Effects of Gaseous CO2 and bicarbonate on the growth of Neisseria gonorrhoeae. The quantitative and qualitative growth response of Neisseria gonorrhoeae strains was assessed under the following conditions: incubation in a candle jar (approximately 2.5% ambient CO2) on medium without bicarbonate, incubation in air on medium without bicarbonate, incubation in air on medium with bicarbonate, and incubation in air in a sealed jar on medium with bicarbonate. Incubation in the presence of ambient CO2 (candle jar) resulted in the highest plating efficiencies for the three laboratory strains 9, 62, and 2686. The addition of NaHCO3 to the medium enhanced the growth response in air of all three strains, particularly if incubation was carried out in a closed environment (sealed jar). The qualitative growth response of 34 clinical isolates and laboratory strains was assessed under the same conditions of incubation after the plating of an inoculum containing approximately 2 X 10(6) bacteria. The strains were divided into different classes based on their growth responses. About 40% of the strains grew as well on bicarbonate-containing medium incubated in air in sealed jars as they did on medium without bicarbonate incubated in a candle jar. Ten percent of the strains showed only slight growth on bicarbonate containing medium incubated in sealed jars and appeared to have an almost obligate requirement for ambient CO2. Twenty percent of the strains apparently had partially lost their requirement for gaseous C02 and showed slight growth in air on medium without bicarbonate and slight to moderate growth in air on medium containing NaHCO3. The remaining 30% seemed to have completely lost their requirement for gaseous C02 and/or the bicarbonate anion and grew almost as well in air on medium without bicarbonate as they did in either ambient C02 (candle jar) or on medium containing bicarbonate incubated in a sealed jar. These results suggest that N. gonorrhoeae strains may vary widely in their requirements for CO2 and/or the HCO3-anion. Incubation in the presence of ambient CO2 tends to maximize the growth response on solid medium of those strains, which require it for growth. The presence of ambient CO2 is particularly important if growth is to be obtained after the plating of small inocula. Medium conatining 0.1% NaHCO3, if incubated in a closed environment (sealed jar), apperas to be equivalent to medium without bicarbonate incubated in ambient CO2 in supporting the growth of some but not all strains of N. gonorrhoeae.


Screening for cross-reacting capsular polysaccharide K antigens of Escherichia coli using antiserum agar. Agar plates containing antiserum against group B meningococcus or Haemophilus influenzae type b were used to determine the prevalence of cross-reacting K1 and K100 capsular polysaccharide antigens in 265 isolates of disease-causing Escherichia coli. K1 antigen was found in 22% of isolates from various sites. K100 antigen was found in only three isolates. This technique is a convenient method to detect specific E. coli K antigens for evaluation as possible factors important in the virulence of the organism.


The metabolism of IgE in patients with immunodeficiency states and neoplastic conditions. Serum IgE concentrations were determined and IgE turnover studies were performed in control individuals as well as in patients with several disease states. Patients with common variable hypogammaglobulinemia, thymoma and hypogammaglobulinemia, ataxia telangiectasia, and selective IgA deficiency had significantly decreased mean serum IgE concentrations. In turnover studies, this was found to be due to decreased IgE synthesis. In spite of these depressed mean values, some patients with common variable hypogammaglobulinemia had normal serum IgE concentrations and synthetic rates. Patients with the Wiskott-Aldrich syndrome had a significantly elevated mean serum IgE concentration. In one of four patients studied with the turnover technique, a strikingly high IgE concentration was present and was associated with an elevated IgE synthetic rate. Three other patients had both normal serum IgE concentrations and synthetic rates. Patients with chronic lymphocytic leukemia had significantly decreased mean serum concentrations and synthetic rates for IgE. The depressed IgE synthesis was associated with a significantly prolonged IgE half-life. Patients with Hodgkin's disease had significantly increased serum IgE concentrations. One of three patients studied had a high serum IgE concentration and synthetic rate of IgE. The two other patients had normal serum IgE concentrations associated with normal synthetic rates. Finally patients with protein-losing enteropathy or familial hypercatabolic hypoproteinemia had normal IgE concentrations associated with normal IgE metabolic parameters. In these cases, the disorder in the catabolic rate was not severe enough to affect the total amount of circulating IgE because IgE normally has a very high fractional catabolic rate. In general, IgE levels in a variety of disease states were correlated with IgE synthetic rates and abnormalities in the catabolic rate of IgE in disease did not exert an important effect on IgE concentration.


Urinary excretion of hydroxylysine and its glycosides as an index of collagen degradation. Urimary excretion of hydroxyprolin (Hyp) is one index of total collagen degradation, from all sources. Since some of the Hyp released from collagen may be further metabolized before it is excreted, other markers are necessary to measure collagen breakdown. Excretion of the glycosides of hydroxylysine (Hyl), glucosyl galactosyl hydroxylysine (Hy1[Gl)cGa1]), and galactosyl hydroxylysine (Hyl[Ga)]), more accurately reflects collagen metabolism since these products occur in specificratios in different tissue collagens and are themselves metabolized only to a minor degree. The ratios of total Hy1/Hyp and Hyl(GlcGal)/Hyl(Ga1) were measured in the urine of norma. subjects and of patients with Paget's disease of bone, hyperphosphatasia, and extensive thermal burns. In patients with extensive thermal burns the pattern of urinary Hy1 and its glycosides was consistent with degradation of collagen in dermis and fascia. When bone collagen degradation was dominant, the pattern of urinary metabolites reflected that source. Pagetic bone collagen has an amino acid composition similar to normal bone and Hy1(G1cGa1/Hyl(G1) of 0.396-0.743,vs. normal of 0.474+/-0.088. In untreated patients with severe Paget's disease of bone or hyperphosphatasia (urinary Hyp greater than 2.0 micronmol/mg creatinine) urinary Hyl/Hyp averaged 0.052+/-0.042 (0.042+/-0.009 in normal bone) and Hy1(G1cGa1)/Hy1(Ga1) 0.601+/-0.017 (0.47+/-0.009 in normal bone). When bone resorption was decreased sufficiently with calcitonin or disodium etidronate in these patients, both the urinary ratios of Hy1/Hyp and Hy1(G1cGa1)/Hyl(Gal) rose. In normal subjects treated with calcitonin and excreting relatively little Hyp, the ratio of Hy1/H)P approached 0.7 and Hy1(G1ycGa1)/Hy1(Ga1) approached 3.5. There increased ratios reveal the existence of a source of collagen breakdown other than skin or bone. The first subcompoent of complement, Clq, which has collagen-like sequences, relatively high amounts of Hy1, and most of the glycosylated Hy1 as Hy1(G1cGa1), could be the source of these metabolites.


Evidence for validity of the differential personality inventory. In order to check the concurrent validity of the Differential Personality Inventory (DPI), the authors gave the test to 60 psychiatric patients (25 men, 35 women), who also were observed and rated on Lorr and Vestre's Psychotic Inpatient Profile. Patients also took Whitaker's Index of Schizophrenic Thinking (WIST), a test that measures thought disorder. The patients received higher scores on appropriate scales of the DPI than did a normative group (N = 370) of university students. We predicted that six of the DPI scales would correlate with relevant criterion measures (Lorr-Vestre scales and WIST); in five of six instances such correlations were statistically significant. We conclude that the DPI discriminates psychotics from normals and that among psychotics it has a moderate power to discriminate among types of psychotic behavior.


The dynamics and treatment of child abuse: can primate research provide the answers? Practitioners in the mental health professions presently are faced with the tasks of understanding the dynamics involved in the problem of child abuse and devising efficacious treatment and rehabilitation programs for both the abusive parent and the abused child. The present paper reviews research done by comparative psychologists more than a decade ago that should be considered by those practitioners involved in the treatment of the family in which child abuse has occurred. Seay, Alexander and Harlow (1964) reported on a study that not only points out an etiological factor in abusive mother monkeys, but suggests treatment approaches for both the mother and her battered infant.


The volume of the uterus in normal and abnormal pregnancy. The relationship of the volume of the uterus to the bipartial diameter in single, twin, triplet and abnormal gestations was determined. The relationships are linear and the populations of single and multiple pregnancies fall into distinct groups. Correlation of the uterine volume and biparietal diameter also establishes an objective basis for diagnosing the presence of polyhydramnios ,fetal hydrocephalus, or any abnormality that produces a distortion of the normal utere volume relationship.


Retropenal abcess secondary to a ruptured retrocecal appendix diagnosed by ultrasound B scan and 67 Ga radionulide scan. Standard roentgenographic criteria for anterior-posterior (AP) displacement of the kidney may not reveal small, uniformly distributed, perirenal masses. Ultrasound, on the other hand, is an axial display and thus is quite sensitive to such changes. A case of ruptured retrocecal appendix is presented in which the intravenous pyelogram was negative. In the sonogram, however, a slight separation was seen between the renal parenchyma and the posterior abdominal muscle wall. This led to further investigation and the diagnosis of an unsuspected retrorenal abscess.


Contralateral displacement of abdominal viscera by a retroperitoneal liposarcoma: ultrasonic demonstration. Occasional reports have described contralateral displacement of the kidney, gall bladder, or liver by masses, but no ultrasonographic description of this phenomenon has been published. To our knowledge, this is the first report of contralateral displacement of all three structures by an encapsulated, well-differentiated low-grade liposarcoma. The significance to the ultrasonographer lies in the fact that he may be the first physician, as we were in this case, to have the opportunity to recognize the true anatomical relationships of all three organs and to suggest that this tumor is more likely to be benign or slowly growing rather than a highly infiltrative process.


Continuous abdominal raster scanning a technique for increasing anatomic information and reducing scan time. Continuous raster scanning can improve abdominal B scan image resolution and decrease the time required for scanning. Rapid, serial, single-sweep scans are performed at 3 to 8 mm intervals during suspended respiration. This technique markedly reduces organ distortion by respiratory and vascular motion. It is best performed with a contact scanner operating in video mode that utilizes a high resolution store oscilloscope.


Reverberations in echocardiograms.

Reverberations in echocardiograms. Different types of reverberations which may cause confusing artifacts in echocardiograms are described. Reverations between the targets may result in identifiable complex echo patterns. An echocardiogram with reverberating echoes is presented. Several experimental studies were done to explain the reverbating echoes. The reverberating echoes may be more prominently displayed than the primary echoes. This raises the possibility of obtaining useful information from the reverberating echoes.


A simple ultrasonic method for evaluating liver size. By measuring the liver area (maximum section) and plotting this against intra-abdominal area at the same level, one can calculate tolerance levels for 80% of the population (63 normal patients) with a 95% confidence. This provides a simple semiquantitative index in relating liver size of a specific patient to the normal range. Its use in 14 patients with liver disease was demonstrated.


Comparison of ultrasound pancreatic scanning and encoscopic retrograde cholangiopancreatograms: a retrospective study. In a retrospective study comparing ultrasonography and endoscopic retrograde cholangiopancreatograms (ERCP) of the pancreas we found ultrasound 93.8% accurate and ERCP 62.5% accurate in detecting pancreatic pathology. Thus, sonography provides a safe screening procedure and ERCP provides a complementary approach.


Aminoglycoside-resistant enterococci.

Aminoglycoside-resistant enterococci. Thirty-four recent clinical isolates of Streptococcus faecalis were tested for sensitivity to amoxycillin, benzylpenicillin, streptomycin, kanamycin, gentamicin, tobramycin, and amikacin. Amoxycillin was two- to four-fold more active than benzylpenicillin and all strains were inhibited by low concentrations of the penicillins. The aminoglycosides were less active against the enterococci than were the penicillins and a significant number of strains were insensitive or relatively insensitive to one or more of the aminoglycosides. Thus, eight (23%) strains showed a high level of resistance to streptomycin and kanamycin (MIC greater 5000 microng/ml) but were sensitive to gentamicin, tobramycin, and amikacin. In addition, two strains of Strep. faecalis, isolated at different hospitals from patients who had received topical gentamicin therapy, were relatively resistant to gentamicin (MIC250 to 500 microng/ml) and were less sensitive also to the other aminoglycosides. Bactericidal synergy was demonstrated by amoxycillin/aminoglycoside combinations against the enterococci, provided that the test strain of Strep. faecalis was sensitive to the aminoglycoside in the combination. An exception to this was the combination of amoxycillin plus amikacin which was not synergistic against kanamycin-resistant strains of Strep. faecalis although these organisms were sensitive to amikacin in the growth inhibition tests. The gentamicin-resistant strains showed variable responses to amoxycillin/aminoglycoside combinations in tests for bactericidal synergy and were generally less sensitive than typical strains of Strep. faecalis.


Detection of Aspergillus fumigatus precipitins: a comparison of counter immunoelectrophoresis and double diffusion. A counter immunoelectrophoresis method was compared with a double diffusion test for detecting Aspergillus fumigatus antibodies. Of 70 patients, 23 gave positive results in both tests and the remainder gave negative results. Fifteen patients with proven aspergillosis gave positive results in both tests.


Projections from cortical visual areas 17, 18, and MT onto the dorsal lateral geniculate nucleus in owl monkeys. Projections from cortical visual Areas 17, 18, and MT to the dorsal lateral geniculate nucleus of owl monkeys were revealed by the use of the autoradiographic tracing method. Restricted injections of 3H-proline into Area 17 resulted in dense columns of labeled tissue extending through all layers of the nucleus and roughly perpendicular to the layers. There was some lateral spread of the label in the interlaminar zones. Projections from Areas 18 and MT were distributed over the ventral layers of the nucleus including the magnocellular layers, layer S, and the adjoining interlaminar zones. The input from Area 18 was relatively sparse. The projections from all three cortical visual areas appear to connect homotopic locations in the geniculate and cortical representations of the visual field.


Descending pathways from the superior collicullus: an autoradiographic analysis in the rhesus monkey (Macaca mulatta). The autoradiographic tracing method has been used to identify the various descending tectofugal pathways and their targets in the rhesus monkey (Macaca mulatta). The present data reveal that the majority of descending tectofugal axons arise from collicular laminae which lie ventral to the stratum opticum (layer 3). Such descending axons can be grouped into two major bundles or tracts, i.e., the ipsilateral tectopontine-tectobulbar tract and the crossed tectospinal tract (or the predorsal bundle). There is, in addition to these two major pathways, a smaller, commissural projection. The ipsilateral pathway courses laterally and ventrocaudally to terminate within the parabigeminal nucleus, the mesencephalic reticular formation, the dorsal lateral pontine gray (in several discrete patches), the dorsal lateral wing of the nucleus reticularis tegmenti pontis, and within the nucleus reticularis pontis oralis. Other ipsilateral targets of the deep tectal layers are the cuneiform nucleus and the external nucleus of the inferior colliculus. In several experiments transported protein is also apparent within the substantia nigra. Axons which comprise the tectospinal tract, or the predorsal bundle, cross within the dorsal tegmental decussation and descend within the brainstem in a position slightly lateral to the midline. The most rostral and quite extensive target of the predorsal bundle is the nucleus reticularis tegmenti pontis. As the predorsal bundle courses caudally within the pontine tegmentum, labeled axons enter the dorsal and medial regions of both the oral and the caudal divisions of the nucleus reticularis pontis. At caudal medullary levels, the mojority of the labeled axons comprising the predorsal bundle pass ventrally to end quite profusely with the subnucleus b of the medial accessory nucleus of the inferior olivary complex. Caudal to this only a few scattered, labeled axons can be followed into the cervical spinal cord. Labeled axons also pass to the opposite, or contralateral colliculus via the tectal commissure. Such axons appear to arise and end primarily within the deeper tectal layers. In one experiment, the injection invaded the mesencephalic nucleus of the trigeminal nerve. Labeled axons were apparent within the motor nucleus, the chief sensory nucleus (quite profusely) and within the spinal or descending nucleus of the trigeminal nerve.


Role of the hippocampus in blocking and conditioned inhibition of the rabbit's nictitating membrane response. Bilateral aspiration of the dorsal hippocampus produced a disrupttion of blocking of the rabbit's nictitating membrane response in Kamin's two-stage paradigm (Experiment 1) but had no effect on the formation of a Pavlovian conditioned inhibitor (Experiment 2). The results of Experiment 1 indicated that normal animals and those with cortical lesions given conditioning to a light-plus-tone conditioned stimulus (CS) gave conditioned responses (CRs) to both the light and the tone during nonreinforced presentations of each (test phase). If, however, compound conditioning was preceded by tone acquisition, only the tone elicited a CR during testing; that is, blocking was observed. In rabbits with hippocampal lesions, however, CRs were given to both the light and the tone during testing whether or not compound conditioning was preceded by tone acquisition. The data from Experiment 2 showed that rabbits with hippocampal lesions could discriminate as well as normal rabbits and those with cortical lesions between a light (CS+) and light plus tone (CS-). In addition, when the inhibitory tone was subsequently paired with the unconditioned stimulus in retardation testing, animals in all three lesion conditions acquired the CR at the same rate. Thus, it appears that hippocampal lesions do not disrupt conditioned inhibition. The results of these experiments were taken as support for the view that the hippocampus is responsible for "tuning out" stimuli that have no adaptive value to the organism.


Comparison of procedures for measurement of IgE protein in serum and secretions. Because of conflicting reports in the literature concerning the value of various procedures for measurement of IgE in serum and secretions, we compared four different methods, the radioimmunosorbent test (RIST), the double-antibody radioimmunoassay (RIA), the paper disc immunosorbent test (PRIST), and radial immunodiffusion (RID). The standards used in the assays were tested initially in the double-antibody RIA with the use of the reference from the World Health Organization. The results showed that RID as expected was relatively insensitive and IgE was reliably measured only above approximately 1,000 international units (IU). Moreover, certain sera containing low levels of IgE by the other procedures gave distinct precipitin zones and presumably falsely high levels of IgE protein. Thus RID may yield apparently erroneous results when used as a screening procedure for measurement of IgE levels. Among the other procedures PRIST and the double-antibody RIA showed the best agreement. With serum samples RIST yielded values for IgE in the low level range higher than those given by the PRIST and double-antibody RIA. With breast milk and colostrum, values of IgE between 120 and 690 ng/ml were found by RIST, whereas IgE was not detected by double-antibody RIA and PRIST. No evidence of an inhibitor of IgE was found in breast milk, so that the apparent elevation of IgE in breast milk by the RIST is likely false. These findings confirm prior reports of spurious elevations of IgE with the RIST and indicate the usefulness of the PRIST and double-antibody RIA for the measurement of IgE in sera and secretions.


Nutritional implications of renal disease. IV. Nutritional aspects of chronic renal insufficiency in childhood. Nutritional treatment of children with renal insufficiency presents special problems related to undernutrition, i.e., insufficient caloric intake to permit normal growth, vitamin D intake, and protein needs, as well as depressed appetite. With regard to energy, it is suggested that uremia may lead to increased caloric requirements, thus exacerbating growth depression. Protein requirements, which actually may not be as important as caloric needs, have not been determined for uremic children. Another factor in growth failure in such children involves vitamin D metabolism and its role in renal osteodystrophy. Successful dietary management of these various interrelated aspects of childhood renal disease requires sensitive, knowledgeable personnel.


Controlling health care costs: strengthening the private sector's hand. The council on Wage and Price Stability recently has discovered evidence suggesting "an unackonwledged potential of the private sector to exert influence and control in the area of health care cost inflation." This article examines the limitations on private-sector cost-control efforts and suggests actions which would permit and encourage private decision makers to be more effective. In particular, private health insurers' potential role in cost control is explored, and some promising insurer strategies are adumbrated. Carefully designed coverage limitations and plan-initiated reviews to exclude nonessential care from coverage are suggested, together with wider use of fixed indemnity payments or negotiated fees and charges (instead of paying unusual and customary rates or incurred costs). Among the steps needed to permit insurers to provide this added service to cost-conscious customers is enforcement of the antitrust laws to prevent doctors' organized resistance to unwanted measures. In general, it is argued that private-sector efforts are likely to be more effective than government-sponsored controls as well as more appropriate in a pluralistic society.


Immunohistochemical detection of Fc receptor. I. Light microscopic demonstration of Fc receptor by using soluble immune complexes of peroxidase-antiperoxidase immunoglobulin G. The mouse mesenteric lymph node cells (in the cell suspension and frozen sections) were incubated in the soluble immune complexes of peroxidase-antiperoxidase immunoglobulin G. After being washed, they were reacted with diaminobenzidine tetrahydrochloride. Light microscopically brown-colored granules were observed on the cell surface of a proportion of small lymphocytes. In frozen sections, a proportion of small lymphocytes were stained dark brown on the cell surface. Characterization and control experiments suggest that the binding of peroxidase-antiperoxidase immunoglobulin G to the cell surface is mediated by Fc receptor. Peroxidase-antiperoxidase immunoglobulin G, therefore, can be used as in indicator of Fc receptor.


Immunohistochemical detection of Fc receptor. II. Electron microscopic demonstration of Fc receptor by using soluble immune complexes of ferritin-antiferritin immunoglobulin G. The mouse mesenteric lymph node cells were incubated in the soluble immune complexes of ferritin-antiferritin immunoglobulin G at 37 degrees C for 20 min. After being washed, postfixed with OsO4 and dehydrated by degraded ethanol series, the lymph node cells were observed by electron microscope. Apprroximately 15% of the cells (mainly composed of small lymphocytes) bound ferritin particles to the cell surface. The distribution pattern of the binding of ferritin particles (ferritin-antiferritin immunoglobulin G) took the form of discrete patches of irregular distribution interspaced with unlabeled portions. The electron microscopic features of ferritin particles (ferritin-antiferritin immunoglobulin G) attached to the cell surface suggest that a structure of constant conformation (Fc receptor) situated in the cell membrane takes part in the binding of ferritin-antiferritin immunoglobulin G.


Stabilization of cholesterol in myelin with digitonin: observation with polarized light. Cholesterol can be preserved in tissues dehyrated for light microscopy by incorporating digitonin into glutaraldehyde fixation. Fresh and fixed frozen sections of rat sciatic nerve exhibit strong, radially positive birefringence when viewed with polarized optics. However, tissue fixed and dehydrated with standard methods for Epon embedding shows very weak radially positive birefringence. The use of digitonin with a variety of techniques generally permits retention of optical activity in embedded nerves. Such observations suggest a preservation of lipids (presumably cholesterol) in molecular orientations necessary to produce Maltese cross patterns.


A study of the variable heavy chain (VH) region of membrane-bound Ig on human chronic leukemic lymphocytes. Lymphocytes from 20 patients with chronic lymphocytic leukemia (CLL) were studied for membrane staining by direct immunofluorescence by employing anti-F(ab')2, anti-VHI, anti-VHII, anti-VHIII subgroup-specific antisera, as well as light chain-specific antisera. Some lymphocyte preparations were also studied in indirect immunofluorescence with an antiserum raised against a fragment (VH) corresponding to the variable region of the heavy chain of a human IgG3 myeloma protein (Kup). Lymphocytes from each CLL patient demonstrated a restriction of VH subgroups expressed on the cell membrane; six were restricted to the VHI subgroup, seven to VHII, and seven to the VHIII subgroup. This restriction gave further evidence for monoclonality of the membrane-bound Ig and the leukemic cell proliferation. Antiserum to the VH fragment stained closely similar percentages of CLL lymphocytes to that obtained with anti-F(ab')2 antiserum. Furthermore, double staining revealed that the same cells were stained with anti-VH antiserum as were stained with anti-F(ab')2 antiserum, i.e., only the B lymphocytes.


Identification and genetic control of the n83 and n84 allotypes of rabbit IgM. Two additional allotypes of rabbit IgM, n83 and n84, have been identified and characterized with antisera obtained by cross immunization of rabbits with IgM. These allotypic specificities were not detected on IgG or IgA by double diffusion in agar gel and by quantitative radioprecipitin analyses. This finding implies that the specificities reside in the CH region of the mu-chain. Most of the IgM (88%) from an n83 homozygote reacted with anti-n83 Ab and most of the IgM (79%) from the net n84 homozygote reacted with anti-n84 Ab. The IgG or IgA from n83 or n84 rabbits did not precipitate to a significant extent (less than 6%) with the anti-n83 or anti-n84 Ab, respectively. That the n83 and n84 specificities are controlled at the n locus was verified by genetic analysis. Thus, four alleles are now known at the n locus, n81, n82, n83 and n84. The n locus is closely linked to the other six defined loci in the heavy chain chromosomal region and the allelic alternatives for each of the seven loci are coinherited by gene combinations called allogroups. Ten such allogroups are now defined with respect to the allele at the n locus.


T cell-mediated cytotoxicity against trinitrophenyl-modified cells: effect of glutaraldehyde treatment on the immunogenicity and antigenicity of trinitrophenyl-modified cells. Cells treated with low concentrations of glutaraldehyde for a 10-sec interval were unable to incorporate 3H-leucine into TCA precipitable protein, respond to H-2 allogeneic cells in mixed lymphocyte reactions (MLR) and cell-mediated lympholysis (CML) assays, or display capping of cell surface immunoglobulin (Ig) with a fluoresceinated anti-Ig reagent. Such cells could stimulate and specifically block H-2 allogeneic CML activity but could not stimulate an H-2 allogeneic MLR response. Trinitrobenzenesulfonic acid (TNBS) treated spleen cells were used to sensitize syngeneic splenocytes into displaying a cytotoxic effect against trinitrophenyl (TNP)-modified target cells. Treatment of the stimulator cells with glutaraldehyde immediately after modification with TNBS did not impair their immunogenic activity. Similar treatment of TNP-modified concanavalin A-stimulated lymphoblasts that were used as inhibitors in a CML cold target competition assay allowed such cells to retain their antigenicity. Cells treated with glutaraldehyde before TNP-modification, however, were not antigenic in the cold target competition assay. These data are compatible with TNBS acting on plasma membrane molecules directly to cause cells to be antigenic and immunogenic in the CML assay rather than affecting internal cellular components.


Disulfide bonding of secretory component to a single monomer subunit in human secretory IgA. The arrangement of disulfide bonds joining secretory component (SC) to the alpha chains in secretory IgA was studied by determining the molecular size of the principal fragments resulting from CNBr digestion of secretory dimeric Fc fragments from IgA (Fc)2alpha fragments). In vitro complexes formed by incubating 125I-free SC and myeloma 131I-(Fc)2alpha fragments were isolated by gel filtration and subsequently digested with cyanogen bromide. The CNBr digests of SC-(Fc)2alpha fragments were analyzed by gel filtration in 5 M guanidine. Two principal fragments were obtained, one containing a monomeric Fc fragment from IgA (Fcalpha) associated with SC (m.w. congruent to 110,000) and a second containing the second Fcalpha monomer (m.w. congruent to 50,000) from the dimeric SC-(Fc)2alpha. Similar results were obtained when secretory (Fc)2alpha fragments isolated from native secretory IgA dimer were subjected to CNBr digestion. The data indicate that SC is disulfide bonded to a single monomer subunit in secretory IgA dimer.


Enhancement of carrier-mediated transport after immunologic activation of peritoneal macrophages. Immunologically activated peritoneal macrophages from inbred mice and Hartley strain guinea pigs demonstrate a markedly greater than normal transport of 2-deoxy-D-glucose and L-leucine. The degree of nutrilite transport enhancement was greatest when animals were injected with the appropriate eliciting antigens before harvesting and also, if antigen was included in the tissue culture medium during the initial hours of in vitro culture. Enhanced hexose and amino acid uptake could also be achieved by exposure of macrophages from nonimmunized animals for 48 hr to supernatants of sensitized splenic lymphocyte cultures incubated with specific antigens. The animal systems in which this phenomenon was observed included CBA/J and C57BL/6J mice immunized with Staphylococcus aureus or sub-lethal doses of Listeria monocytogens, and the Hartley strain, albino guinea pig immunized with S. aureus or BCG. In all cases, immunization resulted in a state of delayed hypersensitivity as measured by skin testing or footpad swelling. Splenic cell supernatants contained lymphokines as detected by the presence of macrophage inhibitory factor (MIF), and by the supernatants' capacity to stimulate incorporation of 14C-glucosamine by macrophages in vitro. No increase of glucose or leucine transport by macrophages was observed in the absence of appropriate antigen stimulation in vivo or in vitro. We previously showed that a phagocytic stimulus results in a significant increase in hexose transport by normal macrophages; leucine transport by these same cells was unaltered after phagocytosis. In contrast, immunologically activated macrophages do not transport measurably more 2-deoxy-C-glucose after particle ingestion; activation or the phagocytic stimulus enhance 2-deoxy-C-glucose uptake to approximately the same extent. Analysis of nutrilite transport kinetics revealed that immunologic activation of macrophages increases the initial velocity (V1) and Vmax but does not change the Km values of hexose or amino acid transport. The kinetics of transport by the immunologically activated macrophages do not change measurably after phagocytosis. We conclude that either immunological activation or phagocytosis results in augmented 2-deoxy-D-glucose transport via identical or related mechanisms and that transport of the sugar can't be increased above that level induced by either event. The reasons why immunologic activation increases both glucose and leucine transport but phagocytosis increases only the former are not yet understood.


Isolation of porcine colostral immunoglobulins and preparation of monospecific anti-gamma, anti-alpha, and anti-mu chain antibodies using agarose-linked immunosorbents. A simple method for the simultaneous isolation of IgG2, IgG1, IgA and IgM from porcine colostral whey is described. It makes use of DEAE-cellulose chromatography followed by repeated Sepharose 6B exclusion chromatography after the removal of fat, casein and lipoproteins, Fused rocket immunoelectrophoresis is used to show the elution patterns of each immunoglobulin class. In addition, the prepation of monospecific anti-gamma, anti-alpha and anti-mu sera by means of agarose-linked immunoadsorbents is also described.


A comparative study of procedures for sheep erythrocyte-human-T-lymphocyte rosette formation. A comparative study of several published methods for sheep red blood cell-T-lymphocyte rosette formation was performed. Maximum SRBC-rosette formation occurred with AET treated SRBC in medium supplemented with 20% FCS or with untreated SRBC in 100% FCS. Prolongation of the 4 degrees C incubation period from 4 to 18 h enchanced rosette formation. Fluorescein diacetate staining significantly increased calculated percentage of rosette formation. Fluorescein diacetate staining significantly increased calculated percentage of rosette-forming lyphocytes by allowing accurate indentification of the central lymphocytes in morulas.


Organ culture studies of pemphigus antibodies. II. Ultrastructural comparison between acantholytic changes in vitro and human pemphigus lesions. The ultrastructural and light microscopic features of acantholysis produced in organ culture were compared with those of human pemphigus lesions. In both, an intraepidermal split was seen an typical suprabasal acantholytic cells were present. These cells contained small bundles of tonofilaments, usually located away from the cell periphery. Desmonsomal plaques with inserted tonofilaments frequently remained along the periphery of acantholytic cells and along the upper portion of the periphery of basal cells. The ultrastructural similarity between in vitro and in vivo lesions provides additional evidence to suggest that organ cultures may provide a valid model for studying the dynamics of pemphigus lesion formation.


Inactivated Venezuelan equine encephalomyelitis virus vaccine complexed with specific antibody: enhanced primary immune response and altered pattern of antibody class elicited. Complexes of formalinized Venezuelan equine encephalomyelitis (VEE) virus vaccine and specific IgG formed at antigen-antibody equivalence enhanced the immune responses of rhesus monkeys (Macaca mulatta). The predomonant class of antibody elicited by complexes was IgG. In contrast, lower titers of antibody and a more biphasic (IgG-IgM) response were observed after exposure of monkeys to the vaccine alone. In comparison to the response of monkeys primed with antigen, a more rapid secondary response was obtained in monkeys primed with the complexes of antigen and antibody formed at equivalence. A sustained level of protection of 88% was afforded mice 24 hr after immunization with antigen-antibody complexes; development of protection after administration of antigen required eight days to reach this level. Passive protection (80%-100%) was conferred by IgG controls for seven to eight days after immunization. This level of protection was not significantly affected by X-irradiation 24 hr prior to administration of IgG; however, protection in mice similarly irradiated prior to immunization with antigen-antibody complexes was significantly decreased. Early protection afforded by the complexes was not nonspecific (interferon) but was mediated by specific immunologic mechanisms and may be caused by an early formation of IgG.


Patterns of rickettsemia and antibody response in silvered leaf monkeys (Presbytis cristatus) after inoculation with virulent and avirulent strains of Rickettsia tsutsugamushi. Most silvered leaf monkeys inoculated with selected strains of Rickettsia tsutsugamushi developed rickettsemia. The time of onset and the duration of rickettsemia were related to the infecting strains, but these parameters could not be correlated with the virulence of the strain. The rickettsemia was not terminated by the appearance of humoral antibody, as detected by the indirect fluorescent antibody test. The specificities of the antibody responses were related to the antigens present in the infecting strains.


Neisseria meningitidis infections in Northern Norway: an epidemic in 1974-1975 due mainly to group B organisms. A severe epidemic of disease due to Neisseria meningitidis has been occurring in northern Norway since 1974. Based on bacteriological and clinical examination of cases during the 18 months between January 1, 1974 and June 30, 1975, an overall attack rate of 26.3 cases per 100,000 population per year has been calculated. The attack rates were highest in 1975 (total, 37.4 cases per 100,000 population; Troms County, 54.5) and in children younger than four years (total, 173.6; Troms County, 328.9). The epidemic shows a total case-fatality ratio of 14.1%. The highest case fatality ratio was recorded for patients older than 20 years (25.0%). Among the youngest children, cases were distinctly more frequent in males than females; in older children and adults, male patients had a clearly higher case-fatility ratio than female patients. Of the three serogroups of N. meningitidis, group B caused more cases of infection that groups A and C together. In addition, the causative meningococcal strains were almost uniformly resistant to high concentrations of sulfonamide. Both of these traits appear to have developed in northern Norway at or shortly before the time when the epidemic began.


Pseudomonas species bacteremia caused by contaminated normal human serum albumin. In May and June 1973, 11 patients on the surgical service at the University of Maryland Hospital had bacteremia caused by Pseudomonas species. Seven of the isolates recovered from blood cultures had the same antibiogram (sensitive only to chloramphenicol and tetracycline). Ten of the 11 patients were given 25% normal serum albumin (human) shortly before the onset of symptoms. In contrast, only two of seven patients with bacteremia due to Psuedomonas aeruginosa in May and June (P =0.013) and only nine of 20 patients located in surgical special care units during these months (P =0.014) were given this product. When cultured, the albumin in one of 54 previously unopened vials from the implicated lot yielded Pseudomonas cepacia sensitive only to chloramphenicol, tetracycline, and nalidixic acid. Subsequent investigation showed that five more patients in four other hospitals had symptoms of bacteremia shortly after the infusion of different lots of albumin from the same manufacturer, and in four cases P. cepacia was cultured from the suspect albumin. Since sterility testing by manufacturers may not detect low-frequency contamination, surveillance of nosocomial infections, investigation of unusual disease clusters, and prompt reporting of suspect reactions are essential in the control of such outbreaks.


Pathogenesis of canine bordetellosis.

Pathogenesis of canine bordetellosis. Bordetella bronchiseptica produced tracheobronchitis when administered in aerosol to specific pathogen-free dogs. Clinical signs appeared to be directly related to numbers of bacteria in the trachea. Electron microscopic examination revealed that each bacterium was close to one or more tracheal cilia and that a fibrillar material was radiating from the bacterial cell wall. B. bronchiseptica required 14 weeks to be cleared from the tracheas of infected dogs; in contrast, other organisms commonly isolated from the respiratory tracts of dogs were cleared within one to three days. Strains of high and low in vitro passage and strains representing three different morphotypes were of equal pathogenicity. Local immunity was observed after infection and appeared to be of primary importance in recovery from the infection. Presumably this response involves prevention of bacterial attachment and reattachment to cilia.


Enhancement of nonspecific immunity to bacterial infection by cord factor (6,6'-trehalose dimycolate). The capacity of mycobacteria to enhance the nonspecific resistance of mice to bacterial infections was related to the presence of cord factor. After enzymatic digestion and delipidation, cell walls lost their immunopotentiating activity, which was restored, however, by the addition of cord factor. Cord factor administered alone protected mice effectively against virulent organisms unrelated to mycobacteria. Moreover, although this glycolipid is usually injected as a solution in mineral oil, our data show its effectiveness when administered in metabolizable peanut oil or even as a suspension in water. Since cord factor is a chemically well-defined molecule (6,6'-trehalose dimycolate), this glycolipid should constitute a new and useful tool for the investigation of nonspecific immunity to infection.


Pharmacokinetics of methicillin in patients with cystic fibrosis. The disposition of methicillin in normal subjects and in subjects with cystic fibrosis (CF) was studied after administration of single intravenous doses of 15 mg/kg. The area under the serum concentration vs. time curve for CF patients was, on the average, only 75% of that found for normal subjects. The low concentrations in serum were caused by more rapid urinary excretion of the antibiotic, with rates of renal clearance averaging 425 ml/min per 1.73 m2 in the patients with CF and 362 ml/min per 1.73 m2 in the normal subjects. No differences were found in volumes of distribution and metabolic clearance rates of methicillin or in rates of creatinine clearance between the two groups of subjects. These data support previous findings with dicloxacillin which show that patients with CF exhibit unusually rapid, active tubular secretion of certain penicillins that may necessitate use of larger doses of these drugs in treatment of infections.


Administration of human interferon to rabies virus-infected monkeys after exposure. The treatment of rabies-infected cynomolgus monkeys (Macaca fascicularis) with human interferon after exposure was studied. The monkeys were infected with rabies street virus by the intramuscular route; larger than or equal to 24 hr after infection, human interferon was administered intramuscularly or by lumbar spinal injection into the cerebrospinal fluid. Whereas 90% of the infected untreated monkeys died, 40%-80% of animals treated with interferon survived. No or only low levels of neutralizing antibody to rabies virus were found in the sera of monkeys that survived after a single intramuscular injection of human interferon. Distinctly higher antibody titers, however, were detected in the sera of surviving monkeys that had been given six consecutive intralumbar doses of interferon beginning three days after infections. Therefore, we conclude that in these monkeys rabies virus propagated until it spread into the central nervous system, where further viral replication was inhibited by the human interferon administered by the intralumbar route.


Effects of a plasticizer leached from polyvinyl chloride on the subhuman primate: a consequence of chronic transfusion therapy. The effects of di-2-ethylhexyl phthalate (DEHP) on hepatic function and histology were evaluated in the rhesus monkey undergoing chronic transfusion. DEHP was solubilized as a result of leaching by platelets or plasma stored in polyvinyl chloride (PVC) blood bags. The average cumulative amount of DEHP infused in 1 year was 69.3 mg., or 21.3 mg./kg. Comparable amounts of DEHP are received by patients who undergo chronic transfusions with whole blood, packed cells, platelets, and plasma in the same time period. Four of the seven PVC-transfused monkeys had abnormal 99mTc liver-spleen scan ratios, four of the seven demonstrated abnormal BSP clearance, and six of the seven had abnormal liver histopathology upon completion of transfusion therapy. These findings persisted in a majority of the monkeys throughout the follow-up period. DEHP persisted in the tissues for up to 14 months after transfusion. The nontransfused monkeys and those transfused with platelets processed in polyethylene containers were normal.


Immunochemical studies on the nature of the serum component (SAA) related to secondary amyloidosis. The relative occurrence of the amyloid-related serum protein SAA in various disease states and in healthy subjects has been compared by both solid phase radioimmunoassay ((RIA) and immunodiffusion techniques which employ antibodies to purified amyloid fibril proten AA of hemogeneous size and charge. SAA levels were elevated above normal in certain categories of neoplastic, inflammatory, and infectious diseases as well as in secondary amyloidosis. The lowest median value, 8 ng./ml., was observed for approximately 150 normal sera, with no age-related increase in subjects ranging in age from 16 to 70 years. The results are consistent with several recent observations that SAA is normal acute phase reactant, and hence the RIA for SAA has no prognostic or diagnostic significance for secondary amyloidosis. The sensitivity of RIA for the detection of SAA is lower than would be expected when AA cross-reactivity values for sera are correlated with their reaction with anti-AA antibodies in immunodiffusion. This observation, along with others reported elsewhere suggests that those determinants which cross-react with anti-AA antibodies are relatively hidden in native SAA. Myeloma sera were less reactive than other groups of pathologic sera in immunodiffusion, although they were similar to other patients' sera when analyzed by RIA. Antibodies to highly purified AA were also used to investigate the structue of SAA by a double-antibody immunoprecipitation method. Precipitated SAA  was partially dissociated during sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis to a 12,500 molecular weight moiety designated, SAAL. Multiple radiolabeled species of molecular weight intermidiate to SAA and SAAL were also detected and appeared to represent imcompletely dissociated SAA. The results suggest the SAA is an aggregate of several SAAL chains.


Preparation of type-specific anti-M streptococcal sera by ummunization with acid-hydrolyzed cells. The present study describes a method which simplifies the preparation of type-specific anti-M streptococcal sera. The procedure is based on vaccination with acid-hydrolyzed cells. The method makes possible the preparation of antisera from M strains which are problematic by conventional vaccination and is very useful for preparing all anti-M sera. The antisera obtained by the above method are devoid or almost devoid of cross-reactive antibodies.


Properties of IgA myeloma proteins isolated rom sera of patients with the hyperviscosity syndrome. Sera, purified myeloma proteins, and urinary proteins obtained from eight patients with igA multiple myeloma were studied by physical-chemical and immunochemical methods. In six patients whose serum viscosity was increased, the sedimentation constants of the principal component of myeloma proteins ranged from 9.1 to 10.2 S. In two patients with nearly normal serum viscosity, the sedimentation constants of these proteins were 6.2 and 7.2 S. IGA-albumin complexes were detected in most of the sera, but invarying amounts; no complexes of ig with amylase, secretory component, or alpha(1)-antitrypsin were observed. Studies on isolated myeloma proteins revealed that all igA proteisn from sera with increased viscosity represented true polymers, linked by disulfide bonds, rather than noncovalently associated aggregates; J chain was detecable by both alkaline-urea disc electrophoresis and immunoelectrophoresis with a monospecific anti-J chain serum. Increased serum viscosity was not related to the igA subclass, L chain type, or the carbohydrate compositions of individual igA myeloma proteins. The urine of five patients contained free light chains corresponding in type to the light chain of the particular igA myeloma protein. However, free J chain was not detected. The immunoelectrophoretic analysis for the presence of J chain in sera of myeloma patients may be used for early and simple detection of polymeric forms of myeloma proteins.


Treatment of acute suppurative otitis media. The effect of myringotomy versus Penicillin therapy in the treatment of patients with acute suppurative otitis media was compared. A third group of patients treated with both myringotomy and Penicillin was included in the investigation. Ten days after institution of the treatment there were significantly more days with secretion/bulging drum in the myringotomy group than in the two other groups. At this time Penicillin therapy was started also in those patients belonging to the myringotomy group. With this reservation the occurrence of such late complications as secretory otitis media and relapses was the same, independent of treatment. No cases of mastoiditis were observed.


Thyrotrophin releasing hormone-induced growth hormone and prolactin release: physiological studies in intact rats and in hypophysectomized rats bearing an ectopic pituitary gland. To determine how the sensitivity of the ectopic anterior pituitary gland to the GH-releasing effect of thyrotrophin releasing hormone (TRH) might be affected by the time lapse from transplantation, TRH (0-15 and 0-6 microng) was injected i.v. into hypophysectomized (hypox)-transplanted rats under urethane anaesthesia 1,3,8,15,30 and 60 days after transplantation, and plasma samples were taken 5 and 10 min later. Baseline GH values gradually decreased with time from about 16-0 ng/ml (1 day) to about 3-0 ng/ml (30 and 60 days). The TRH-induced GH release was absent 1 day after transplantation, present only with the higher TRH dose 3 and 8 days after transplantation, and clearly elicitable, also with the lower TRH dose (0-15 microng), from 15 up to 60 days. Determination of plasma prolactin concentrations showed a decline from about 85-0 ng/ml (1 day) to about 32-0 ng/ml (8 days); subsequently (15-60 days) prolactin values stabilized. Plasma prolactin levels increased 15 and 60 days after transplantation only when a dose of 0-6 microng TRH was given. In intact weight-matched rats, TRH induced a GH response only at the dose of 1-2 microng while a short-lived but clear-cut prolactin response could be obtained even with the 0-3 microng dose. The present results indicate that: (1) disconnexion between the central nervous system and the anterior pituitary gland greatly enhances GH responsiveness while blunting prolactin responsiveness to TRH; (2) the sensitivity of the anterior pituitary gland to the GH-releasing effect of TRH increases with time from transplantation; (3) TRH is a more effective prolactin- than GH-releaser on the pituitary gland in situ.


Light and electron microscopic studies of thyrotrophin releasing hormone-induced growth hormone and prolactin release in hypophysectomized rats bearing an ectopic pituitary gland. A light microscopic and ultrastructural study of anterior pituitary glands transplanted under the kidney capsule of hypophysectomized rats was performed, in basal conditions and after stimulation with thyrotrophin releasing hormone, at various intervals (24 h-2 months) after transplantation. Confirming previous biochemical findings, the results suggest that with time, somatotrophs acquire sensitivity to thyrotrophin releasing hormone and respond with increased exocytosis at doses that were found ineffective in pituitary glands in situ. Thyrotrophin releasing hormone stimulation did not seem to influence the morphology of prolactin cells, which were already highly active under basal conditions at all time intervals.


Evidence for dopaminergic control of thyrotrophin secretion in the rat. The effects of two dopamine agonists (apomorphine and bromocriptine) and a dopamine antagonist (pimozide) on cold- or thyrotrophin releasing hormone (TRH)-induced TSH secretion were studied in normal male rats. Apomorphine given in various doses (0-5-10 mg/kg body wt) 10 min before exposure to cold significantly depressed TSH secretion. Large doses of bromocriptine (5-10 mg/kg body wt) given 1 h before exposure to cold, also blocked this response whereas a smaller dose (2-5 mg/kg body wt) given 30 min, 1, 3 or 6 h before cold exposure or repeated doses (0-1-2-5 mg/kg body wt) for 3 days did not modify cold-induced TSH secretion. Pimozide given in various doses (-25-2-5 mg/kg body wt) 1 h before exposure to cold did not alter the cold response, but 2-5 mg/kg reversed the inhibition caused by apomorphine or bromocriptine. None of these drugs affected TRH-induced TSH secretion. These results suggest that there are no dopaminergic receptors on the pituitary thyrotrophs, but that dopamine might be an inhibitory transmitter in the brain involved in the regulation of TSH secretion in the rat.


Early increases in ribonucleic acid polymerase activities of dimethylbenzanthracene-induced mammary tumour nuclei in response to oestradiol-17beta and tamoxifen. Studies on the mode of action of tamoxifen have shown that this compound ultimately causes regression of mammary tumours induced in female rats by 7,12-dimethylbenz(a)-anthracene, but induces preliminary effects similar to those produced by oestradiol-17beta. Following a single intravenous injection of either substance, a sequence of events was observed which included depletion of cytoplasmic receptor, a concomitant increase in nuclear receptor and a subsequent replenishment of cytoplasmic receptor. Tamoxifen and oestradiol-17beta induced a transient increase in RNA polymerase B activity, followed by increases in RNA polymerase A and, again, RNA polymerase B activity. Tamoxifen, unlike oestradiol-17beta, could not maintain replenishment of cytoplasmic receptor, the increase in RNA polymerase A activity or the secondary rise in RNA polymerase B activity. The basic anti-oestrogenic properties of tamoxifen may be implicit in its inability to maintain oestrogen stimulation, and may be linked to its retention time within the nuclei.


The eversion and differentiation of Drosophila melanogaster leg and wing imaginal discs cultured in vitro with an optimal concentration of beta-ecdysone. The stages of the eversion and differentiation of prothoracic leg and wing discs of Drosophila melanogaster, in Shields and Sang's medium 3, are described. The range of specific imaginal structures produced, including patterns of sensilla trichodea and sensilla campaniformia, are noted, and the relationship of these structures to those differentiated in situ is discussed.


Active ion transport and beta-acdysone induced differentiation of Drosophila melanogaster imaginal discs cultured in vitro. The theory that beta-ecdysone initiates developmental changes during insect metamorphosis by causing an increase in intranuclear levels of potassium, together with a concomitant decrease in sodium levels, has been investigated by two methods. First, imaginal discs from late third instar larvae have been cultured with 0-2 microng/ml of beta-ecdysone together with inhibitors of active ion transport. Non-specific inhibitors, which may have general effects on sulphydryl groups, such as iodoacetic acid, N-ethylmaleimide ethacrinic acid and furosemide, inhibit both eversion and differentiation at concentrations of from 10(-3) M to 2 X 10(-3) M. Ouabain, the only specific inhibitor of the active transport of Na+ and K+ across membranes, had no effect on development even at a concentration of 10(-2) M. Second, a medium containing raised levels of K+, and reduced concentrations of Na+, neither initiated disc development in the absence of beta-ecdysone, nor stimulated development induced by suboptimal levels (0-02 microng/ml) of beta-ecdysone, either in the presence or absence of ouabain. These results suggest that beta-ecdysone induced morphogenesis is not dependent upon Na+ and K+ concentrations, or on the activity of an ouabain-sensitive ion pump.


Sex determination in germ line chimeras of Drosophila melanogaster. Of 55 flies developing from blastoderms which had received male or female pole cell transplants, 15 (7 females and 8 males) wer shown by progeny testing to be germ line chimeras. Since donor and host pole cells were genetically marked with contrasting X- or Y-linked alleles, the progeny testing scheme enabled the genotypic sex of the donor component undergoing gametogenesis to be identified as either the same as ('homosexual' chimeras) or opposite ('heterosexual' chimeras) that of the host. All seven of the female chimeras were identified as 'homosexual' chimeras carrying only chromosomally female XYX donor and XX host germ cells. Similarly, all eight males were shown to be 'homosexual' chimeras with chromosomally male XY donor and XY host germ cells. The chromosomal sex of the donor component undergoing gametogenesis was in every case the same as the phenotypic sex of the host. Since there is an equal probability of constructing either a 'homosexual' or a 'heterosexual' chimera during pole cell transplantation, the ability of pole cells to differentiate functional gametes in hosts of the opposite sex was tested 50% of the time even if sex reversal of these donor pole cells could not be demonstrated. Thus the absence of 'heterosexual' chimerism strongly supports the interpretation that the phenotypic sex of a germ cell in Drosophila is determined entirely by its own chromosome constitution, not by that of the gonadal mesoderm.


The monoclonality of human B-cell lymphomas. Human tissues involved with lymphoma have been examined in frozen sections for immunoglobulin-bearing cells by a technique involving double-label immunofluorescence with mixed anti-kappa and anti-lambda antibodies. F (ab')2 fragments of purified antibodies were employed to avoid any binding via Fc receptors. B cell lymphomas were shown to be composed of monoclonal populations of Ig bearing cells, whereas normal or reactive lymphoid follicles contained a mosaic of Ig-bearing cells derived from multiple clones. Nodules of lymphoma were often surrounded by normal polyclonal B cell populations. We anticipates that the approach described here will be useful in the diagnosis of lymphoma, differentiating it from reactive lymphoid hyperplasia by the demostration of monoclonality. In addition, it should provide a sensitive and reliable tool for investigating the immunobiology of human lymphoma.


Subversion of host defense mechanisms by malignant tumors: an established tumor as a privileged site for bacterial growth. Mice carrying any one of three murine tumors in their right hind foot pad were incapable of eliminating an inoculum of the bacterial parasite Listeria monocytogenes from the progressive tumor. In contrast, they were as capable as control mice in efficiently eliminating the organism from their contralateral tumor-free foot pad, and from their lymph nodes and livers. The results serve to show, therefore, that conditions within an established tumor are not only antagonistic to the expression of concomitant anti-tumor immunity, but that they are also antagonistic to the expression of T-cell-mediated anti-bacterial immunity. The possibility was discussed that the tumor contains factors that act pharmacologically to locally suppress the function of sensitized T cells and activated macrophages.


Experience of a physician-nurse practitioner team in care of patients in skilled nursing facilities. The use of a physician-nurse practitioner team is advocated as an approach to delivering better health care to patients in skilled nursing facilities. The application of this approach in a young community with an inadequate supply of primary physicians and 596 extended care beds is discussed. Patients derive benefit from more comprehensive health care delivered with greater attention to individual needs. Staffs of skilled nursing facilities enjoy improved communication with the medical team and better compliance with legal requirements. The team physician is able to use his time more effectively and provide medical supervision for a greater number of patients by sharing responsibilities with a nurse practitioner. Paper compliance, adherence to agency regulations, quality assurance, and payment are some of the problems encountered.


Accumulation and degradation of three major yolk proteins in Drosophila melanogaster. In contrast to previous findings, three major yolk proteins have been identified in the oocytes and eggs of Drosophila melanogaster. They are also present as major proteins in the haemolymph of mature females and in trace amounts in the haemolymph of young females; the male haemolymph lacked all three proteins. Female fat body contained the three proteins and, surprisingly, trace amounts were also present in the male fat body. The accumulation and degradation of the three yolk proteins by oocytes and embryos was asynchronous suggesting that independent controls may exist.


A simple quantitative assay for bacterial motility. It is argued that the average motility of bacterial populations should be identified with a diffusivity parameter. A simple capillary assay for quantifying this parameter is described, and some results obtained by using this procedure are presented. It is concluded that the assay combine speed and simplicity of operation with sufficient accuracy to make it a valuable tool in the assessment of motility.


The isolation and characterization of lipopolysaccharide-defective mutants of Pseudomonas aeruginosa PAC1. Mutants with defective lipopolysaccharides (LPSs) were isolated from Pseudomonas aeruginosa PACIR (Habs serogroup 3) by selection for resistance to aeruginocin from P. aeruginosa PI6 Carbenicillin-sensitive mutants were isolated from P. aeruginosa PACI but not all had defective LPSs. Rough colonial morphology and resistance to bacteriophage II9X appeared to be independent of LPS composition. The LPSs from five mutants were analysed and compared with that of the parent strain. Separation of partially-degraded polysaccharides from LPS from PACI on Sephadex G75 yielded two different high molecular weight fractions and a phosphorylated low molecular weight fraction (L). The mutant LPSs lacked most or all of the high molecular weight fractions but retained some low molecular weight material. That from PACI and two of the mutants was separated by elution from Biogel P6 into two fractions. One, L2, was the core polysaccharide while the other, LI, contained short antigenic side-chains attached to the core like the semi-rough (SR) LPSs of the Enterobacteriaceae. The two mutants which gave the LI fraction with Habs 3 and PACI antisera as did the parent strain. The other three mutants were unreactive and their LPSs contained core components only. One appeared to have a complete core while the other two lacked rhamnose and rhammose plus glucose respectively. Thus there may be four types of LPS in PACI: one contains unsubstituted core polysaccharide and yields L2 on acid hydrolysis, another has short antigenic side-chains of the SR type and yields the LI fraction, while the two high molecular weight fractions are derived from core polysaccharides with different side-chains.


Biochemical characterization of infantile gastroenteritis virus (IGV). Enzymic and biophysical studies with purified infantile gastroenteritis virus (IGV) nucleic acid indicated that the virion contained a double-stranded RNA genome of approx. 14 x 10(6) daltons which could be separated by gel electrophoresis into eight bands of RNA which were comprised of 15 RNA species. Two major virus proteins, VP2 (mol. wt. = 135,000) and VP8 (mol. wt. = 40,000), which composed about 85% of the total virion protein, were detected in IGV particles by polyacrylamide gel electrophoresis. Eight additional minor proteins were also resolved.


The role of defective interfering particles in persistent infection of Vero cells by measles virus. Persistent infections by measles virus were rapidly established in the majority of Vero cells when monolayers were infected with virus stocks that had been passed three to five times from an undiluted inoculum. These virus stocks had low infectivity titres but normal haemagglutinin titres and were able to cause interference. The ability of such virus stocks to establish persistent infections seems to be due to the presence of defective interfering particles rather than of virus mutants. Measles virus released from a persistently infected Vero cell line at the 93rd passage had properties similar to the undiluted passage virus that generated persistent infections.


An experimental cerebral missile injury model in primates. An experimental model of cerebal missile injury in rhesus monkey is described. The main objective was to create a "clean" wound devoid of bleeding from major vessels and complications due to bone fragments. There was a correlation between the wounding energy and the physiological signs, although we underestimated the actual energy level. After the right parietooccipital to right frontal injury, there was bradycardia, changes in blood pressure, and, in high-energy wounds, a marked alternation in resperation. This suggests that the missile's energy produces direct brain-stem damage, the extent of which can be related to the wounding energy.


Physiological consequences of experimental cerebral missile injury and use of data analysis to predict survival. The authors describe cerebrovascular and cerebral metabolic changes in monkeys, subjected to cerebral missile injury. After injury with BB pellet at 90 m/sec, there is a rapid rise in intracranial pressure (ICP), which reaches a peak 2 to 5 minutes posttrauma, and then falls to about 20 to 30 mm Hg. This, with a fall in mean blood pressure (MBP), results in a 50% reduction in cerebral perfusion pressure (CPP), Cerebral blood flow (CBF) is also reduced, although acutely there is no close relationship with (CPP). Cerebrovascular resistance falls initially and then at 30 minutes rises to very high values. Cerebral metabolic rates (CMR's) for oxygen fall after injury and remain low for the rest of the animal's life; CMR's for lactate rise immediately after injury and persists for 5 hours, then fall. After injury with a faster missile (180 m/sec), the ICP rises higher and faster, and the peak is shorter. The CCP is reduced in this injury to approximately 30 mm Hg, and only one animal survived more than 1 hour. With the conventional forms of data analysis, the length of survival after injury correlates well with MBP, ICP, and CBF, but separately they were completely unsatisfactory for prediction of an individuals prognosis. With the technique of multiple linear regression analysis, the survival of individual animals could be predicted with great accuracy. This is possible also when two postinjury parameters,CBF and MBP, are used.


Radiographic morphometry and osteopenia in spinal osteoporosis. Measurements of bone mineral content and total-body calcium, normalized and expressed as ratios, were compared with radiographic morphometry in 45 women who had spinal osteoporosis. The radiographic indices examined included the femoral score, the femoral trabecular pattern, the biconcavity and metacarpal indices, and the total peripheral score. Both ratios and all the radiographic indices except the femoral trabecular pattern were found to be related to the number of dorsal spine fractures. The relationships observed support the following conclusions: (A) the femoral score and the metacarpal index are related to the degree of osteopenia; (B) the biconcavity index reflects the extraskeletal factors that are pathogenic in spinal osteoporosis; (C) a reduced femoral trabecular pattern index is associated with spinal osteoporosis, although this measurement is not related to the degree of osteopenia; and (D) it may be imprudent to diagnose osteoporosis from the presence of lumbar compression fractures.


Whole-body retention of radioxenon.

Whole-body retention of radioxenon. The total-body retention of 127Xe, and its variation with time following short, nonequilibrium periods of inhalation, were measured in vivo so as to refine dosimetry calculations. Radioactivity in the chest region and in the recirculating xenon-air mixture was measured continuously during re-breathing of the gas mixture and in the first 5 min of the immediate gas-washout period using a scintillation camera and shielded NaI detector, respectively. Subjects were then counted in a whole-body counter at varying time intervals until background levels were reached, usually in 72 hr. Five components of Xe clearance from the entire body were measured; they had biologic half-times of 21.7+/-12.4 sec, 3.05+/-1.72 min, 0.04+/-0.11 hr, and 2.71+/-0.87 hr, and a long-term component varied between 7.59 and 17.04 hr. The half-time of the last component correlated highly with the percent of total-body fat. After 10-min inhalations of the xenon-air mixture, approximately one-third of the total xenon was transferred to the body tissues, extrapolated back to the start of gas washout. Of this amount, 13% was associated with the slowest component of clearance.


Gallium-68 labeled red cells and platelets: new agents for positron tomography. Red cells and platelets are labeled with Ga-68 by preparing the tga-68-hydroxy-quinoline (oxide) complex and mixing the complex with the separated cellular components. The complex is prepared by first breaking the Ga-68 EDTA chelate with concentrated hydrochloric acid, adsorption of the galluim on a strong anionic ion-exchange column, and elution of the gallium activity with distilled water. Oxine is added, the oxine complex extracted into chloroform, and--following evaporation and dissolution--the solution is added to either red cells or platelets that have been centrifuged from whole blood, washed, and suspended in normal saline. After 15 min of incubation , labeling efficiencies of greater than 90% and yields of 7 mCi are obtained from 18 mCi of Ga-68 EDTA eluted from the generator. After injection of the gallium-labeled red cells in dogs, excellent blood pool images were obtained by positron tomography. The distribution corresponded to that of 11CO-labeled red cells. When labeled platelets were injected into dogs with experimentally induced intimal injury of on carotid artery, the area of injury was clearly visualized 40 min after tracer injection.


Clinical evaluation of some phosphorus bone-imaging agents: concise communication. Four different Tc-99m labeled phosphorus-based bone-imaging agents were compared by a scan-evaluation technique in which the skeletal uptake was visually assessed at selected sites. A detailed statisticalanalysis of scans, by each of three independent evaluators, on a total of 140 different patients showed that two of the agents (based on pyro- and on trimetaphosphate) were superior. The statistical analysis showed good agreement between the investigators and consistency in thier repeat evaluations.


Effect of type of dietary fat and cholesterol on cholesterol absorption rate in squirrel monkeys. To explore the effect of the type of dietary fat and the level of cholesterol on the rate of cholesterol absorption, 13 different male squirrel monkeys were used for 39 different tests. The plasma isotope ratio technique of Zilversmit, which involved the injection of 3H-cholesterol and gastric intubation of 14C-cholesterol, was shown to give reproducible results which compared well with those based on a method involving labeled beta-sitosterol. The percentages of ingested cholesterol that were absorbed showed considerable variation among individuals, but were relatively constant in the same animal. Safflower oil, a highly unsaturated fat, promoted a higher rate of cholesterol absorption than butter, regardless of the level of dietary cholesterol. The rate of cholesterol absorption was a constant percentage of that fed regardless of the absolute level of cholesterol in the habitual diet or the test meal (up to 218 mg or 5+ mg/kcal). Even at low levels of dietary cholesterol the percent absorption is greater in squirrel monkeys than in man.


Long-term effect of low dietary calcium:phosphate ratio on the skeleton of Cebus albifrons monkeys. Wildcaught cinnamon ringtail monkeys, Cebus albifrons, were fed diets with Ca:P ratios of 1:4 1:2.1 1:0,4, and 1:0.5 for 3 to 88 months. Monkeys fed the diet with Ca:P ratios of 1:4 and 1:21 C ratios similar to that of human diets) had minor microscopic changes suggestive of osteoporosis when compared to other species of animals. The changes were not detected by conventional or magnification radiography or by 125I photon absorptiometry. These findings are in in striking contrast to studies in other animals where similar diets resulted in significant bone resorption within 6 weeks to 6 months. This study suggests that the non-human primate may be a more appropriate animal model for the investigation of nutritional osteopenia in man in whom bone resorption appears to be a slowly progressive process. In view of our findings, studies using lower animal species must be re-evaluated with respect to the hypothesis that high dietary phosphate is a significant etiologic factor in senile osteoporosis in man.


Genetic heterogeneity within the chondroitinsulphaturias. The approach, identification of clinical phenotype followed by lysosomal enzyme assays in cell culture, used in the classification of the genetic mucopolysaccharidoses I-VI has been applied to the chondroitinsulphaturias. There was evidence of heterogeneity in the first 9 patients reported.


GM1 gangliosidosis type 1 in twins.

GM1 gangliosidosis type 1 in twins. This report describes 7-month-old monozygotic twin female infants with GM1 gangliosidosis type I. In addition to the usual clinical and biochemical abnormalities generalized intracutaneous telangiectasis were present in both infants.


Heterozygote detection in angiokeratoma corporis diffusum (Anderson-Fabry disease). Studies on plasma, leucocytes, and hair follicles. Heterozygote detection for angiokeratoma corporis diffusum (Anderson-Fabry disease, ACD), an X-linked disorder of glycosphingolipid metabolism was examined using alpha-galactosidase activity, an alpha-galactosidase/beta-galactosidase activity ratios (alpha/beta ratio) in leucocytes, plasma, and hair follicles; For leucocytes, 22 obligate heterozygotes, 25 suspected heterozygotes, and 47 control subjects were studied, while for plasma, the groups were 17 obligate heterozygotes and 35 controls. The alpha/beta ratio in plasma and leucocytes was clearly a better discriminator between obligate heterozygotes and controls than alpha-galactosidase activity alone, but still failed to detect 3 obligates with leucocytes and 2 with plasma. Discrimination was not improved by joint use of plasma and leucocyte alpha/beta ratios, but was improved by measurement of hair-follicle alpha/beta ratios. The interdecile range of log (alpha-galactosidase/beta-galactosidase activity) in 20 hair follicles from each of 4 obligate and 7 suspected heterozygotes was clearly different from 11 control subjects. Accordingly, for rapid screening for carriers of ACD, we recommend use of leucocyte or plasma alpha/beta ratios which should detect greater than 85% of heterozygotes. When results are equivocal, and ancillary information suggests heterozygous status, the more time-consuming measurement of hair-follicle alpha/beta ratios is a useful additional test.


Thyroid function at birth in infants of 30 to 45 weeks' gestation. Serum T4 was measured in cord blood of 2,683 infants of gestational ages 30 to 45 weeks and birth weights of 1,460 to 5,250 gm. In addition, serum TBG was measured in 180, and serum TSH in 125 of these infants. Mean serum T4 increased with gestational age from 9.4 microng/dl at 30 weeks to 11.7 at 45 weeks. TBG varied from 3.1 to 11.4 mg/dl but showed no correlation with gestational age. There was a positive correlation between T4 and TBG. Serum TSH levels showed a significant negative correlation with gestational age, decreasing from 15 micronU/ml at 30 weeks to 7 micronU/ml at 45 weeks, while the serum T4/TSH ratio increased from 0.6 to 1.6. The results indicate that cord, T4 varies mainly with TBG, and to a lesser extent with gestational age.


Serologic test for antibody to Sarcocystis in cattle. Soluble antigen was prepared from Sarcocystis zoites obtained from heart muscle of a bovine inoculated with sporocysts from canine feces and killed 120 days after infection. The antigen was used in an indirect hemagglutination (IHA) test and an agar gel diffusion test to detect antibody to Sarcocystis in experimentally infected cattle. IHA serum titers began to rise 30 to 45 days after infection and reached levels up to 1:39,000 90 days after infection. Sera collected under field conditions from 21 dairy cows had IHA titers ranging from 1:54 to 1:486. Since all cows appeared in good health, titers of 1:486 or less can probably be considered nonsignificant with regard to diagnosis of clinical disease. No positive Sarcocystis IHA titers were obtained with human sera previously found to be IHA positive for toxoplasma, indicating a lack of cross reactivity between antigens. Precipitins in the agar gel diffusion test appeared 30 days postinoculation and became very pronounced at 65 to 90 days.


The exoerythrocytic stages of Plasmodium fragile in Macaca mulatta monkeys. Exoerythrocytic (EE) schizonts of Plasmodium fragile were found in liver tissue acquired 7, 8, 9, and 10 days following inoculation of Macaca mulatta monkeys with sporozoites from Anopheles balabacensis balabacensis mosquitoes. There was little to distinguish the EE bodies of P. fragile from EE bodies of many of the other species of primate malaria.


The reactions of Schistosoma mansoni miracidia to light. The reactions of Schistosoma mansoni miracidia to artificial light were studied by direct observations, choice-chambers and a photographic technique, at room temperature and at 15 C. The miracidia responded positively to directed illumination, but did not aggregate in a lighted dish. Photography indicated an orthokinetic response to light intensity, but sharp changes in light intensity had no detectable effect on miracidial behavior. At the lower temperature the response to a directed light beam was almost completely inhibited and the orthokinetic response could no longer be demonstrated.


Distribution, dilution and elimination of polychlorinated biphenyl analogs in growing swine. Growing swine were given 15 mg/kg po of purified polychlorinated biphenyl analogs and a commercial mixture. Backfat biopsies for 16-18 weeks exhibited two-compartment elimination kinetics when based on concentration. Total body fat was estimated from potassium-40 abundance, and the calculated total fat load indicated that most of the decline in residue concentrations was due to dilution by growth and expansion of the fat compartment. Redistribution in the fat was apparent for various analogs at different times, and tissue distribution varied significantly among analogs.


The effects of probenecid upon the individual components of indomethacin elimination. The effects of probenecid upon plasma profiles for indomethacin and upon the individual components of total plasma clearance have been evaluated in dogs, monkeys and rats. These three species cover the wide spectrum of species differences with respect to the relative contribution of renal, biliary and metabolic clearances to elimination of indomethacin. In dogs, in which biliary clearance is the major component of elimination, probenecid raised incremental values for area under the plasma curve [AUC] greater than 2-fold and completely obliterated the normal portal: peripheral gradient of drug after i.v. dosage. Biliary clearance, measured directly, was inhibited 70%. In rats, in which metabolic clearance is predominant, probenecid had no significant effect upon plasma clearance. In rhesus and spider monkeys, in which renal, metabolic and biliary clearances are all significant, as in man, areas under the plasma curve for indomethacin were more than doubled by probenecid, concomitant with profound depressions of both renal and biliary clearances. If plasma levels are a valid index of pharmacological availabiltiy and amount of drug secreted in bile a correlate of intestinal irritation, probenecid increases therapeutic ratio with respect to intestinal toxicity ca. 5-fold in spider monkeys and 10-fold in rhesus monkeys.


[Concomitant Prinzmetal angina and coronary angina. Angiographic study in 13 patients (author's transl)]. Is coronary spasm the explanation for the paroxysmal myocardial ischaemia of Prinzmetal angina? Although considered adventurous two or three years ago, this hypothesis has been supported by the publication of approximately 15 cases in which the cause and effect relationship seemed to be proven by spasm seen on coronary arteriography at the same time as a typical attack. The authors report 13 cases of this type (the first of which was published in 1972). Spasm occurred in a normal coronary system in 3 cases, whilst in 2 cases there were only non-stenotic plaques of atheroma. In 8 cases, spasm occurred at the site of or distant from a significant atheromatous lesion. In 8 cases, spasme involved the right coronary whilst in 3 cases the A.I.V. was affected and in 2, two of the three main coronary trunks. In the light of their experience, the authors stress two points: --in many cases, nitroglycerine alone is not sufficient to relieve the spasm, and only the in situ injection of papaverine had a constant effect; --it is essential to opacify both coronary systems during a typical attack in order to obtain proof of spasm.


Synthesis and antimicrobial activity of azasteroid-type compounds and related systems. Effect of hydrophilic and lipophilic groups on activity. Pyrazole-, pyrazolone- and isoxazole-containing systems were prepared from 3,4-dihydro-6-(hexyloxy)-1(2H)-naphthalenone, 3,4-dihydro-6-(hexadecyloxy)-1(2H)-naphthalenone,3,4-dihydro-6(2-dimethylaminoethyloxy)-1-(2H)-naphthalenone, 3,4-dihydro-7-hexyloxy-1(2H)-phenanthrone, and 3,4-dihydro-7-(2-dimethylaminoethyloxy)-1(2H)-phenanthrone. A number of compounds derived from 7, 8-dihydro-5(6H)-quinolinone were also synthesized and characterized. Both hydrophilic and lipophilic groups were incorporated into certain systems as well as cidal groups. The compounds were screened for their in vitro inhibitory activity against Bacillus subtilis and Pseudomonas fluorescens. Structure-acitivity relationships among the molecular systems are discussed.


Structure-activity relationships among the O-acyl derivatives of leucomycin. Correlation of minimal inhibitory concentrations with binding to Escherichia coli ribosomes. The synthesis, antimicrobial activity, and binding to ribosomes of leucomycin and leucomycin derivatives are described. In general, the binding of the leucomycins and the leucomycin derivatives to ribosomes correlated with their antimicrobial activity. Some 2'-O-acyl derivatives apparently underwent gradual hydrolysis during antimicrobial assays, for their binding to ribosomes was poor compared to their relatively good antimicrobial activies. Correlation between antimicrobial activity and binding to ribosomes, their molecular site of action, provides some insight into the nature of the active molecular moieties.


Picornaviruses: rapid differentiation and identification by immune electronmicroscopy and immunodiffusion. Immune electronmicroscopy (IEM) was used to identify human picornaviruses rapidly and to differentiate enteroviruses from rhinoviruses. Human sera, diluted 10- to 50-fold beyond the neutralisation endpoints for homologous virus, readily agglutinated C-type antigens of seven human picornaviruses. Human sera did not react by IEM with a control animal picornavirus. By IEM after acid treatment, differentiation of a human enterovirus from a human rhinovirus was possible. There was an excellent correlation between the results of IEM and immunodiffusion (ID) tests for the presence of antibody to human picornavirus group antigens. By ID, only one of 21 human sera reacted with one of six animal picornaviruses. Immune electronmicroscopy appears to be a sensitive and simple techinque for the detection of picornavirus C-type antigens, and may be useful for identifying viruses belonging to groups comprising many serotypes and sharing a common group antigen.


A sequential bacteriological and serological investigation of Rhesus monkeys immunised against dental caries with Streptococcus mutans. In a serial investigation of the effects of immunisation with S. mutans in rhesus monkeys maintained on a "human" type of cariogenic diet, the numbers of S. mutans in cervical plaque, crevicular-fluid washings, fissures of teeth, and in saliva were lower in immunised animals than in sham-immunised controls. Immunisation also caused a delay in initial colonisation and a slowing of the rate of colonisation with S. mutans. These bacteriological changes were associated with a reduction in the smooth-surface-caries score. No relationship was found between the presence of S. sanguis and caries, but there was an inverse relationship between the proportions of S. mutans and S. sanguis isolated. Increased titres of complement-fixing, haemagglutinating and precipitating antibodies to S. mutans were found in the sera of immunised but not of control monkeys. A significant increase in salivary haemagglutinating antibodies was not detected. The results suggest that immunisation with S. mutans causes an increase in serum antibodies and a reduction in the number of S. mutans in the oral flora, and that these are associated with a reduction in dental caries.


Water fluxes in nerve fiber.

Water fluxes in nerve fiber. The hydrostatic (Lp) and osmotic (LPD) filtration coefficients and the efflux rates of tritiated water were measured in the giant axon of Loligo vulgaris. The Lp was 8 to 14 X 10(-8) cm/sec/cm H2O and the LPD was two orders of magnitude smaller (3 to 6 X 10(-10) cm/sec/cm H2O). In axons whose diameter was approximately 500 micron, the time (t1/2) required for a reduction in the axonal labeled water activity to one half its initial value was 38 to 48 sec. The rate limiting structure for solute flux was made ineffective by (1) storing the axon in isosmotoc KF at 0-2 degrees C for one month to one year or by (2) fixing the axon in 2-4% glutaraldehyde for 3 to 7 hr. The criteria of ineffectiveness of the rate limiting structure for solute flux were (1) a reduction of LPD to immeasurably low values, (2) the absence of electrical properties characteristic of plasmalemma, and (3) a marked increase in the rate of efflux of Na22. In such impaired axons the Lp and the t 1/2 of tritiated water efflux were unaffected. This independence of solute and solvent flux in conjunction with the finding that the hydraulic conductivity determined by bulk osmotic and hydrostatic pressure gradients is not equivalent (i.e., LPD/LP less than 1) indicate that the rate limiting structures for solute and solvent flux are in series. Solvent fluxes appear to be surface-limited, not bulk-limited. We have been unable to resolve whether the surface structure involved in limiting solvent flux is the sheath (Schwann layer and adhering connective tissue) and/or the cortical layer of the axoplasmic gel.


Effect of temperature on nuclear membranes and nucleo-cytoplasmic RNA-transport in Tetrahymena grown at different temperatures. The effect of temperature on the nuclear envelope structure and the transport of total RNA and ribosomal subunits from nucleus to cytoplasm was examined in Tetrahymena cells propagated at two different temperatures. Freeze-etch electron microscopy of cells grown at 23 and 18 degrees C detects the emergence of smooth areas on the fracture faces of the nuclear membranes upon lowering the temperature below approximately 15 and approximately 12 degrees C, respectively. Coincident with these freeze-etch changes, a discontinuous decrease is observed in the nucleocytoplasmic RNA-transport; this is probably not due to a cease in RNA-synthesis. Below the thermotropic discontinuity observed in the transport of total RNA in 18 degrees-cells the nucleocytoplasmic transport of the small and large ribosomal subunits is equally retarded. Recent temperature studies on the endoplasmic reticulum membranes of Tetrahymena suggest that the freeze-etch changes in the nuclear membranes are induced by a thermotropic clustering of the membrane lipids. We conclude that this lipid clustering induces the permanent protein constituents in the nuclear envelope pore complexes to change from a relatively "open" into a relatively "closed" state thus causing the observed decrease in RNA-transport.


Effect of ionophore A23187 upon membrane function and ion movement in human and toad erythrocytes. Addition of 0.1-0.3 micronM A23187, a divalent cation ionophore, to human erythrocytes suspended in a 1.0 mM 45Ca2+ -containing buffer results in a small (approximately two fold) increase in [Ca2+]i, a significant decrease in osmotic fragility, and a decrease in intracellular K+ (100 mmoles/liter of cells to 70 mmoles/liter cells) without significant alteration of intracellular [Na+]. This decrease in [K+]i is associated with a significant decrease in packed cell volume and correlates directly with the observed alteration is osmotic fragility. Increasing extracellular K+ to 125 mM prevents the A23187-induced changes in osmotic fragility, K+ content and cell volume, but does not prevent the ionophore-induced uptake of 45Ca2+. Addition of 0.1-0.3 micronM A23187 to toad erythrocytes leads to an increase in 45Ca2+ uptake comparable to that observed in human erythrocytes, but does not alter osmotic fragility, cell volume or K+ content. Higher concentrations of ionophore (3.0-10.0 micronM) cause a 30- to 50-fold increase in 45Ca2+ uptake and concomitant change in K+ content, cell volume and osmotic fragility. These changes in cell properties can be prevented by increasing extracellular [K+] to 90 mM. The difference in sensitivity of the two cell types to A23187 is attributed to the presence of additional intracellular calcium  pools within toad erythrocytes that prevent an increase in cytoplasmic Ca2+ until Ca2+ uptake is increased substantially at the higher concentrations of A23187.


Effects of depot injections of retinyl palmitate on 7,12-dimethylbenz[a]anthracene-induced preneoplastic changes in rat skin. The preneoplastic skin changes usually induced by topical application of 7,12-dimethylbenz[a]anthracene (DMBA) to adult rat skin did not appear when animals were treated locally with depot im injections of high doses of retinyl palmitate (RP) prior to exposure to the carcinogen. The epidermal histology after RP-DMBA treatment was similar to that seen in areas exposed to RP alone. Keratinization was inhibited but there was no cellular atypia, evidence of cell injury, or mucous metaplasia. Other features were hyperplasia with acanthosis and thickened stratum granulosum, parakeratosis, intercellular edema, and loss of hair overlying the injection site. Ultrastructurally, the epidermal cells contained conspicuously fewer tonofibrils and increased dense chromatin, when compared to control cells. Skin changes observed following treatment of littermates with DMBA alone included the appearance of giant tumor cells, dyskeratotic cells, nuclear hyperchromatism, increased nucleocytoplasmic ratio, and pleomorphic nuclei and nucleoli. oss of desmosomes, increased tonofibrils, and defects in the basement membrane with epithelial projections into the dermis were also seen. These preneoplastic changes did not regress when application with DMBA was discontinued after 6 weeks; exposure to the carcinogen for longer than 6 weeks resulted in an exacerbation of the abnormal state. RP had profound effects on rat epidermis that interfered with the effects of a potent skin carcinogen. The mechanisms underlying the phenomenon have not been defined. The use of depot injections of the vitamin which avoids both systemic toxicity and the local irritation seen with topical exposure could serve as a model in which the anticarcinogenesis properties of retinoids could be explored.


Autoradiographic and histopathologic studies on the mode of action of an aromatic retinoid (Ro 10-9359) on chemically induced epithelial tumors in Swiss mice. The mode of action of an aromatic analogue of retinoic acid, ethyl all-trans-9-(4-methoxy-2,3,6-trimethyl-phenyl)-3,7-dimethyl-2,4,6,8-nonatetraenoate (Ro 10-9359), a compound known to possess a considerable prophylactic and therapeutic effect on skin papillomas and carcinomas, was investigated with autoradiographic and histopathologic methods. The ip application of a single dose of 1,000 mg Ro 10-9359/kg to female Swiss mice with chemically induced skin papillomas caused a 29% regression of the mean tumor diameter after 3 days and a 51% regression after 7 days. In the tumors, the number of DNA-synthesizing cells [measured by the labeling index (LI)] and the length of the cell cycle were not affected by the retinoid; thus a mode of action at the level of cell proliferation can be excluded. In the normal skin, an increase in the LI of about 30% was observed. A small effect on the cell loss was observed; however, it was not sufficient to explain quantitatively the regression of the tumors. When measured histometrically, it appeared that the loss of the horn and the formation of necroses, 3-10 times larger than in the placebo groups, were mainly responsible for the tumor regressions caused by the retinoid. After 7 days, the proportion of stroma in the tumors was increased, and dilation of the vessels and edema in the stroma proximal to the necroses were frequent.


Early experience in using and 18 Me V linear accelerator for mycosis fungoides at Howard University Hospital. This paper describes the problems and solutions in using 18 MeV linear accelerator, with minimum 6 MeV electron capability, for total skin irradiation for mycosis fungoides. The 6 MeV electron energy can be degraded to acceptable electron energy of 3.2 MeV by interposing a plexiglass sheet of 9.6 mm in the beam. To minimize the bremsstrahlung, the degrading plexiglass should be kept away from the machine head. A wide area with uniform dose distribution over single plane can be achieved by using dual fields but homogenous dose distribution over irregular body surface cannot be achieved mainly because of self-shielding. The nails and the ocular lens can be easily shielded from the low energy electrons with 1.5 mm lead shield.


Stable and short-lived isotopes in the study of tissue distribution. Two relatively novel techniques that are utilized in the study of tissue distribution of drugs are discussed. The first technique, GC-MS fragmentography, provides an opportunity for the identification and quantification of several compounds and internal standards, in the same analysis, at picogram to nanogram amounts. The second technique utilizes short-lived radioisotopes and external scintigraphy. This approach can continuously monitor the tissue and organ distribution of an appropriately labeled compound. It produces quantifiable results and provides an opportunity for doing several experiments on the same animal in a noninvasive manner.