelium, a submucosa and a thick coat of skeletal muscle. The ultrastructure of the epithelium reveals that it is complex and consists of three cell types: 1) principal cells, 2) light cells, and 3) basal cells. The general organization of the epithelium suggests that it is a transporting type of epithelium such as that found in the amphibian and reptilian and reptilian urinary bladders and the mammalian gall bladder. The submucosa is composed primarily of densely-packed collagen fibers. The skeletal muscle is very vascular and richly innervated.


Innervation of the synovial membrane of the cats joint capsule. Results of investigations on the occurrence of nerve fibres and endings in the synovial membrane of the knee and elbow joint in the cat are reported. The stratum synoviale contains only autonomic fibres, running in the adventitia of arteries. Free nerve endings are lacking in the stratum synoviale. Simple Pacinian corpuscles with an inner core are occasionally observed in the border zone between the stratum synoviale and fibrosum. The ultrastructure of these endorgans resembles that of Pacinian corpuscles in the hairless and hairy skin of the cat.


Morphology, morphometry and electron microscopy of HeLa cells infected with bovine Mycoplasma. The host-parasite relationship of HeLa M cells artificially infected with a bovine species of Mycoplasma was studied by light microscopy, transmission electron microscopy and scanning electron microscopy. The use of morphometry to quantitate some of the findings was explored. The parasites were seen in locations extracellular to the cell surface. The detection of small numbers of organisms by light microscopy was well demonstrated by use of the flurescent antibody technique. Scanning electron microscopy proved to be an excellent method for revealing the surface details of cell-parasite morphology. Ultra-thin sections showed that the parasites are aligned mostly parallel to the plasma membrane of the host cell but separated by a gap of 10nm. Morphometry indicated an average of 69 organisms per cell surface occupying 1.7% of the surface area. An increase of 26% in diameter of the HeLa cells, possibly as a result of infection, was observed.


The hypothalamo-hypophysial system in Acipenseridae. V. Ecological-histophysiological analysis of the neurohypophysis of the female sturgeon Acipenser güldenstädti Brandt during up-stream migration and after spawning. Neurohypophyses from sexually mature female sturgeons, Acipenser guldenstadti Brandt, killed before, soon after spawning, and during down-stream migration were studied light and electron microscopically. Ovaries were examined only under a light microscope. A large amount of neurosecretory material (3.5-4.5 arbitrary units) is found in the neurohypophysial "roots" during up-stream migration to spawning grounds. Neurosecretory fibres and their terminals are replete with elementary neurosecretory granules. Side by side with the latter some terminals contain single residual granules and few "synaptic" vesicles. Soon after spawning the amount of neurosecretory material decreases markedly in most individuals (content up to 1-3 units), and elementary granules are few in the fibres and their terminals. The number of residual granules increases, and "synaptic" vesicles are especially numerous. Granulated, disintegrating neurosecretory granules and granule-shadows occur in the preterminal parts of the fibres. The number of pituicytes increases becuase some "light" tanycytes seem to migrate from the ependymal into the subependymal layer. The restoration of neurosecretory material (reaching again up to 3.5-4.5 units) occurs within a month after spawning. The number of elementary granules increases in fibres and terminals while that of residual granules and "synaptic" vesicles decreases. These changes in the neurohypophysis of females after spawning are due to discharge of great amounts of peptide neurohorsmones into the general circulation after spawning. Spawning is regarded as a "physiological" stress for these females.


The hypothalamo-hypophysial system in acipenseridae. VI. The proximal neurosecretory contact region. The proximal neurosecretory contact region (PCR) of Acipenseridae, a homologue of the tetrapod median eminence, has been studied by light, fluorescence and electron microscopy. It occupies the rostral and chiefly the ventral surfaces of the hypothalamic tuber cinereum. PAF-positive fibres occur in the zone of the preoptico-hypophysial tract but their terminal enlargements are concentrated mainly in the external zone. They make contact with the primary portal capillaries situated in the pia mater. Monoaminergic fibres and terminals with an intense green fluorescence are localized in the same regions. The fibres of some bipolar monoaminergic neurons of the PCR make contact both with the third ventricle and the primary portal capillaries. Three types of granule-containing neurosecretory fibres and terminals have been recognized in the PCR. Fibres of types A1 (d = 120-300 nm) and A2 (D = 100-170 nm) are peptidergic PAR-positive, although some fibres, including some of type A1, belong possibly to PAF-negative type. Monoaminergic type B fibres have granules 80-100 nm in diameter. Neurosecretory terminals and vascular "endfeet" of tanycytes make contact with the 70 nm thick outer basement membrane of the primary portal capillaries. Several laminae of thin horizontally oriented tanycyte processes form a boundary between the external zone and the preoptico-hypophysial tract. Few neuroglial cells with pale cytoplasm, numerous lysosomes and lipofuscin granules are seen in this region. It is hypothesized that, as in other vertebrates both peptide hypophysiotropic neurohormones and monoamines are discarged from the PCR into the portal circulation and affect the activity of the glandular cells of the pars distalis.


The large pigment cell of the compound eye of the house fly Musca domestica. Fine structure and cytoarchitectural associations. The fine structure and cellular associations of the large pigment cells (LPC's) of the compound eye of the house fly were studied with high voltage and conventional electron microscopy. Depending on the sector of the compound eye, the facets are either rectangular or hexagonal. The underside of each facet has indentations exactly aligned with those on top into which inserts an angulated sleeve of LPC's. Under the rectangular lens facet 6 or 8 small compact (in cross section) LPC's join four elongate LPC's. Clusters of compact cells alternate in this ring with elongate ones. Compact cells compress together and become quadrangular (in cross section) several microns below their insertion into the lens and form "building block" corners while elongate cells form "side rails" for the rectangular type of distal pseudocone enclosure. Beneath hexagonal facets all LPC's are rather elongate with out corner cells. In both facet types LPC's enclose the pseudocone for a longitudinal distance of 4 mum and then are displaced as bordering cells by a sleeve of two corneal pigment cells (CPC's), each of which encloses half of the proximal pseudocone. For the following 6 mum of longitudinal distance these concentric sleeves of CPC's and LPC's form a double layer around the pseudocone. At about 10 mum below lens base the two sleeves separate; LPC's become attenuated and extend cable-like to the basement membrane and CPC's enclose the proximal pseudocone, Semper cells and distal retinula. The junction between lens and LPC's has critical structural value in that (1) this is the sole anchorage to the lens by the lengthy remainder of the ommatidium, and (2) LPC's enclose the semiliquid pseudocone in the most distal portion of the pseudocone. In addition to vertical support, the LPC's send out numerous lateral processes that make structural contact among themselves, with the corneal pigment cells and the photoreceptor cells. The structural features of this array are discussed relative to possible physiological roles.


Multiple rough endoplasmic cisternae in "C" cells. Multiple rough endoplasmic cisternae were found in "C" cells of the adult rat, at interphase. They are considered to be normal constituents of "C" cells. Their morphological relation to rough endoplasmic reticulum and their close proximity to mitochondria, Golgi dictyosomes and secretory granules suggest that they may have a role in the secretory activity of this endocrine cells.


Fine structure of baroreceptor terminals in the carotid sinus of guinea pigs and mice. A light and electron microscopic study was undertaken on the baroreceptor axon terminals in the carotid sinus of guinea pigs and mice, using serial semithin and thin sections. Together with their enveloping Schwann cells, numerous lanceolate axon terminals are organized into a well-defined discoid end organ, referred to as the 'baroreceptor unit'. Baroreceptor units measure 100 to 150 mum in diameter and are arranged in a hexagonal pattern. These end organs represent free branched lanceolate mechanoreceptors of complex type (Andres and von Düring, 1973) which belong to the main group of stretch receptors. In the guinea pig the lanceolate terminals enter the media and approach the innermost layers near the intima. In the mouse the terminals are seen to spread in the adventitia and along the medio-adventitial border. Only a few of them penetrate the external elastic layer. Species differences concerning the localization and extent of these visceral mechanoreceptors are discussed, as well as the modified architecture of the sinus wall in the receptor area ('elastic segment'). Lanceolate terminals form beaded varicosities which are equipped with finger-like or lamellar exoplasmic protrusions. These projections contain a well-differentiated receptor matrix. They are attached to collagen and elastic fibers. The varicosities include densely packed mitochondria, neurotubules, profiles of axoplasmic reticulum, clear and granular vesicles, and striking accumulations of glycogen particles, lamellated bodies and lysosomes. Four types of varicosities are discerned according to their main axoplasmic components. Various types of these varicosities occur within an individual lanceolate terminal.


Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. When their growth is arrested in culture, susceptible 3T3 fibroblasts differentiate into adipose cells. Different clones form adipose cells with different frequency, depending upon the proportion of susceptible cells they contain. In cultures grown from small inocula, the fat cells appear in clusters formed by colonies of susceptible cells. Study of these clusters indicates the infrequent occurrence of cellular transitions from insusceptible to susceptible state. Beginning with a clone converting to adipose cells with a vary low frequency, it has been possible, by serial selection, to generate subclones which convert with a high frequency. This evolution is due to spontaneous heritable changes affecting susceptibility to the adipose conversion. Presumably, they involve the control of triglyceride synthesis. Early stages of the adipose conversion may be recognized in stained cultures. When triglyceride first begins to accumulate, the highly extended and flattened processes of the cells are probably similar to those of nonfatty cells in the same cultures. As the adipose conversion proceeds, the processes thicken and retract; the cells eventually acquire the rounded shape of the more mature adipose cells.


The regeneration and division of mouse L-cell karyoplasts. A technique for efficient cytochalasin-induced enucleation was used to prepare "karyoplasts"--nuclei surrounded by a thin shell of cytoplasm and an outer cell membrane. Methods for estimating the quantity of cytoplasm remaining in karyoplasts indicated that they contained less than 10% of the amount found in whole cells. Procedures for separating karyoplasts from contaminating cytoplasmic fragments and whole cells are also described. Freshly prepared L-cell karyoplasts were unable to adhere to and spread upon a surface. However, after incubation for several days, about 30% of the karyoplasts regained these abilities to some degree. A portion of the regenerating karyoplasts were then observed to divide. These events were confirmed and recorded by time-lapse cinematography. In addition, by culturing karyoplasts under appropriate conditions, clones were isolated.


Direct induction by estradiol on vitellogenin synthesis in organ cultures of male Xenopus laevis liver. Organ cultures of liver from untreated male Xenopus respond to 17 beta-estradiol in the culture medium by synthesizing and secreting the yolk protein precursor vitellogenin. Vitellogenin synthesis, as a primary response, is first detectable on the fourth day of culture, and comprises up to 12% of the protein synthesized on the eighth day. Estradiol is required during the lag period of 4 days. Tissue from male Xenopus injected with 1 mg of estradiol 30 days before the start of culture responds more rapidly and to a greater extent to estradiol in the medium than tissue from uninjected males. During such a secondary response, vitellogenin is first detectable in the medium on the second day of culture, and becomes up to 24% of the protein synthesized on day 6. The rate of amino acid incorporation into total protein also increases in response to estradiol, but the rate of synthesis of albumin decreases rapidly in culture whether or not estradiol is present, in both the primary and secondary responses. A maximal response is seen with 10(-8) M estradiol. Progesterone, testosterone, dexamethasone, and insulin neither induce vitellogenin synthesis in culture nor modify the response to estradiol. DNA synthesis inhibitors do not prevent the response to estradiol in vitro, suggesting that cell division is not required for the initial response leading to vitellogenin synthesis.


Location and identification of the genes for adenovirus type 2 early polypeptides. Virus-specific RNA was prepared from cells early after adenovirus type 2 infection and fractionated by hybridization to specific fragments of viral DNA. The viral mRNA was used to program cell-free protein synthesis, and the products were analyzed by electrophoresis. The genes for the early polypeptides of apparent molecular weight 44,000, 15,000, 72,000, 15,500, 19,000, and 11,000 daltons were located, respectively, between positions 0-4.1, 4.1-16.7, 58.5-70.7, 75.9-83.4, 89.7-98.6, and 89.7-98.6 of the conventional adenovirus DNA map. The polypeptide of molecular weight 72,000 daltons was shown to be the single-stranded DNA-binding protein described by others. RNAs from three different adeno-transformed cell lines each program the synthesis in vitro of predominantly the 15K polypeptide, as well as variable amounts of the polypeptide of molecular weight 44,000 daltons. The genes for these two polypeptides are located in the portion of DNA known to be required for transformation of rodent cells by adenovirus.


Preliminary characterization of the temperature-sensitive defect in DNA replication in a mutant mouse L cell. Temperature-sensitive ts A1S9 mouse L cells synthesize DNA apparently normally for 6-8 hr upon incubation at 38.5 degrees C. Thereafter, these cells are able to perform limited polydeoxyribonucleotide chain synthesis at the high temperature, but are unable to convert newly replicated small single-strand segments of DNA (of the order of molecular weight 10(6) daltons) to large molecular weight chromosomal DNA. Data obtained are compatible with a model which suggests that ts A1S9 cells are able to carry out most individual reactions of DNA synthesis at the high temperature, but are temperature-sensitive in a protein which participates in the joining of small DNA segments to make chromosomal DNA strands. When cells are reincubated at a permissive temperature, after the temperature-sensitive lesion has been established, they recover the latter capability several hours before they are able once again to synthesize DNA at normal rates.


Different lifetimes of reticulocyte messenger RNA. About 85-90% of cytoplasmic protein synthesized by young reticulocytes is globin, and about 10% is a polypeptide (I) of molecular weight 64,000 daltons. Maturation of reticulocytes is accompanied by selective reduction in the synthesis of polypeptide I, relative to globin; mature reticulocytes synthesize globin at a high level but make no detectable polypeptide I. Studies in which RNA from young and old reticulocytes was translated in a wheat germ cell-free extract showed that reduction in synthesis of polypeptide I was correlated with a reduction in the amount of translatable mRNA for this protein. Differential destruction of mRNA thus is an important factor in determining the types of proteins made during the final stages of erythropoiesis.


Modifications of mammalian cell surfaces induced by sugars: scanning electron microscopy. Substitution of galactose, xylose, or mannose for glucose in the growth medium of L cells or the addition of equal concentrations of the alternate sugars to glucose-containing medium results in marked morphologic changes. The changes are revealed by conventional staining for light microscopy and by scanning electron microscopy. L cells grow indefinitely on combinations of equal concentrations of glucose and galactose, xylose, or mannose. There appear to be no significant differences in growth rate on glucose compared to the combinations of sugars cited. Cells subcultured from glucose to the combinations while undergoing rapid multiplication show marked morphologic changes by light and scanning electron microscopy within 36 hr. Of particular note are the loss of microvilli; flattening of the cells; assumption of polygonal shape; prominence of nuclei and nucleoli; and changes in the structure and distribution of filopodia. Virtually all cells in the population exhibit the changes noted.


Tracer studies of cholesterol degradation induced by ionized gases. Multiple molecular layers of cholesterol-4-C14 were bombarded in an ultra high vacuum system with various excited molecular, ionic and atomic species derived from oxygen, methane and carbon monoxide. The degradation products were analyzed by thin layer chromatography followed by autoradiography and the decomposition patterns were compared with those obtained from the autoxidation of cholesterol. Bombardment with excited and ionized species derived from both molecular oxygen and carbon monoxide leads to a number of known polar cholesterol oxidation products, similar to those formed during the natural oxidation of cholesterol. In contrast, species derived from methane lead only to the formation of products less polar than cholesterol with none of the established autoxidation products at detectable levels.


Molecular aggregation of the slow reacting hemolytic lysolecithin analog 1-octadecyl-2-benzyl-glycero-3-phosphorylcholine in aqueous solution. An attempt has been made to relate the retarded adsoprtion to red cells of the slow reacting hemolytic phosphatide Rac. 1-octadecyl-2-benzyl-glycero-3-phosphorylcholine (benzyl-lysolecithin) to its aggregation status in aqueous solution. Light scattering measurements indicate a critical micelle concentration at 37 degrees of less than 2 X 10(-6) M. The micellar weight as determined by angle dependent light scattering is 6 X 10(7) with about 97 000 molecules per micells. The aggregates, which according to electron-microscopic observations are more similar to lecithin-liposomes than to usual lysolecithin-micelles, undergo a phase transition at 14 degrees from a tightly packed liquid-crystalline state to the more loose structure of a gel phase with increased mobility of the aliphatic chains. The enthalpy of transition is 4.2 kcal/mole. These changes of the micellar structure are reflected in the binding kinetics of benzyl-lysolecithin to red cells in that the binding rate is rather constant below, but strongly increasing above the transition temperature. It is concluded that the unusual micellar structure is responsible for the retarded adsorption of this lysolecithin analog to red cells and that the rate of adsorption is probably determined by the rate of escape of single lysophosphatide molecules from the micelles.


Synthesis and properties of 35S, 14C and 3H labeled S-alkyl glycerol ethers and derivatives. Radioactive S-alkyl glycerol ethers have been synthesized with 35S, 14C and 3H labels as well as 3H/35S double labels. The synthesized compounds were converted to various derivatives which can serve to characterize the S-alkyl glycerol ethers. These included the isopropylidene derivative, oxidation with periodate to the aldehyde followed by reduction with LiAlH4 to the alcohol, and reaction of the alcohol with acetic anhydride to form the acetate derivative. Chemical analysis, IR, NMR, zonal TLC profile scans and GLC showed all the products to be greater than 99% pure. The GLC behavior of the aldehyde and acetate derivatives of both S-alkyl glycerol ethers and O-alkyl glycerol ethers on EGSS-X was compared.


Profile: the Pre-School Playgroups Association--playgroups in the community. From 12-19 June, the Pre-School Playgroups Association is holding National Playgroup Week. This week is being celebrated by the PPA's 10,000 members in England and Wales and its sister organizations in Scotland and Ireland. Playgroups came into being 15 years ago and their numbers have mushroomed in the last few years. In this article we look at the contribution a playgroup can make to the community.


X-Prolyl dipeptidyl-aminopeptidase activity, with X-proline p-nitroanilides as substrates, in normal and pathological human sera. X-Prolyl dipeptidyl-aminopeptidase (no EC no. assigned) activity in normal and pathological human sera was assayed with several newly synthesized X-proline p-nitroanilides as chromogenic substrates. Normal values for 88 healthy subjects (15 to 81 years old), with glycylproline p-nitroanilide as substrate at pH 8.7, were 54.9 +/- 1.5 (SE) (range, 25.7 - 96.0) mumol/min per liter of serum at 37 degrees C. The results suggest that the enzyme activities with all X-proline p-nitroanilides were increased in patients with hepatitis and decreased in patients with gastric cancer. On Sephadex G-200 column chromatography, normal human sera showed a single peak of enzyme activity with glycylproline p-nitroanilide as the substrate, which coincided with the peak with glycylproline beta-naphthylamide but was different from the peaks with leucine beta-naphthylamide. Sera from patients with hepatitis or liver cirrhosis showed an increase in the normal peak and the appearance of another new peak with glycylproline p-nitroanilide as substrate.


Captured for consideration--using videotape as an aid to the treatment of the disturbed child. The use of videotape as a clinical tool to monitor work with children is discussed with reference to a group of three emotionally disturbed children seen weekly in a paediatric assessment and treatment centre. Six particular attributes of videotape are distinguished and illustrated from clinical material: video is a vehicle for discussion; video permits detailed observation; video see things that humans do not; video has a distancing effect; video shares information and video trains in observation and handling techniques. It is suggested that the experience derived from using videotape intensively over a short period can be applied to advantage in other clinical situations.


Immunofixation electrophoretic techniques applied to identification of proteins in serum and cerebrospinal fluid. We describe the application of immunofixation staining of agarose-gel electrophoretograms in areas where its use in the clinical laboratory is appropriate. Immunofixation electrophoresis consists of an electrophoretic phase followed by a fixation phase in which antiserum is used to precipitate the protein. As long as the antibody is in slight excess or near equivalency, the antigen/antibody complex remains insoluble. The reaction can be detected by visual inspection in indirect light, by protein staining, or by use of antibodies labeled with fluorescein, enzyme, or isotope. In the method described here we primarily have used protein staining (Coomassie Blue) to accentuate the proteins fixed by antisera. All unreacted proteins are removed by pressing with filter paper and saline washing. In the clinical laboratory, this method expedites immunochemical evaluation of samples and may also supplement immunoelectrophoresis. It has been applied successfully in identifying small obscure monoclonal proteins in the serum and cerebrospinal fluid of patients with multiple sclerosis, subacute sclerosing panencephalitis, biclonal gammopathies, serum monoclonal light chains, and mobility shifts of certain proteins, particularly of the complement series. Immunofixation demonstrates that the protein bands present in spinal fluid from multiple sclerosis and subacute sclerosing panencephalitis patients are of the IgG class of immunoglobulins; and non-IgG protein, such as beta and gamma trace proteins, are not detected. We also comment on reverse immunofixation with labeled antigen as a branch of the procedure that allows detection of function of the immunoglobulins separated by electrophoresis.


Catabolism of plasma enzymes, as studied with 125I-labeled lactate dehydrogenase-1 in the rabbit. Circulating enzymes may be inactivated in the plasma and the inactive breakdown products may be hydrolyzed in the lumen of the small intestine. Evidence for this mechanism was based upon previous studies with 125I-labeled lactate dehydrogenase-5, and here similar studies with radioiodinated lactate dehydrogenase-1 are reported, to determine whether this isoenzyme is similarly catabolized. The pure rabbit enzyme was labeled with 125I by use of lactoperoxidase and hydrogen peroxide (the labeled enzyme had 80-85% of the original catalytic activity). After its intravenous injection into rabbits, plasma enzyme activity and radioactivity disappeared during the first 4 h at similar fast rates, apparently because of distribution of the injected enzyme throughout the extracellular fluid. During a second phase (30-h), catalytic activity disappeared significantly faster than radioactivity, suggesting inactivation of the enzyme in either the plasma or a compartment in close proximity to it, or both. Enzyme activity then remained constant while plasma radioactivity continued to decrease at a slower, exponential rate, apparently owing to removal of breakdown products. In no case did tissue radioactivity, studied 6 h after injection, approach that of plasma. We therefore conclude that removal of the enzyme protein or its breakdown products is a passive process. Appreciable radioactivity was detected in the intestinal contents, a finding which suggests that removal via the small intestine is an important route for the removal of inactivated enzyme products from the circulation. Less than 3% of the injected radioactivity appeared in the feces during the first three days; urinary excretion accounted for about 67% during the same period, about 60% of which consisted of radio-iodinated amino-acids, the remainder of iodide. Free mono- and di-iodotyrosines were among the products excreted. These appear to originate from absorption of the products of further breakdown of the enzyme molecule in the intestine.


Rates of disappearance from plasma of enzymes labeled by coupling with a radioactive lodo-ester. Lactate dehydrogenase-5 and creatine kinase from rabbit muscle were labeled by coupling with N-hydroxysuccinimidyl 3-(4'-hydroxy-[3',5'-125I]diiodophenyl)propionate. After purification, the analytical recovery of catalytically-active labeled enzyme averaged 90% for lactate dehydrogenase, 81% for creatine kinase. The labeled enzymes were injected intravenously into rabbits and disappearance from plasma of catalytic activity and radioactivity was measured. The disappearance curves for lactate dehydrogenase-5 differed considerably from those observed with the enzyme labeled by direct iodination. The discrepancy was due to rapid hydrolysis in vivo of the labeled amide-enzyme linkage, because about 50% of the injected radioactivity appeared in the urine as 125I-labeled 3-(4'-hydroxy-3',5'-diiodophenyl)propionic acid within 4-8 h of injection. Similar outputs were observed after administration of this acid to rabbits. The free acid was also detected in the urines of rabbits within 4-8 h of the intravenous injection of creatine kinase labeled similarly. We conclude that this method of labeling is unsuitable for preparing radioactive enzymes for study of their catabolism.


Urinary organic acids in man. I. Normal patterns. We studied qualitative pattern of urinary acidic metabolites excreted by normal persons. The results provide a basis on which to compare results for patients with potentially abnormal organic acidurias. A series of urinary polyhydroxy (aldonic and deoxyaldonic) acids has been identified. Most of these compounds have not been previously reported in human urine, except in connection with the present work, and are additional to the previously recognized urinary organic acids, which were also observed. Possible metabolic origins of some of the acids are briefly discussed.


A study of some factors concerned in the schooling of visually handicapped children. Sixty children integrated into the fully-sighted school system and 34 children attending partially-sighted schools were assessed. All were using low vision aids. It was found that there was little difference in acuity, parental or school support, but the integrated children tended to be more intelligent and came from a higher socioeconomic group. More children could avoid special schools if given appropriate services at an early age and assessment should be by a multidisciplinary team.


Urinary organic acids in man. II. Effects of individual variation and diet on the urinary excretion of acidic metabolites. We have assessed individual variations in urinary acidic metabolite excretion and the effects of extreme alterations in dietary composition on these metabolites in selected normal persons who we considered representative of the general ambulant normal population. Extreme dietary alterations produced relatively small changes in the patterns or amounts of metabolite excretion, but large individual within-subject variations were observed. Our results indicate that variation in the ranges of excretion for the normal population mainly depend on individual metabolic variations rather than on dietary factors, and provide a basis for the assessment of the normal ranges determined from population surveys. Our results are discussed in relation to previous studies on the variability of urinary acidic metabolite excretion in man.


Urinary organic acids in man. III. Quantitative ranges and patterns of excretion in a normal population. We studied the urinary excretion of organic acids by normal human subjects in an ambulant normal population, chosen without conscious bias. Quantitative ranges and frequency distribution patterns of excretion are reported, as a basis on which to compare results obtained for patients whose clinical condition suggests that their excretion values may be abnormal. The quantitative valuse we observed enable the relative significance of different urinary anionic metabolites to be assessed, including the recently identified aldonic and deoxyaldonic acids. The frequency distribution patterns observed are discussed briefly.


Addition of triglyceride to serum for use in quality control and reference. We describe a simple method in which a water-soluble mixture of triocatanoin and a surfactant, "Triton X-114," are used in preparing solutions of triglycerides (triacylglycerols) in either human serum, solutions of albumin, or water. Analytical recovery added triglyceride was quantitative by two methods. The addition did not affect results of analyses for 18 other commonly measured constitutents of serum. When the triglyceride was added to either lipid-depleted human serum or bovine serum albumin solution and lyophilized, subsequent solutions were clear. The triglyceride/protein preparation was stable in lyophilized form for a year and in reconstituted serum for five days at 5 degrees C. Aquenous solutions appear to be stable indefinitely at room temperature.


Rapid-flow enthalpimetric determination of urea in serum, with use of an immobilized urease reactor. We used a differential thermal detector in conjunction with an immobilized urease reactor to determine urea in serum. Samples (120 mul) are introduced into a flow stream and passed through an "adiabatic" column, which is packed with enough insolubilized urease to completely convert urea to ammonia and carbon dioxide. Measured temperature changes are directly proportional to the serum urea concentration. Urea in the presence of protein, bilirubin, and hemoglobin can thus be rapidly, simply, and inexpensively measured. Results correlate well with those obtained by the manual diacetyl monoxime and urease/indophenol methods.


Propoxyphene and norpropoxyphene concentrations in blood and tissues in cases of fatal overdose. Propoxyphene and its major metabolite, norpropoxyphene, have been quantitated in tissue specimens obtained from autopsies of people suspected of dying from propoxyphene overdosage. Gas-chromatographic determination of both propoxyphene and norpropoxyphene is essential because the blood concentration of the parent drug should be about 1.0 mg/liter or greater to attribute a death to the drug. The metabolite concentration in blood may help to establish when the drug was ingested. Concentrations in the blood after high oral therapeutic doses are about 0.3 mg of propoxyphene per liter, and norpropoxyphene concentrations may be as high as 3 mg/liter. Methods of determining propoxyphene are discussed.


A multi-axial classification for the education of autistic children. Following the WHO tri-axial classification of mental disorders, which set out possibilities for differential classification and diagnosis among the syndromes, the present article suggests complementary axes upon which educational therapy can be based in the psychoses of autism and allied disorders. They are offered as a basis of common ground for discussion in multidisciplinary teams.


Micromethod for analysis for chloride in physiological fluids. A micromethod for the analysis for chloride, based on the chemical precipitation of silver chloride by radial diffusion through agar gel containing silver nitrate, is described. The method is simple to run, requires little or no instrumentation, and requires only 10 mul of sample. Results by coulometric titration (Buchler Cotlove Chloridometer) correlated well for serum (r = 0.961), urine (r = 0.997), cerebrospinal fluid (r = 0.991), and sweat (r = 0.998). Other halide ions or protein do not interfere. Precision studies gave a within-day reproducibility (CV) of 1.3% and a day-to-day variability of 2.1% for a serum sample averaging 115 mmol/liter.


Some theory of reference values. II. Comparison of some statistical models of intraindividual variation in blood constituents. Three models of intraindividual variation are reviewed, and statistical methods for distinguishing among them are discussed. Application of these methods to short series of observations from healthy individuals indicates that, in the large majority of cases, a strictly homeostatic model is appropriate for such constituents as serum calcium and magnesium. In less closely controlled variables, e.g., serum cholesterol and uric acid, a nonstationary, "rndom walk" model appears moresuitable in most cases. A more general autoregressive model, which includes the other models as extreme cases, could be used to describe all degrees of homeostatic control. This model is more complex, however, and requires at least 10 observations to yield estimates of acceptable precision. Moreover, it is sensitive to fluctuations in within-batch analytical variance. When biological variance is small relative to analytical variance, all three models yield essentially the same predicated values. To illustrate their use, these models have been applied to four short individual series of cholesterol observations showing increasing amounts of intrapersonal variation over long periods of time. I suggest that when less than 10 observations over time are available, the strictly homeostatic model and the nonstationary model be used to derive a "critical range" for assessing future changes. When longer series are available, the more general model might replace the other two for this purpose, if analytical variation has remained reasonably stable (within +/- 20% of its average value) during the period of observation. Much more experience with the use of all three models in health monitoring programs would be highly desirable.


Improved method for determining serum protein concentrations in the far ultraviolet. We examined the feasibility of measuring absorbance in the far ultraviolet for serum protein determination with a medium-priced, double-beam spectrophotometer. The method has been simplified by using an automatic dilutor to dilute the serum, and a flow-through cell and recorder in conjunction with the spectrophotometer. Because it is only necessary to dilute the serum and determine its absorbance, the procedure is quite rapid. As compared to the biuret reaction, interferences from hemolysis have been about halved. Interferences owing to light scattering by turbid sera have also been appreciably decreased by use of an anionic detegent (sodium dodecyl sulfate) as a serum diluent. Comparisons of the present method to the biuret and Kjeldahl methods indicate that it is relatively accurate, and is practical for routine use.


Analysis of whole blood by neutron activation: a search for a biochemical indicator of neoplasia. We determined a total of 13 elements in the whole blood of cancer patients and controls by using instrumental neutron activation analysis. The samples were spotted as whole blood on small filter paper disks, allowed to dry at room temperature, and sealed between two polyethylene disks. They were then irradiated and counted. Eleven elements were determined by this method. Sodium and chlorine were measured by irradiating whole blood in polyethylene vials. Statistical analysis of the data predicts that, with a very hgih probability, cobalt and iron are lower, adn sodium, tantalum, lanthanum and chlorine are higher in the cancer patients than in the normal persons tested. Chromium, zinc, and bromine also show differences between the two groups at a probability exceeding 0.95. Variations are indicated in scandium and cesium also, but are less strongly supported. Hafnium and selenium show no significant differences.


Competitive enzyme-liked immunoassay with use of soluble enzyme/antibody immune complexes for labeling. I. Measurement of human choriogonadotropin. We described the principle of a new enzyme-immunoassay, competitive enzyme-liked immunoassay (CELIA), for quantitative measurement of soluble antigens and haptens. In the assay, binding of antibody to antigen-immunosorbent is competitively inhibited by the free antigen to be measured. The amount of first antibody bound to the immunosorbent is measured by an enzymatic technique in which a heterologous bridging antibody and a soluble antibody/enzyme immune complex are applied in sequence. The soluble complex we used was rabbit antiperoxidase/horseradish peroxidase. Peroxidase activity is inversely proportional to the concentration in the original sample of the substance to be assayed. The enzyme-linked reagents are potentially widely applicable to any substance to be measured. To demonstrate the feasibility of CELIA, we report a preliminary study of its application to the measurement of human chloriogonadotropin in serum and urine. The assay described for this hormone has a working range of 1 to 50 int. units per milliliter of sample. The technique obviates the disadvantages associated with measurement and handling of radioisotopes in radioimmunoassays and the only major instrumentation required is a centrifuge and a conventional spectrophotometer.


Measurement of glucose in plasma, with use of immobilized glucose oxidase and peroxidase. We describe a novel fluorometric technique for simple, rapid, and economical assay of glucose by use of immobilized glucose oxidase and peroxidase. A cylindrical magnetic stirrer was specially designed to hold the immobilized enzymes firmly and to allow the reaction minzymic transformation quickly. Blood plasma can be assayed directly with no pretreatment. Ascrobic acid, uric acid, and creatinine in concenctrations of 0.25, 0.25, and 0.2 g/liter, respectively, did not interfere. The linearity of the assay was extended to 4.0 g of glucose concentrations. Results by our technique and by the otoluidine or hexokinase methods agreed well. The immobilized enzymes are stable for several months and can be used for several hundred highly accurate and reproducible assays.


Factors influencing results for thyroxine concentration in blood, as measured in paper filter spots in a screening program for neonatal hypothyroidism. To reduce the number of false-positives in our screening program for neonatal hypothyroidism, we investigated the effect of age, body weight, and interval between sampling and analysis on results for filter paper spot thyroxine determination. The statistical analysis included an analysis of covariance. The age of the infant did not influence the results, the interval influenced them minimally, but weight did require a correction factor of about 0.16 ng/kg below the mean weight. This correction factor should reduce our false-positives from 0.75% to 0.60%. Furthermore, because all hypothyroid infants had results that fell more than 2.8 SD below the geometric mean, we intend to measure thyrotropin concentrations in the blood eluate of all the spots with a thyroxine concentration below that cut-off point.


Improved continuous-flow method for detemination of total serum hexosamines. We describe an automated determination of serum hexosamine by the Elson-Morgan reaction, together with reagent modifications that minimiz interference from amino acids and sugars present in acid hydrolysates of sera. We used a novel 15-min autoclave procedure for the acid hydrolysis of sera before analysis to facilitate the detemination as compared to the 4-h hydrolysis used in the conventional manual method. Results correlated well (r = 0.906) with those obtained by the corresponding manual method.


Simplified radioimmunoassay for triiodothyronine: evaluation of a new commercial kit. A new triiodothyronine (T3) radioimmunoassay kit offers simplicity of method, short incubation time, specificity, and reproducibility. 125I-labeled T3 and unknown serum (or standard) are added to alkaline buffered (pH 12.4) Sephadex G-25 columns; the T3 is freed of proteins with an alkaline buffer wash, antiserum is added, and the columns are incubated at room temperature for 2 h. "Free" hormone (i.e., unbound to antiserum) remains on the column after a second wash. The radioactivity on the column is counted and related to the total activity added. The T3 standards produced a straight line on log-logit plot; reactivity of analytical-grade thyroxine was 0.6% on weight basis; and the analytical recovery of T3 added to a T3-free serum was complete. Euthroid, hyperthyroid, and hypothyroid ranges were 0.74-2.64, 2.91-7.52, and 0-1.21 mug/liter, respectively. These values correlated well with both the clinical status of patients and with values obtained by an established radioimmunoassay method. The procedure is simply done; results may be obtained in less than 4 h.


Creatine kinase isoenzymes in cerebrospinal fluid in a case of brain damage. We present a case of a 11/2-year-old boy with toxic enteritis, consecutive consumption coagulopathy, and sever brain damage. During the acute phase we found high activity of the BB isoenzyme of creatine kinase in cerebrospinal fluid, but not in the serum. Isoenzyme MM could also be found in the spinal fluid (37.9% of the total activity). We conclude that analysis for creatine kinase isoenzymes in spinal fluid is of clinical importance.


A syndrome of generalized elastic fiber deficiency with leprechaunoid features: a distinct genetic disease with an autosomal recessive mode of inheritance. Three male infants with generalized elastolysis and leprechaunoid features from two related and consanguineous parents of Italian origin died in the first year of life following severe cardio-pulmonary complications. While these children showed a decrease in elastic fibers, no degeneration was noted and histochemical as well as systemic metabolic studies were negative. It is postulated that this disease is a variant of cutis laxa or at least that the absence of granular degeneration of the elastic fibers described by Goltz is a secondary manifestation present only in older children. Prenatal diagnosis of this syndrome is not yet possible since no intracellular or biochemical changes have been identified. In view of the familial occurrence of this syndrome, and the association of specific clinical and pathological findings, we suggest that we are dealing with a distinct hereditary disorder of the connective tissue.


A rapid micromethod for prenatal diagnosis of Lesch Nyhan syndrome. A method is described which enables prenatal diagnosis of Lesch Nyhan Syndrome (HGPRT deficiency) to be made within 7-10 days. The procedure is based on the direct cultivation of amniotic cells in microtest II plates; the HGPRT reaction is performed in individual wells containing between 500 to 10,000 cells, and is followed by separation of the radioactive reaction products by means of microchromatography on 3 cm x 5 cm PEI plates. This method permits determination of the actual HGPRT enzyme activity of the cell lines.


Frequency of the carrier state for X-linked chronic granulomatous disease among females with lupus erythematosus. Carriers for chronic granulomatous disease (CGD) and patients with lupus erythematosus (LE) share several characteristics: They are mostly females, they reduce nitroblue tetrazolium (NBT) poorly in their neutrophils, and, in some cases, they have similar skin lesions. We thus investigated 19 female LE patients for the presence of laboratory findings characteristic of the carrier state for CGD. None of these patients turned out to have the combined abnormality of neutrophil bactericidal activity and neutrophil NBT-reduction that is diagnostic of CGD carrier state in the X-linked form. An increased frequency of CGD carriers among female LE patients thus appear to be unlikely. Why some CGD carriers develop skin lesions typical of LE remains unexplained.


Partial trisomy 15 in a young girl.

Partial trisomy 15 in a young girl. Severe mental retardation, growth failure, microcephaly, micrognathia, cleft palate, hooked nose, low-set thin flabby ears, pronounced elongated philtrum, and generalized decreased subcutaneous tissue characterized a 5 year 9 month old female with karyotype 47,XX,+15,del(15)(q15).


The myogenic scapulo-peroneal syndrome. Muscular dystrophy in the K. kindred: clinical study and genetics. Additional study was carried out of six generations belonging to the K. kindred, which were previously investigated by Oransky (1927). Eighteen members of this kindred were studied. In the early stages of the disease a sharp dissimilarity was observed in the phenotypes in the K. kindred, resulting from different rates of development and varying intensity in the course of the disease, so that it ranges from evident scapulo-peroneal amyotrophy to cases resembling Landouzy-Déjerine muscular dystrophy. In the later stages of the disease the clinical data on those affected were noted to be very similar. Muscular dystrophy in the K. kindred has been inherited as an autosomal-dominant type for five generations. The penetrance of the gene is almost complete. The expressivity of the gene varies from abortive to severe forms of disease. From the clinical and genetic data available at the present time, is seems that the muscular dystrophy in the K. kindred is one of the varieties (namely, a descending type with a "jump") of the facio-scapulo-limb (or facioscapulohumeral) muscular dystrophy. The scapulo-peroneal syndrome could be a long stage in the development of the disorder in some members of the K. kindred. As well as muscular dystrophy, some muscle anomalies have been independently inherited in the K. kindred. Apparently, muscular dystrophy and muscle defects are caused by the non-allele genes; this ensures the independent distribution of these signs among the members of the K. kindred.


Familial pericentric inversion of chromosome 19, inv(19) (p13q13) with a note on genetic counseling of pericentric inversion carriers. An inmate of a hospital for the mentally retarded was found during a cytogenetic screening programme to have karyotype 46,XY,inv(19)(p13q13)mat. Clinical, cytogenetic and family findings are presented and it is concluded that the chromosomes abnormality was probably not the cause of the patient's retardation. The problem of genetic counseling of inversion carriers is examined in some detail and estimates of risk are given.


Lymphocyte transformation test with liver-specific protein and phytohaemagglutinin in patients with liver disease. Lymphocytes from thirty-four untreated patients with various liver diseases were stimulated in a lymphocyte transformation test with liver-specific protein (LSP). Eight of ten patients with chronic active or persistent hepatitis, two of five patients with non-alcoholic cirrhosis and six of nineteen patients with acute hepatitis showed a positive in vitro reactivity to LSP. In a control group of twelve persons without evidence of liver disease, eleven gave a negative response to LSP stimulation, whereas one person showed a positive response. Among fourteen patients with chronic hepatitis or non-alcoholic cirrhosis treated with prednisone at the time of the investigation, only one showed reactivity to LSP stimulation. Three patients in this group had previously had a positive reaction before prednisone was given. There was no statistically significant correlation between the reactivity to LSP stimulation and the presence or absence of hepatitis-associated antigen (HBAg) in serum, or with the biochemical liver parameters. The response to in vitro stimulation with phytohaemagglutinin (PHA) was found to be significantly lower as compared with the control group in eleven patients with alcoholic liver disease and in the patients with acute hepatitis who had HBAg in serum. This decrease in reactivity could apparently not be ascribed to immuno-suppressive factors in the patients' sera.


Immune complex infusion in the rat. Renal functional and morphological changes. Immune complexes were precipitated from pooled goat antisera to human Cohn Fraction II. After washing the complexes were solubilized in antigen excess. When complexes containing 10 mg of antibody were infused into saline-loaded rats an episode of oligoanuria lasting 4-6 hr resulted followed by transient proteinuria. By 24 hr after complex infusion protein excretion had returned to pre-infusion levels. With infusion of 5 mg of antibody similar functional alterations, though of lesser severity, occured. Control animals infused with Cohn fraction II alone in an amount and protein concentration equal to the experimental groups exhibited none of these features. Changes in blood pressure did not occur. Immune complex deposition was not detectable in the glomeruli by immunofluorescence microscopy. A mild microangiopathy affecting mainly interstitial vessels, with some glomerular endothelial cell swelling, was evident on light microscopy. It is suggested that circulating factors resulting from complex infusion rather than glomerular complex localization account for the changes observed.


The effect of respiratory immunization on cell-mediated immune effector cells of the lung. In order to understand the mechanisms of cellular immune injury in hypersensitivity pneumonitis, the effect of type of antigen on cell-mediated immunity in guinea-pigs receiving respiratory immunization was studied. Lymphocytes obtained by pulmonary lavage were compared with those from peritoneal exudate following immunization with either a soluble protein, human serum albumin, or a particulate suspension of Thermoactinomyces vulgaris. Assays were obtained without mixing cells from these two sources. Statistically significant increases (13-22%) in the number of alveolar rosette-forming cells (RFC) were found in the animals immunized with either antigen, but only the particulate T. vulgaris was also capable of inducing a systemic increase of such cells. That this increase in RFC could be due to specifically reactive lymphocytes was demonstrated by the production of antigen-stimulated macrophage migration inhibition. Some evidence was obtained that indicated that T. vulgaris could act both as a non-specific B-cell stimulant and a specific T-cell activator. The concept of a hypothetical pulmonary 'barrier' is discussed which must be overcome to induce systemic immune responses following respiratory immunization. T. vulgaris must be added to the list of known agents or means for overcoming this 'barrier'.


Modification of cutaneous leishmaniasis in the guinea-pig by cyclophosphamide. Pretreatment of guinea-pigs with cyclophosphamide (Cy) (300 mg/kg) 3 days before cutaneous infection with Leishmania enriettii caused an increased intensity of the lesion at the site of infection and an increase in the incidence of widespread metastases. Decreased levels of circulating antibody were found from the first to fourth week after infection. Decreased delayed type hypersensitivity could only be detected beyond 4 weeks. Peritoneal macrophages obtained form guinea-pigs 3 days after Cy pretreatment showed increase rather than decreased ability to phagocytose L. enriettii. Phagocytosis of L. enriettii by peritoneal macrophages obtained from guinea-pigs pretreated with Cy 24 days previously was normal. It is suggested that more attention should be taken of antibody levels during the early phase of infection and that control of infection could be due to a synergism between antibody and cell-mediated immunity. Upset in the balance by suppression of either function might lead to the development of widespread metastatic lesions.


Complement activation in patients with Gambian sleeping sickness. Low levels of complement components C3, C4 and factor B were found in the sera of most patients with early and advanced Gambian trypanosomiasis. C3 levels increased after treatment with melarsoprol (Mel B). Nearly all patients had high serum IgM levels and a significants correlation was found betwen high serum IgM levels and low serum C3 levels. High concentrations of cryglobulins, which contained IgM and C3, were found in most of sera tested. Formation of large molecular weight IgM complexes may play an important part in producing the low C3 levels found in patients with Gambian sleeping sickness.


Antigenic diversity among Onchocerca volvulus in Nigeria, and immunological differences between onchocerciasis in the savanna and forest of Cameroon. Onchocerciasis is a remarkably chronic infection and, in West Africa, there are distinct clinical and pathological differences between the disease in the savanna and in the forest. Experiments were carried out to see whether antigenic diversity among Ochocerca volvulus worms might contribute to these features. Extracts of adult worms and or microfilariae were analysed by disc electrophoresis on polyacrylamide gels and showed a remarkable diversity of protein patterns but no consistent savanna/forest or Nigeria/Cameroon differences. Indirect haemagglutination using cross-absorbed sera was used to look for antigenic diversity or identity between different O. volvulus worms. Sera of patients were absorbed with antigens extracted from worms taken from those paitents and then titrated against sheep erythrocytes coated with the same antigens. It was shown that an adult Onchocerca volvulus could be either antigenically identical with, or distinct from, another worm taken from the same patient. In one Nigerian village identity was shown in two instances nad diversity in five. Microfilariae lack some antigens possessed by adult worms. This test has shown consistent savanna/forest differences in worm antigen patterns and antibody responses. In order to explain the pattern of these differences it was necessary to postulate that forest patients possessed antibodies to another worm which had antigens in common with savanna Onchocerca. The relevance of these findings to the pathology and prevention of onchocerciasis is discussed.


Soluble factors in immune sera of mice. I. Specific and non-specific suppressive activity. Serum collected from mice 7 days following immunization with heterologous erythrocytes (SRBC, HoRBC) contained potent and specific immunosuppressive activity. When the serum was filtered through a Diaflo UM-10 membrane the activity was recovered in the filtrate, indicating that a factor less than 10,000 Daltons is responsible for it. Filtrates obtained from animals immunized with soluble antigens (BSA, POL) in FCA suppressed the 7S response to SRBC but had no effect on the 19S response. This non-specific suppressive effect on the7S response was also shown by filtrates obtained from sera of animals injected with FCA or other adjuvants (LPS, poly A:U). The 19S response to SRBC was either not affectd (LPS, poly A:U) or actually enhanced. (FCA). The SRBC-induced filtrates markedly suppressed the espression of DH to SRBC.


Treatment of Goodpasture syndrome with cyclophosphamide, prednisone and plasma exchange transfusions. Repeated plasm exchanges were performed in a 44-year-old man with Goodpasture syndrome, also treated with cyclophosphamide and prednisone. Improvement was observed within 3 weeks of starting the protocol, and by the 76th week, endogenous creatinine clearance had increased from 30 to 56 ml/min/1.73 M2 and serum albumin from 2.7 to 3.7 g/dl. Prior treatment with immunosuppressive drugs had not significantly influenced circulating antibody levels. But sustained suppression of antibody was achieved after the plasma exchanges were begun, suggesting that physical removal of circulating antibody combined with antiproliferative drug treatment may be a useful way to control undesirable humoral immune responses.


Comparison of allergic aspermatogenesis with that induced by vasectomy. I. In vivo studies in the guinea-pig. Groups of male and female guinea-pigs were immunized with homologous epdidymal sperm in Freund's complete adjuvant (FCA) and skin tested at weekly intervals with a heat-treated extract of the sperm or with PPD. In females, the skin response to both antigens was similar to that to any standard protein antigen. In males, the response to sperm extract  varied with the induced auto-immune orchitis, reaching a maximum immediately after testis lesion was most severe and as recovery was beginning (shown histologically): the response to PPD was decreased at 1 week after immunization, but subsequently was similar to that in females. Skin tests on guinea-pigs 8 months after bilateral vasectomy (when both ends of vasa were ligated), showed no evidence of delayed hypersensitivity to sperm, although there was marked histological evidence of reduced spermatognesis, due to back pressure atrophy.


Accessibility to antibodies of acetylcholine receptors in the neuromuscular junction. Antibodies to the acetylcholine receptor are present in the serum of myasthenic patients but one does not know if, in vivo and in situ, they can penetrate the intact neuromuscular junction and block directly the receptor. The present experiments demonstrate that molecules the size of antibodies can reach acetylcholine receptor in situ. The mouse diaphragm with its intact neuromuscular junction was used as a source of acetylcholine receptor. The receptor was revealed either directly by iodinated alpha-bungarotoxin covalently coupled to IgG or indirectly, once labelled with cobra toxin, by iodinated anti-cobra toxin antibodies.


Deaths due to childhood asthma. Are they preventable? Seven cases of death in patients with severe asthma are presented. The cause of death in asthmatic children is discussed under the following headings: mucus plugs, insufficient treatment, infection, aspiration, concomitant disease, oversedation, theophylline toxicity, overuse of metered dose pressurized aerosols, inadequate steroid therapy, and sudden cardiopulmonary arrest. The authors conclude that asthmatic deaths are preventable.


Treatment of neoplasia of the bladder with topical application of a synthetic peptide complex preparation: preliminary results. Preliminary results are reported on the endo-vesical use of a complex of synthetic peptides ('Peptichemio') in 15 patients suffering from bladder neoplasia who were judged to be beyond surgery or radiotherapy. The agent was introduced into bladder in 20 mg doses diluted in 50 ml of 5% glucose and retained for approximately 30 minutes. Treatment was repeated every 5 to 7 days. The disappearance of the neoplastic mass in 13% of the cases and its reduction in a further 53% encourage the continued use of the preparation in local regional therapy of bladder neoplasia, and the findings help to establish treatment times and doses. For the moment, the authors recommend increasing the interval between treatments to 10 days or more once the neoplastic mass has been reduced or disappeared.


The effects of the sub-chronic administration of an anti-histamine, clemastine, on tests of car driving ability and psychomotor performance. A double-blind placebo-controlled crossover trial was carried out in 21 normal volunteers to study the effects of sub-chronic administration of clemastine on car driving ability and psychomotor performance. Subjects were assessed on 5 tests representing various aspects of everyday car driving experience, and on subjective rating scales for mood, sleep, and psychomotor integration. The results showed that repeated doses of 1 mg clemastine b.d. for 3 days had no significant consistent effect on any of the parameters measured.


Betamethasone-I7, 2I-dipropionate ointment: an effective topical preparation in lichen ruber planus. A clinical evaluation was carried out in 19 patients with lichen ruber planus to assess the effectiveness of 0.05% betamethasone-17,21-dipropionate ointment in the treatment of patients whose lesions had developed resistance to prolonged treatment with 0.1% betamethasone-17-valerate ointment. The new steroid ointment was substituted for the previous preparation and applied at the same frequency, once or twice daily, for 2 to 3 weeks. The results at the end of the betamethasone-17,21-dipropionate period were classified as better in 14 (73.7%) and equal in the remaining 5 patients.


Local treatment of psoriasis of the scalp with clobetasol propionate in alcoholic solution: a comparison of once and twice a day application. A clinical evaluation was carried out in 53 patients with moderate to severe psoriasis of the scalp to assess the effectiveness of a 0.05% alcoholic solution of clobetasol propionate. Patients were treated on a non-selective basis with either a once a day or twice a day application. After a 2-weeks' treatment period, the results showed that there was an excellent or good response in 65% of patients treated once a day and in all of the patients on twice daily application of the topical steroid. It is recommended, therefore, that clobetasol propionate should be applied twice daily for the control of severe cases of psoriasis of the scalp.


Treatment of chronic gastric ulcer with gefarnate: a long-term controlled therapeutic trial. A long-term controlled therapeutic trial of gefarnate (600 mg daily) in the treatment of chronic gastric ulcer was carried out in 32 patients. Assessment was based on the clinical course and on repeated radiological and endoscopic examinations, which were performed after 3-months' treatment, at the end of the 1-year trial period, and towards the end of a follow-up period of approximately 2 1/2 to 3 1/2 years. In men with gastric ulcer, gefarnate proved to be highly effective, with 10 out of the 11 patients who received this treatment showing persistent healing of the ulcer compared with only 4 out of the 11 who were on dummy treatment. In the very small group of women it was not possible to show any difference between treatments. No adverse reactions to gefarnate occurred. These results suggest that gefarnate is a safe and effective treatment for chronic gastric ulcer.


Clinical evaluation of clobetasone butyrate in the treatment of children with atopic eczema, and its effect on plasma corticosteroid levels. Studies were carried out to assess the effectiveness of clobetasone butyrate in treating eczema and psoriasis, and to determine if the compound had any effect on plasma cortisol levels. In the first trial, 71 children with bilateral symmetrical atopic eczema lesions were treated twice daily for 1 week, on one side with 0.05% clobetasone butyrate cream or ointment and on the other with 0.0125% flurandrenolone cream or ointment. Lesions improved or healed in the majority of the patients. Treatment preference showed a trend in favour of clobetasone butyrate but the difference was not statistically significant. In a second open trial, 29 adults with eczema or psoriasis were treated twice daily with clobetasone butyrate for 1 week: lesions in 10 patients remained static, 12 improved, and 7 were cleared. Plasma corticosteroid levels remained within the normal range at the end of the treatment period.


Pachytene mapping of the male Chinese hamster. Minced seminiferous tubules of male Chinese hamsters when treated with a mixture of trypsin (one part) and  McCoy's 5a growth medium with 20% fetal calf serum (nine parts) at 4 degrees C, washed twice with the regular growth medium, incubated at 37 degrees C in growth medium for 4 h, and harvested without Colcemid and hypotonic pretreatments, gave excellent pachytene morphology for studies on chromomere patterns. The Giemsa banding patterns of all somatic metaphase chromosomes except the sex chromosomes of the hamster cells correspond well to the chromomere patterns of meiotic pachytene bivalents.


The in vitro isolation and characterization of monosomic sublines derived from a Colcemid-treated Chinese hamster cell population. A series of 18 sublines deficient for one or two chromosomes was isolated in vitro from a pseudodiploid Chinese hamster cell population treated appropriately with Colcemid to obtain maximal induction of aneuploidy. Chromosomes 10 and 11 were the most frequently involved and, to a lesser extent, chromosome 9 and the partially deleted 6. Since analysis of chromosome banding patterns did not reveal any evidence of chromosome rearrangement, it was concluded that entire chromosomes were absent, and thus the sublines are presumed to be monosomic. The monosomic state was quickly lost during serial culture, and the 15th passage after isolation, the monosomic cell type accounted for less than 50% of the cell population in over half the sublines. This was due to a shift in chromosome number back to 22, apparently by reacquistion of the absent chromosome, or to the appearance of tetraploidy.


Further evidence of X-linkage of hypoxanthine phosphoribosyl-transferase in the mouse. A variant of the HPRT-A9 mouse cell line was fused with wild type diploid mouse bone marrow cells to obtain an HPRT+ line. The unique chromosomal features of the A9 parent, including the presence of a t(X;3) translocation and the absence of normal chromosomes 15,16, 17, 18, and the X, have permitted use of this intraspecific hybrid for chromosome mapping. Back-selection of hybrid cells in 8-azaguanine for loss of HPRT resulted in the loss of the Xchromosome derived from the diploid parent, providing evidence of the X-linkage of the HPRT locus.


A mitotically unstable human dicentric Y chromosome in a male pseudohermaphrodite. A patient with a mitotically unstable dic(Y)(p11) chromosome is reported. Physical examination revealed a small penis with severe hypospadia, undescended testes, rudimetary vagina, uterus, left fallopian tube, and no stigmata of Turner syndrome. Longitudinal chromosome studies over a four-year period, including blood, skin, foreskin, and testicular tissue, revealed 45,X/46,X,dic(Y)(p11)/46,X,del(dic Y) mosaicism. The proportions of these cells varied in the different tissues, and only 45,X and 46,X,del(dic Y) were major cell lines in testicular tissue. Additional minor cell lines were present mainly in peripheral blood: 47,X,dicY,dicY; 47,X,dicY,del(dicY); and 47,X,del(dicY),del(DICY). Premature disjunction of one of the centromeres in a high percentage of the dicentric Y chromosomes in metaphase was observed by Q- and C-banding. Lymphocytes at anaphase and telophase showed lagging Y chromosomes, fragments, and nondisjunction. These observations indicate a high degree of mitotic instability and thus raise the question of the effect of premature centromeric disjunction on mitotic instability of dicentric chromosomes.


Does the T-locus in the mouse include ribosomal DNA? The proportion of rDNA in the mouse genome is shown to have no systematic relation to the T-locus genotype, and hybridization in situ with 125I-RNA does not label chromosome 17, which carries the T-locus. Thus rDNA need no longer be taken into account in explaining the unique properties of the alleles at this locus. The number of genes for rRNA precursor averaged 153 per haploid genome, but a large measure of individual variation is suggested.


The relationship of a specific chromosomal region to the development of the acrosome. In early spermatids of Urodeles the chromosome segments bearing constitutive heterochromatin are localized in one half of the round nucleus; this region becomes the basal part of the long nucleus of the spermatozoon. The euchromatic chromosome segments extend toward the anterior nuclear pole in a bouquet configuration (Macgregor and Walker, 1973). In the course of spermiohistogenesis, one of the heterochromatic regions (the acrosomal chromocenter) migrates from the basal part to the anterior half of the spermatid nucleus. This heterochromatic block is identical with a species-specific, definite C-band in the karyotype. This relationship between the acrosomal chromocenter and a specific chromosomal C-band was established in Triturus cristatus, T. marmoratus, T. alpestric and Cynops pyrrhogaster. In closely related species this particular C-band lies on similar chromosomes. - While the spermatid nucleus still retains its round shape the acrosomal chromocenter despiralizes into a long heterochromatic thread (acrosomal thread). Precisely at the position of this thread the nucleus evaginates and acquires a pear-like shape. During the elongation of the nuclear protrusion the acrosomal thread remains associated with the anterior end. At termination of spermiogenesis it lies closely below the acrosome in the tip of the spermatozoon. Spontaneous aberrations which affect the acrosomal chromocenter or the thread lead to the development of spermatozoa with defective tips. - Several euchromatic segments, interspersed between the heterchromatic segments, can be recognized in the completely despiralized acrosomal thread. Genes responsible for the morphogenetic activities of both, the acrosomal chromocenter and the acrosomal thread, in the development of the spermtip, might be localized in these interspersed euchromatic segments. The existence in higher vertebrates of an acrosomal chromocenter or an equivalent chromosomal region is discussed.


Meiotic drive for B-chromosomes in the primary oocytes of Myrmeleotettix maculatus (Orthopera: Acrididae). Using a modified technique which allowed observation of chromosome orientation in the primary oocyte of grasshoppers at the onset of anaphase, it has been possible to establish that the B-chromosomes is distributed preferentially on the egg side of the metaphase plate rather than the polar body side. The frequency of this preferential orientation matches very closely the level of preferential transmission determined from breeding experiments using individuals from the same population. The spindle is asymmetrical in the primary oocyte of this species, and a possible explanation of the meiotic drive is proposed as a result of the conical shape of nucleoplasm surrounding this spindle. The autosomal chiasma frequency of these females is generally lower than comparable males and is increased by the presence of B chromosomes; but the ciasma frequencies of the sexes respond differently to the addition of 1 and 2 B-chromosomes.


Cytogenetics of two species of Euceraphis (Homoptera, Aphididae). Somatic cell divisions, spermatogenesis, and the prophase stages of primary oocytes, are described for two species of birch aphid, Euceraphis betulae (Koch) and E. punctipennis (Zettersted). Females of E. betulae have two autosome pairs, two pairs of X-chromosomes of different lengths, and two B-chromosomes. Females of E. punctipennis have the same number of X-chromosomes. The sex determination system is X1X2O. E. punctipennis males sometimes have only one B-chromosome. In the spermatogenesis of E. Betulae, pairing of homologous autosomes occurs in early prophase I, but no evidence was found of chiasmata or end-to-end alignment of homologues. Instead, homologues remain closely aligned in parallel as they condense into metaphase, and anaphase I separates the products of pairing in a strictly reductional manner. The two unpaired X-chromosomes and both B-chromosomes are stretched on the anaphase I spindle and all four pass into the larger secondary spermatocyte. The second division is equational. The B-chromosomes thus show accumulation in spermatogenesis, which must be compensated in some way by an elimination mechanism in oogenesis. Meiosis of E. punctipennis is highly anomalous. The two autosomes pair but separate again in early prophase I, then one homologue becomes heterchromatic and is apparently rejected from the late prophase nucleus. A single, equational maturation division follows. In female neiosis I, both species show highly characteristic diplotene figures with multiple chiasmata, the B-chromosomes remaining unpaired. These results are discussed in relation to previous work on aphid cytogenetics.


Peroxidase from human cervical mucus. The isolation and characterisation. A peroxidase has been isolated from pooled specimens of human cervical mucus. Investigation of substrate and inhibitor specificity, pH optimum, chromatography on Sephadex G-200, disc electrophoresis, ultraviolet absorption spectrum and the effects of temperature and ionic strength variation are described. A study of the steady-state kinetics is reported using hydrogen peroxide and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) as substrates and a catalytic mechanism is proposed.


Characterisation of the lysosomal heterogeneity in Chinese-hamster fibroblasts. 1. Homogenates of Chinese hamster fibroblasts have been fractionated by analytical centrifugation techniques. Lysosomes have been characterised and identified by the distinctive behaviour of specific marker constituents. 2. The acid hydrolases of hamster fibroblasts were located in two distinct organelles, characterised by differences in their equilibrium densities and rates of sedimentation. The lighter population of lysosomes with a modal density of 1.10 g cm-3, contained glycosidases, nucleases and acid phosphatase. The denser population, with a modal density of 1.15 g cm-3, contained proteases and arylsulphatase B. 3. Acid hydrolases in both populations showed similar degrees of latency and structure-linked sedimentability. 4. The heterophagic activity of hamster fibroblasts was investigated by growing the cells in the presence of [14C]sucrose, [14C]inulin and Triton WR 1339. These compounds are taken up by the cells and appear to be located in the denser population of lysosomes. 5. The low density of the lighter population of lysosomes appears to result from a high content of cholesterol esters and triacylglycerols. It is suggested that the lysosomes may be the site of storage of a considerable proportion of these lipids in cultured hamster cells.


The initiation of globin synthesis in differentiating rabbit-bone-marrow erythroid cells. At a particular stage of erythropoiesis, the immature erythroid cell commences synthesis of the messenger RNA for globin. Experiments were performed to determine whether this mRNA is immediately translated into globin or whether there is any substantial delay and accumulation of mRNA awaiting a further stage of differentiation. Rabbit bone marrow erythroid cells were separated according to their degree of maturity by velocity sedimentation on a sucrose gradient. Their relative abilities to synthesise globin were measured by radioactive labelling in culture and the relative proportions of globin mRNA in cellular RNA were determined by hybridisation to DNA complementary to globin mRNA. The increasing ability to synthesise globin closely parallelled the concentration of globin mRNA at all stages of differentiation, thus suggesting that there is no significant delay between transcription and translation.


The mechanism of the synthesis of indoleglycerol phosphate catalyzed by tryptophan synthase from Escherichia coli. Steady-state kinetic studies. The mechanism of indoleglycerol phosphate synthesis from indole and D-glyceraldehyde 3-phosphate catalyzed by tryptophan synthase has been investigated by steady-state kinetic techniques. The equilibrium constant and the progress curves were measured by use of the difference in absorbance between indole and indoleglycerol phosphate. Stopped-flow measurements show that only the non-hydrated form of D-glyceraldehyde 3-phosphate serves as substrate. The product analogue indolepropanol phosphate was used as an inhibitor to discriminate between possible mechanisms. The data agree well with an ordered addition mechanism with D-glyceraldehyde 3-phosphate adding first. Mechanisms involving random addition of substrates or ordered addition with indole adding first can be excluded because indolepropanol phosphate is a competitive inhibitor only towards glyceraldehyde 3-phosphate. The high affinity of tryptophan synthase for indoleglycerol phosphate leads to product inhibition even at small extents of reaction. Glyceraldehyde 3-phosphate combines with the enzyme with an apparent second-order rate constant, which is not diffusion controlled and generates a site with high affinity for indole.


Steady-state kinetic studies of the synthesis of indoleglycerol phosphate catalyzed by the alpha subunit of tryptophan synthase from Escherichia coli. Comparison with the alpha2 beta2-complex. For the alpha subunit of tryptophan synthase and at constant concentration of D-glyceraldehyde 3-phosphate the saturation curves with respect to indole concentration are weakly sigmoidal. This phenomenon can be explained by interaction between indole bound to the effector site established previously and the active center of the monomeric alpha subunit. Kinetic studies of the inhibition of indoleglycerol phosphate synthesis by the analogue indolepropanol phosphate show that the inhibition is competitive with respect to D-glyceraldehyde 3-phosphate and non-competitive with respect to indole. Mechanisms with random addition of substrates or ordered addition with indole binding first can therefore be excluded. A quantitative fit of the data has been obtained to an ordered addition mechanism with D-glyceraldehyde 3-phosphate binding first and with a distribution of the enzyme between two states differing in V, governed by the binding of indole to the effector site. The kinetic constants obtained for the alpha subunit have been compared with those of the alpha 2 beta 2 complex of tryptophan synthase. Protein-protein interaction of the alpha subunit with the beta 2 subunit (a) does not alter the catalytic of the indoleglycerol phosphate synthesis, (b) suppresses the substrate activation by indole, and (c) changes the various equilibrium, rate and steady-state constants in the sense of conveying higher substrate specificity and catalytic efficiency to the alpha-subunit. The occurrence of local and gross conformational changes in the tryptophan synthase system is discussed.


Interaction of RNA polymerase from Escherichia coli with DNA. Effect of temperature and ionic strength on selection of T7 DNA early promoters. Factors influencing promoter site selection by Escherichia coli RNA polymerase were investigated using T7 DNA as template. The utilization of the three major early promoters A1, A2 and A3, was followed by processing the RNA transcripts with RNAase III, which generates the three corresponding initiator RNA fragments. The three promoters proved to be functionally distinct. A strong differential effect of temperature and ionic strength on promoter selection was observed. The transition temperature of promoters A1, A2 and A3 was measured directly by preincubating, at different temperatures, RNA polymerase and T7 DNA, with primer-substrate combinations which selected each site independently. The transition temperature of the three sites was markedly different. Promoter A3 was used predominantly at low temperature, whereas A1 or A2 promoters were gradually activated by increasing the temperature. The temperature response curves were strongly dependent upon the salt concentration. On the other hand, challenge experiments with rifampicin or poly (inosinic acid) showed that, once preinitiation complexes are formed by incubating RNA polymerase with DNA at 37 degrees C, the three sites A1, A2 and A3 promote chain initiation with equal efficiency and at the same rate.