Inhibition of intravascular fibrin deposition by dipyridamole in experimental animals. Intravascular fibrin deposition was induced in rabbits by endotoxin, the infusion of fibrin monomer (FM), and by the infusion of thrombin and EACA. A previously developed radioisotope technique was used to measure the fibrin deposits in various organs. Dipyridamole treatment of rabbits caused significant inhibition of fibrin deposition in all three experimental models. The drug also inhibited platelet consumption and, in the thrombin- and EACA-infused animals, fibrinogen consumption as well. The results obtained with dipyridamole were compared with the effect of thorotrast. It was concluded from this comparison that the effect of dipyridamole could not be attributed to inhibition of the reticuloendothelial system. It is postulated that dipyridamole inhibits the final step at which soluble FM is precipitated as fibrin in vivo.


Separation of rosette-forming cells with the aid of an immunosorbent based on sephadex G-25 and G-75. A method of separating rosette-forming cells on columns with an immunosorbent based on sephadex G-75 is described and has been tested. The protein antigen (bovine serum albumin-BSA) is attatched by covalent bonds to the surface of the Sephadex granules oxidized by sodium periodate. Cells with receptors on their surface were tested by the rosette method. Up to 88% of the rosette-forming cells were retained on a column packed with BSA-Sephadex granules. Nonspecific retention of the cells was relatively small, about 4%.


A comparison of one layer and two layer techniques for colorectal anastomosis. In a randomized prospective clinical trial and layer and two layer techniques have been compared in 92 patients undergoing colorectal anastomosis. The results were assessed radiologically by barium studies on the tenth postoperative day. There was no significant difference in the incidence of anastomotic breakdown with either technique when the anastomosis was performed above the pelvic peritoneal reflection. When the anastomosis was situated below the pelvic peritoneum the incidence of dehiscence was significantly greater when a two layer technique was used.


Cervical spinal interbody fusion with Kiel bone. Experience with the use of the Kiel bone graft in anterior cervical interbody fusion in cervical spondylosis is reported. The survey reviews the results of operations on 73 disc spaces in 65 consecutive patients, from 2  to 5 years after the operation. We have failed to confirm that any bony fusion in the true sense occurs with Kiel bone in anterior cervical interbody fusion. However, the aims of the operations are achieved, namely removal of the disc and osteophyte, fixation of the spine to prevent compression of the neural tissues, distraction of the adjacent vertebral bodies providing increased room in the intervertebral foramina and stability of the spine.


IgE antibodies to Dermatophagoides pteronyssinus (Housedust mite), Aspergillus fumigatus, and beta-lactoglobulin in sudden infant death syndrome. The prevalence of serum IgE antibodies to Dermatophagoides pteronyssinus (house-dust mite), Aspergillus fumigatus, and bovine beta-lactoglobulin was significantly greater in cases of sudden infancy death (S.I.D.) than in a control group of infants of the same age range. This difference was most pronounced with D. pteronyssinus antibodies, which suggests that hypersensitivity to house-dust mite may be a factor in the aetiology of S.I.D. in Western Australia. Both the S.I.D. and control infants had similar serum concentrations of immunoglobulins G, M, and E but IgA levels were significantly higher in the control group.


Effects of ambulance transport in critically ill patients. Two groups of critically ill patients were transferred by ambulance from other hospitals to a central intensive therapy unit. The effect of transport was reviewed retrospectively in 46 patients and prospectively in 20 patients. Of the 46 patients reviewed retrospectively six became hypotensive, six became hypertensive, and seven developed delayed hypotension. One patient developed fits and six out of 13 patients had a rise in arterial PCO-2 of 1-6-4-1 kPa (12-31 mm Hg). Of the 20 patients reviewed prospectively, one patient became hypertensive due to overtransfusion, one had a fit, but none became hypotensive. Three out of four cases of delayed hypotension were related to starting intermittent positive pressure ventilation. Arterial PCO-2 fell in one patient and arterial PCO-2 rose in two, each change being related to changed oxygen therapy or narcotics. There were no changes in other cardiovascular or respiratory indices, body temperature, or urine production. Earlier transfer, resuscitation before transfer, continuing medical care during the journey, and hence a slower smoother journey seemed to be important factors in the management of these patients. Our findings, may have important implications in the future regional organization of the care of critically ill patients.


Immune complexes in cystic fibrosis.

Immune complexes in cystic fibrosis. Circulating immune complexes were detected in serum and sputum of patients with cystic fibrosis (C.F.). There were extensive deposits of immunoglobulins and complement immune complexes in several of the C.F. organs, especially the respiratory and gastrointestinal tracts, but not in the kidneys. Significant concentrations of IgG and of complement complexes could be eluted from the lungs of the C.F. patients but not from those of controls. Studies involving immunoabsorption, autoradiography, and molecular sieving through Sephadex G-200 columns identified both bovine serum albumin and staphylococcal alpha-haemolysin as two of the antigens present in the immune complexes. The sedimentation constant of the immune complexes was about 8S to 11S. The clinical significance of these immune complexes and the wide variety of antibodies detected in C.F. patients are discussed.


Comparison of aerosol ipratropium bromide and salbutamol in chronic bronchitis and asthma. The effects of inhaling 200 mu g of salbutamol were compared with those of inhaling 40 mu g of ipratropium bromide singly and in combination with salbutamol in eight patients with bronchitis and eight asthmatic patients in a double-blind controlled trial. Changes in airways resistance were assessed by measuring the forced expiratory volume in 1 second and specific airways conductance. Both drugs were significantly better in relieving airways obstruction than placebo. Salbutamol was significantly more effective than ipratropium bromide in patients with asthma, but in the patients with bronchitis there was no significant difference between salbutamol and ipratropium bromide. The combination of the two drugs produced a slightly greater and longer response than either drug alone but this was not significant.


Diseases causing end-stage renal failure in New South Wales. The nature of the original renal disease was determined in 403 consecutive cases of end-stage renal failure, in 317 of which the clinical diagnosis was corroborated by histological examination of the kidney. Five diseases accounted for 20 or more cases--glomerulonephritis (31% of the total), analgesic nephropathy (29%), primary vesicoureteral reflux (8%), essential hypertension (6%), and polycystic kidneys (5%). In only four cases did renal failure result from chronic pyelonephritis without a demonstrable primary cause. Greater use of micturating cystography and cystoscopy and routine urine testing for salicylate are advocated for earlier diagnosis of the major causes of "pyelonephritis". The incidence of end-stage renal failure in people aged 15-55 in New South Wales was estimated to be at least 34 new cases per million of total population each year.


Inhibition and amplification of the radiation-induced degradation of DNA in Escherichia coli. The degradation of DNA in a repair-proficient strain (B/r) and in two repair-defective strains (Bs-1 and pol A) of Escherichia coli was studied in the presence and absence of three metabolic inhibitors after exposure of the cells to X-radiation. The radiation-induced degradation of DNA was dependent on energy production in freshly harvested log-phase cells and was closely related to the repair capacity of the cells. A different system, probably involving nonspecific deoxyribonucleases acting at radiation-induced strand breaks, appeared to be responsible for the degradative processes in stationary-phase cells or in log-phase cells which had been aged by standing in buffered saline for 2 days before irradiation. The three inhibitors tested (caffeine, rifampin, and carbonyl cyanide m-chlorophenyl hydrazone) were all found to inhibit partially the radiation-induced degradation of DNA in E. coli cells. Under other conditions, the same three compounds amplified the degradative process. The data suggested that the amplification was due to a selective inhibition of repair processes. The degradative processes which were stimulated by X-radiation of E. coli cells show many parallels to those which are evoked by phleomycin or colicine.


The ecological significance of sinking to planktonic bacteria. Morphological and density measurements were made on planktonic bacteria from a subalpine lake. The observations were then used to predict in situ sinking rates from certain theoretical considerations. The predicted rates agreed well with observed sinking rates (on the order of 1 mm day-1). A simple analysis showed that sinking terms can be neglected in explaining distributions of unattached bacteria, except perhaps in relation to the presence of "bacterial plates."


Lysosomes and the "toxicity" of Rickettsias. VI. In vivo response of mouse peritoneal phagocytes to L-cell-grown Chlamydia psittaci 6BC strain. The L-cell-grown 6BC strain of C. psittaci inoculated intraperitoneally in mice induced an injurious effect on mononuclear phagocytes and their lysosomes; the influx of polymorphonuclear phagocytes (PMN's) increased markedly and the PMN's showed karyorrhexis and lysis. Cytochemical methods failed to detect chlamydial forms in peritoneal fluids from day 1 and up to 6 days after inoculation of mice. Chlamydial infectivity was not detected in either the cell-bound or the cell-free fractions of peritoneal cells from 6 h up to 6 days after inoculation of mice. In contrast, after infection of mice with the egg-grown 6BC strain, readily identifiable multiplication of the parasites occurred from 4 days after inoculation of mice; the lysosomes of macrophages were refractory to egg-grown 6CB particles that were initially ingested and the macrophages were nonspecifically "activated." Some of these "transformed" into large epitheloid cells containing numerous chlamydial inclusions in the cytoplasm. That lysosomes of "professional phagocytes" play a role in the "toxic" effect of virulent chlamydiae in the intact host is discussed.


A morphological study of anaerobic bacteria from the hypolimnia of two Michigan lakes. Dense populations of anaerobic bacteria were found sequentially layered below the thermocline in two eutrophic lakes in southwest Michigan. Phase and electron microscopy of whole cells and thin sections were used to reveal the in situ morphology of the dominant members of the community. The predominant chlorophyll-containing bacteria were identified on the basis of their morphology to be members of the genera Pelodictyon, Prosthecochloris, Clathrochloris, Chlorochromatium, Pelochromatium, Thiopedia, Thiocystis, Thiospirillum, and Chromatium. The natural morphology of these organisms is described and compared with the morphology of reported isolates, the morphology of unisolated genera was compared with previous descriptions of natural samples. Most of the organisms near the sediment-water interface and two from the upper hypolimnion have not been previously described. They have been divided into six distinct groups based on morphology; the morphological features of each group are presented. This approach, based on the morphological uniqueness of the procaryotes present, provides a satisfactory method for grouping members of the hypolimnetic community for ecological studies.


The structure of anaerobic bacterial communities in the hypolimnia of several Michigan lakes. The structure of bacterial communities, the distribution of sulfide and oxygen, bacteriochlorophyll concentrations, and the temperature profile were determined for the anaerobic hypolimnia of two lakes in southern Michigan. Information from these studies, plus  qualitative observations of two other lakes and two ponds over a 4-year period were used to correlate the spatial distribution of the populations, cell size, arrangement of photosynthetic vesicles or lamellae, presence of gas vacuoles or flagella, sulfur deposition, and environmental factors. On the basis of these results, three communities designated as A, B, and C were defined. The upper (A) community consisted of sequentially layered purple sulfur bacteria including two or more of the following genera: Thiopedia, Thiospirillum, Thiocystis, or Chromatium. The middle (B) community consisted of sequential layers of green bacteria from one or more of the following genera: Pelodictyon, Clathrochloris, Chlorochromatium, or Prosthecochloris. The lowest (C)community contained previously unreported gas-vacuolate colorless bacteria 0 to 0.7 m above the sediment. Microstratification (0.1- to 0.2-m layers) of populations was observed within the A and B communities.


Effect of chlorpromazine on the respiration and hexose monophosphate dehydrogenases of gram-negative bacteria. The drug chlorpromazine inhibits respiration in several gram-negative bacteria. The enzymes glucose-6-phosphogluconate dehydrogenase and 6-phosphogluconate dehydrogenase from these microorganisms are not affected by the drug.


Some aspects of wound healing research: a review. Mammalian response to injury essentially is that of tissue repair and re-epithelialization. The most important component of repair tissue is collagen, and after injury collagen turnover is greatly increased. Collagen biosynthesis is initiated by nuclear DNA of fibroblasts; the steps in biosynthesis are complex but studies of collagen biosynthesis may eventually have clinical potential. Normally, wound healing lasts for up to 2 years but nutritional and metabolic factors, such as malnutrition, delay healing; hyperalimentation would likely be beneficial under these conditions. Other factors that influence wound healing are the oxygen tension in tissues, the hemodynamic status, and the effects of substances such as cortisone, vitamins A and C, and zinc.


A double-blind comparison of intensive course 5-flourouracil by oral vs. intravenous route in the treatment of colorectal carcinoma. This radomized double-blind study was designed to compare the therapeutic effectiveness of the oral and i.v. routes for 5-FU administered in intensive courses to 100 patients with metastatic adenocarcinoma of the large bowel, treated to equivalent levels of toxicity. An oral dose of 20 mg/kg day times 5 was found to produce comparable G.I., mucocutaneous, and hematologic side effects to a dose of 13.5 mg/kg day times 5 by rapid i.v. injection. Courses were repeated at 5 weeks. Nine of 47, or 19.1%, treated by the oral route have shown objective response, compared to 14 of 53, or 26%, treated by the i.v. route. If malignant hepatomegaly is considered alone, the response rates are 8 of 23, or 34.8%, by the oral route, and 7 of 22, or 31.8%, by the i.v. route. The mean duration of response for the oral group, 11.1 weeks, was shorter than for the i.v. route, 20 weeks, a statistically significant (p less than 0.02) difference. Serial serum 5-FU levels after two doses of 5-FU were determined by microbiological assay in 19 patients. For i.v. administration the curves were comparable among different patients as well as in the same patient. There was striking variability, however, for oral administration.


Long-term remission in diffuse histiocytic lymphoma treated with combination sequential chemotherapy. Seventeen patients with Stage III or IV reticulum cell sarcoma were treated with three cycles of a 5-drug regimen between February, 1969 and December, 1970. Of the 17 patients, 9 attained a complete remission and 6 had a parital remission; 2 were considered unevaluable. Recently, the original biopsies from these patients were reclassified according to the criteria of rappaport et al. The results were: diffuse histiocytic--8; nodular histiocytic--2; diffuse mixed--2; nodular mixed--3; diffuse lymphocytic poorly differentiated--1; nodular lymphocytic poorly differentiated--1. Of the 8 patients with diffuse histiocytic lymphoma 6 attained complete remission, 1 had a partial remission, and 1 was unevaluable. One of the 6 with complete remission relapsed at 7 months and died 2 months later. However, the other 5 are alive and in continued unmaintained remission for 55 to 65 months.


Feminizing interstitial cell tumor of the testis: personal observations and a review of the literature. A 28-year-old man with evidence of feminization was demonstrated after 4 years of investigation to have a estrogen-secreting interstitial cell tumor. Such feminizing neoplasms are uncommon, only 37 having been described. They are usually benign and are characterized by gynecomastia, a testicular mass and, with lesser frequency, by decreased libido and potency and poor semen quality. The urinary excretion and plasma levels of estrogen are increased and, by selective testicular catheterization, the site of increased estrogen production can be localized. Secondary histologic changes occur in the nontumorous portions of the testis as well as in the contralateral testis; they are most marked in the area immediately adjacent to the tumor. Postoperatively, the gynecomastia regresses, the excessive levels of estrogen return to normal, libido improves, and the sperm count increases to normal.


Impact of the National Cancer Act on grant support. The National Cancer Act of 1971 resulted in a threefold increase in appropriations for the National Cancer Institute (NCI) within a 4-year period. A major effect was the increase for the Grants Program from +93 million in fiscal year 1970 to more than +280 million in 1974. Grant programs, administered by the Division of Cancer Research Resources and Centers, account for more than 50% of the total NCI extramural research budget and fall into four broad categories: research, training (including fellowships), cancer control, and construction. With the exception of the training area, funding for all grant programs has increased dramatically as a result of the Act. The ocst of research has also risen, as reflected in the average twofold increase in cost per NCI traditional grant over the past 10 years. This rise in cost is due to a number of factors, including inflation, more sophisticated equipment and supplies and, in some cases, more ambitious projects. The principal type of research grants include traditional awarded for investigator-initiated research projects, and center, awarded for comprehensive and specialized cancer centers. While support for traditional grants has remained in the forefront of NCI funding, money for cancer center grants has increased at a greater rate in recent years, reflecting emphasis on the development of cancer centers throughout the country. Compared to other institutes at the NIH, NCI is in a very favorable funding position; in fiscal year 1974 NCI awarded more money for its research grant programs than all of the other institutes (with the exception of the National Heart and Lung Institue) obligated for their entire budgets. The Act has stimulated a large increase in new cancer applications received, and the increased funding has made it possible for NCI to award a greater number of grants. Young investigators have competed well for the additional monies made available by the Act and funding for cancer research outside the United States, still only a small part of NCI's budget, has increased.


Quantitation of cell shedding into efferent blood of mammary adenocarcinoma. The rate of tumor cell shedding into efferent tumor blood was measured in growing and regressing MTW9 rat mammary carcinomas. The hormone-dependent tumor, grown as an 'isoession was induced by reduction of mammotropin level in the host. Tumor cells were differentiated from normal leukocytes by indirect immunofluorescence. Growing tumors shed 3.2 x 10-6 and regressing tumors shed 4.1 x 10-6 cells per 24 hr per g tissue. Cell shedding rates of growing versus regressing tumors were not siginificantly different over a tumor size range of  2 to 4 g. The number of tumor cells in the arterial blood was 12-fold smaller than in the efferent tumor blood. It is concluded that: (ay cell shedding via blod probably plays only a minor role in the total cell loss by gtowing MTW9 carinomas; (b) hormone-induced tumor regression does not depend on increased cell shedding; (c) tumor cells are rapidly cleared from circulating blood; and (d) a 2-g MTW9 carcinoma pours enough cells into the host circulation to transplant the tumor every 24 hr.


In vitro metabolism and microsome-mediated mutagenicity of dialkylnitrosamines in rat, hamster, and mouse tissues. Rates of conversion of 14C-labeled dimethyl-and diethylnitrosamine by rat and hamster tissue slices to 14CO2 and/or into mutagenic reactants were measured using Salmonella typhimurium G-46 r TA 1530 and fortified tissue fractions in vitro. A correlation between the CO2 production from dimethyl- or diethylnitrosamine in liver or lung and the organ distribution of induced tumors in vivo was observed. As an exception, hamster lung, which is a major target organ in diethylnitrosamine carcinogenesis, did not convert this nitrosamine into metabolites mutagenic for S. typhimurium TA 1530 although the 14CO2 production in vitro was even higher than in hamster liver. The effect of pretreating rats, hamsters, and mice with phenobarbitone on the mutation frequency produced by dimethyl-or diethylnitrosamine in in vitro assays was determined. The relationship between the site of metabolic activation, mutagenicity, and carcinogenicity of the dialkylnitrosamines and the effect of enzyme inducers are discussed.


Treatment of runting syndrome and prevention of primary lymphomas in friend virus-tolerant rats. A series of experiments showed that the inoculation of spleen and lymph node cells from rats immunized with Friend lymphoma (WFT-13) CELLS INTO Friend virus-tolerant rats induced the runting syndrome in nearly all cases, and immunological tolerance to WFT-13 was not broken in any survivors. However, th inoculation of specific immune spleen and lymph node cells admixed with normal bone marrow cells suppressed the runting death. In addition, in these animals the primary lymphomas that ordinarily occur about 200 days after neonatal inoculation of Friend virus did not appear. The mixture of immune spleen and lymph node cells and normal spleen and lymph node cells or normal thymus cells was ineffective in preventing the runting death or the incidence of primary lymphoma. Spleen and lymph node cells from normal rats or rats immunized with antigenically different AH-66 cells were also without effect. Spleen and lymph node cells from rats immunized with sheep red blood cells had a relatively high incidence of the runting syndrome; a few survivors rejected the WFT-13 transplants and also did not develop primary lymphomas. These results suggest that a supplement of hematopoietic stem cells from bone marrow eill not only prevent the runting death of Friend virus-tolerant rats produced by inoculating immune lymphoid cells but will also prevent the expected occurrence of primary lymphomas.


Epithelial and muscle antigens in benign cystic teratomas of ovary. Thirty beign cystic teratomas of the ovary were studied by immunofluroesecence, using specific human antisera for (a) squamous epithelial intercellular areas (pemphigus sera), (b) squamous epithelial basement membranes (bullous pemphigoid sera), (c) smooth muscle (chronic active hepatitis sera, and (d) striated muscle (thymona sera). Squamous epithelial intercellular staining was seen in all specimens and was present in the basal cell layers as well as in the prickle cell layers, suggesting a slow regeneration rate for the squamous epithelial lining of these tumors. Well-formed basement membranes were identified at the periphery of all squamous epithelial structures. Smooth muscle was found in 73% of the neoplasms, and the immunoflurorescent findings correlated with light microscopy. No skeletal muscle components were identified. Within the limits of the antibodies used and the antigens detected, these resutlts support the concept that benign ovarian cystic teratomas are composed of mature, heterotopic tissue elements.


Selection for high immunogenicity in drug-resistant sublines of murine lymphomas demonstrated by plaque assay. The immunogenicity of lymphoma L1210 and three L1210 sublines, resistant to methylglyoxal bis(guanylhydrazone), 4,4'-diacetyldiphenylurea bis(guanylhydrazone), or guanazole (L1210/GZL), respectively, was evaluated. Syngeneic DBA/2J mice were given a single i.p. injection of serially diluted suspension of irradiated cells from L1210 or L1210 sublines. Five days later spleen cells from the immunized mice were tested for the presence of plaque-forming cells using the immunizing lymphoma cell lines as target. Sera collected from the animals were examined for cytolytic antibody activity by lysis in gel using the same target cells. For comparison, the H-2 immunogenicity of L1210 and its sublines was investigated in H-2-incompatible allogeneic mice. The following results were obtained. (a) All the sublines showed increased immunogenicity and susceptibility to lysis as compared to L1210 cells. The number of plaque-forming cells/spleen ranged from 100 for L1210 to 4450 for L1210/GZL, the most immunogenic subline, and the antibody titer ranged from 1/8 for L1210 to 1/128 for L1210/GZL. (b) All the sublines carried common tumor-associated antigens that apparently made primary contributions to the increased immunogenicity. (c) The common tumor-associated antigens were also expressed on L1210 cells, although in a lesser defree, as evidenced by the definite, albeit low, capacity of L1210 cells to absorb DBA/2J anti-L1210/GZL antibodies. (d) Spleen and thymus cells of DBA/2J mice as well as unrelated murine ascites tumor cells did not cause significant absorption of these antibodies. (e) Only a partial inverse relationship could be demonstrated between tumor-associated antigens but the lowest for H-2. The above results would seem compatible with the hypothesis that the increased immunogenicity of drug-resistant L1210 sublines is attributable to the selection of preexisting highly immunogenic cells during immunosuppression by treatments selecting for drug resistance.


Comparison of the allospecific and viral-specific immune responses to irradiated versus formaldehyde-fixed allogeneic Moloney lymphoma cells in CBA mice. Two groups of adult CBA mice were immunized with 10-7 allogeneic Moloney lymphoma (YAC) cells. These YAC (H-2a) cells, which were either irradiated with 6000 R (Group i) or were formaldehyde fixed (Group II), were injected i.p. at weekly intervals for 3 weeks. Four days following the last injection, sera and lymphocytes were collected and tested in vitro for activity against either allospecific antigens (H-2d target cells) or viral-specific antigens, namely, Moloney leukemia virus (MLV). Both groups of animals developed measurable cellular and humoral immunity to the virally determined antigens. However, only the animals in Group i, immunized with irradiated cells, developed detectable immunity to H-2d. Immune and control lymphocytes were tested in microcytotoxicity tests and by 51Cr release. Antibody was assessed by complement-dependent cytotoxicity, indirect membrane immunofluorescence, virus neutralization, and antibody-dependent lymphocyte cytotoxicity. Group I serum, which had both anti-MLV and anti-H-2 antibodies, was absorbed with either living or formaldehyde-fixed YAC cells. The living cells were able to remove both H-2 and MLV antibodies. On the other hand, the formaldehyde-fixed cells removed no H-2 antibody but were able to remove MLV antibody, although less efficiently than living cells. These data indicate that formaldehyde fixation selectively impaired the H-2 antigens, leaving the viral antigenicity relatively intact. Differences between the immune responses to MLV-determined antigens and to H-2 antigens were demonstrated in many of the parallel in vitro tests.


Intermolecular linking and fragmentation of DNA by beta-propiolactone, a monoalkylating carcinogen. Brief exposure to beta-propiolactone (BPL) increases the sedimentation rate of purified Escherichia coli DNA in neutral and alkaline sucrose gradients. However, when electrophoresed in polyacrylamide-agarose gels, this BPL-treated DNA moves ahead of the control. Longer incubation with BPL gives rise to two new fractions, the first one sedimenting as a heterogeneous material of 6-8S, and the second one of very high sedimentation velocity. In acrylamide-agarose gels, the first fraction is again recovered in the 6-8S area, while the second fraction does not enter the gel at all. The DNA at this stage is hyperchromic in ultraviolet light suggesting that as much as 20% may be denaturated. Coliphage lambda DNA treated briefly with BPL and spread in a protein monolayer appears under the electron microscope as a rigid, extended molecule, up to 15% longer than the control DNA, and usually in compact, folded configurations suggesting intramolecular linking. After longer exposure, localized denaturation associated with single-strand breaks is observed. The single-stranded "whiskers" then interact with other DNA molecules, creating highly complex branched networks of single- and multi-stranded DNA. The possible relevance of these observations to the mechanisms involved in carcinogenesis and mutagenesis is considered.


Red cell carriage of label: its limiting effect on the exchange of materials in the liver. The red cell membrane is a permeability barrier that limits the equilibration of a variety of solutes between red cell and plasma water. We utilized the multiple indicator dilution technique to investigate the effect of this barrier on the exchange in the liver of a group of tracer substances that are not removed in net fashion from the hepatic circulation: thiourea, urea, and chloride. We demonstrated that, after preequilibration of the label with red cells, a red cell carriage effect appeared (the trapping and translocation of label in the red cells), that this effect was most marked when the permeability of the red cell was relatively low for the substance under consideration (thiourea), and that the effect became small when the permeability of the red cells was large for the exchanging substance (urea and chloride). We developed a theoretical description of the retarding effect of the red cell permeability barrier on the extravascular exchange of label and were able to use this description to obtain estimates of the red cell permeability from the in vivo dilution curves. We examined the effect of plasma injection, of changing the input in such a fashion that the label was not preequilibrated with red cells, and found both experimentally and theoretically, that for substances of low permeability the transit time from these experiments, if multiplied by the total water flow or solute flux, gave an overestimate of both the apparent total volume of distribution and the mass of traced material in the system. This last effect is of great importance for the practical design of many biological experiments. Reliable volume and mass estimates can be made only when the labeled material has been preequilibrated with red cells.


Red cell permeability effect on the mean transit of an indicator transported through an organ by red cells and plasma. When an indicator that permeates red blood cells with an equilibration time of the order of the mean transit time of the indicator through an organ is bolus injected solely into the plasma inflow to that organ, the mean transit time of the indicator is greater than it is when the indicator is injected preequilibrated between the plasma inflow and the red cell inflow. The mean transit time for equilibrated entry is in turn greater than that  for indicator injected solely into the red cell inflow. A simplified method of calculation of this effect is given in the present paper; it requires only a model solution for the indicator steady state as distinguished from a more complicated time-dependent solution. The dependence of the effect on red cell membrane permeability allows this permeability to suitable indicators to be deduced. The method was applied to existing data on urea and thiourea passage through the dog kidney.


Echocardiographic features of total anomalous pulmonary venous connection. Seven patients, four months to ten years of age, with total anomalous pulmonary venous connection (TAPVC) were studied by echocardiography.  The diagnosis was confirmed by cardiac catheterization, angiography, and surgery in each.  The connection was to the left vertical vein in four patients, to the coronary sinus in one, to the inferior vena cava in one, and directly to the right atrium in one.  All patients exhibited the echocardiographic criteria of right ventricular diastolic volume overload (RVDVO).  In addition, an echo-free space was identified dorsal to the posterior wall of the left atrium.  Indocyanine green dye studies done in one of the patients provided evidence that this echo-free space represents the common pulmonary venous chamber.  The combination of echocardiographic evidence of RVDVO and demonstration of an echo-free space dorsal to the left atrium is strongly suggestive of TAPVC and should allow an early diagnosis in infants with this anomaly.


Circulatory and metabolic effects of glycerol infusion in patients with recent cerebral infarction. The effect of intravenous infusion of 10 per cent glycerol on regional cerebral blood flow (using hydrogen bolus and Xenon-133 (133Xe) clearance methods) and metabolism was investigated in 57 patients with recent cerebral infarction. Hemispheric blood flow (HBF) increased, together with increase in regional cerebral blood flow (rCBF) and cerebral blood volume (rCBV), in foci of brain ischemia. Hemispheric oxygen consumption (HMIO2) decreased together with hemispheric respiratory quotient. Systemic blood levels of glucose, lactate, pyruvate, and triglycerides also increased after glycerol while free fatty acids (FFA) and inorganic phosphate (Pi) decreased. Hemispheric glucose consumption was unaltered after glycerol so that hemispheric glucose to oxygen ratio tended to rise. Pyruvate and lactate production by brain was unchanged. Glycerol moved across the blood brain barrier into brain and cerebrospinal fluid (CSF). Release of FFA and Pi from infarcted brain was reversed by glycerol. Total phosphate balance was maintained actoss brain both before and after glycerol infusion. Triglycerides increased in CSF after glycerol, originating either from cerebral blood or as a result of lipogenesis in cerebral tissue. The EEG Recording and neurological status of the patients improved despite decreased brain oxygen consumption. Results of this study suggest that after intravenous infusion of 10 per cent glycerol in patients with recent cerebral infarction, glycerol rapidly enters the CSF and brain compartments and favorably affects the stroke process in two ways: first, by redistribution of cerebral blood flow with increase in rCBF and rCBV in ischemic brain secondary to reduction in focal cerebral edema; and second glycerol may become an alternative source of energy either by being directly metabolized by the brain, or indirectly, by enhancing lipogenesis, or by both processes. Involvement of glycerol in lipogenesis with esterification to accumulated FFA might lead to improved coupling of oxidative phosphorylation, a hypothesis that fits the finding of improved neuronal function despite further decrease in cerebral hemispheric oxygen consumption.


Serum zinc, iron, and copper concentrations during typhoid fever in man: effect of chloramphenicol therapy. In volunteers experimentally infected with Salmonella typhi, serum iron and zinc concentrations became significantly depressed and there was a concomitant rise in serum copper before the onset of overt clinical illness. However, after several days of fever and the initiation of chloramphenicol therapy, serum iron and zinc concentrations significantly increased. Additional studies--in volunteers with typhoid fever treated with chloramphenicol, in a volunteer with typhoid fever receiving cefazolin and gentamicin, and in untreated rhesus monkeys infected with Salmonella typhimurium--provided evidence that the increase in serum iron concentration during the febrile phase was the result of chloramphenicol therapy, whereas the increase in serum zinc concentrations was a disease-related phenomenon. The importance of trace-metal monitoring during infectious disease and chemotherapy is discussed.


Deformities of the midface resulting from malunited orbital and naso-orbital fractures. In midfacial fractures the crucial area is the upper portion of the midfacial skeleton which lodges the orbits, the interorbital space with its anatomic relationship with the anterior cranial fossa, the lacrimal apparatus, the levator muscle, and the skeletal structures of the nose. Because of the severity of the injuries in a multisystem injured patient, these fractures may be neglected or receive inadequate primary treatment. In some cases the damage is so extensive that primary treatment can only accomplish part of the task. Late reconstruction is necessary, therefore, and is the subject of this paper.


Alcohol involvement in highway crashes. A review of the epidemiologic evidence. There is probably no other single aspect of American behavior which is so underinvestigated as driving, and yet which has such an impact on our daily lives and on our annual death rate. Evidence is reviewed which indicates that the probability of being responsible for a fatal crash increases sharply as blood alcohol concentrations increase above those achieved by heavy social drinking.


Facial scars resulting from vehicular accidents. The surgeon will be better able to cope with the problem of scars as he understands the nature of wound healing and as advances are made in the knowledge of the physiology and biochemistry of tissues. Still there do remain the all-important careful handling of tissues, adequate wound repair, and adequate postoperative care of the wound and the patient.


Midfacial fractures from vehicular accidents. An effort is made to simplify, categorize, and outline the various forms and treatment of midfacial trauma from automotive crashes. Panfacial fractures resulting from high speed automotive accidents will continue to be a challenge to the innovative and persistent surgeon seeking the best possible results.


Soft tissue facial injuries related to vehicular accidents. Most facial injuries are not dire emergencies, except in the case of airway obstruction and in unrecognized, prolonged bleeding or oozing from facial wounds. Yet one must keep in mind the psychologic impact that facial injuries may have upon the patient and his family.


Frontal sinus and supraorbital fractures from vehicle accidents. Fractures of the upper third of the face from vehicle accidents are relatively uncommon but can have serious associated injuries of the dura and frontal lobe when they do occur. The injury may result from the victim's face either striking the dashboard or bouncing violently off the lower broken edge of the penetrated windshield.


A double-blind study in hypertensive patients of an original new compound, indapamide. In a double-bind crossover study in 18 patients with essential hypertension, the hypotensive activity of 5 mg. indapamide daily was compared with 40 mg. frusemide daily over a period of 4 months after an initial 15 days on placebo. The overall clinical assessment showed satisfactory blood pressure control in 72% of patients receiving indapamide compared with 57% on frusemide. The weight of patients on active therapy dropped significantly with both products, but to a greater extent with indapamide. Indapamide was well-tolerated by all 18 patients; 3 patients on frusemide developed side-effects. The results of blood chemistry investigations are discussed. Variations in potassium levels during indapamide therapy were modest and did not warrant the use of potassium supplements.


The treatment of acute anxiety states in neurotic patients with intravenous lorazepam. A placebo-controlled study. In a double-blind trial, 20 neurotic patients with acute anxiety were treated with 3 mg. lorazepan intravenously and 17 others were treated with placebo. A total of 24 injections of lorazepam and 26 of plter compared with 9 patients on placebo (p smaller than 0.05). Sixty minutes after the injections of lorazepam, patients were free of anxiety symptoms on 70.8% occasions, compared with only 7.7% occasions following placebo (p smaller than 0.01). The effects of lorazepam on anxiety were discernible 10 minutes after the injections. Considering the number of patients, not injections, 85.0% were free of symptoms after treatment with lorazepam and only 11.8% after treatment with placebo (p smaller than 0.01). Blood pressure, pulse rate and respiratory rate were favourably influenced by the drug and not by placebo; these effects were discernible 10 minutes after the injection.


double-blind"double-bind.

double-blind"double-bind. In a double-blind trial over a 2-week period in 34 patients with psoriases vulgaris, clobetasol propionate cream was shown to be more effective than betamethasone valerate cream. In a further 16 patients whose psoriasis had not responded to betamethasone valerate ointment, treatment with clobetasol propionate ointment was substituted. Fourteen of the 16 patients experienced a rapid improvement of their lesions.


Antihypertensive activity of a new agent, indapamide: a double-blind study. In a double-blind trail in 22 patients with mild to moderate essential hypertension, indapamide was compared with chlorathiazide and placebo. Dosage levels were set at 5 mg. indapamide and 500 mg. chlorothiazide daily, for 5 days out of 7, and patients were treated alternately, in random sequence, with each drug for a month over a 3-month period. Blood pressure readings and blood chemistry investigations were carried out before and after each treatment period and a careful history was kept of subjective symptoms and patients' tolerance of therapy. The results show that both active treatments produced drops in systolic and diastolic pressures, but were only statisically significant and different from placebo in the case of indapamide. Indapamide also produce much greater subjective improvement (74%) in patients with functional symptoms compared with chlorothiazide (15%). In an overall assessment, indapamide produced an excellent to good response to treatment in 57% of patients. Comparable responses for chlorotiazide and placebo were 25% and 20% respectivlely. Using patients as their own controls to compare the relative effectiveness of the three periods of treatment, indapamide was shown to be more effective than placebo in 65% of cases and more effective han chlorothiazide in 60%. Although patient tolerance of indapamide was slightly better, both drugs were well accepted and no significant changes from baseline levels were noted in any of the laboratory parameters investigated.


Dissociation between indices of pump performance and contractility in patients with coronary artery disease and acute myocardial infarction. An analysis of ventricular performance comparing pump function and muscle function indices was performed in 13 patients with acute myocardial infarction, ten patients with normal coronary arteries, and 15 patients with coronary artery disease. Pump function was described by plotting left ventricular stroke work index as a function of left ventricular end diastolic pressure. This description provided a clear separation between normal patients, and surviving and nonsurviving patients with acute myocardial infarction. Values of contractile element velocity (VCE5 as an estimate of Vmax) did not separate between normals and surviving or nonsurviving patients with acute myocardial infarction. In 15 patients with acute coronary artery disease there was no correlation between values of VCE5 and the ventricular function curve. Changes in performance following the stress of ventriculography, angiotensin infusion, or isometric hand grip exercise also did not show any correlation between pump function and muscle function indices. It is concluded that pump function indices are a better indicator of ventricular performance in patients with acute myocardial infarction and coronary artery disease.


High level positive end expiratory pressure (PEEP) in acute respiratory insufficiency. Twenty-eight patients developed severe, progressive acute respiratory insufficiency despite aggressive application of conventional respiratory  therapy. Application of increased PEEP (18 torr or greater) resulted in a significant decrease in QA/QT. Selection of the optimal levle of PEEP for each patient required serial determinations of QA/QT and measurement of cardiovascular response. The overall survival rate was 61 percent. Acute respiratory insufficiency was a proximate cause of death in only one patient. Four of the patients (14 percent) developed a pneumothorax following institution of high PEEP therapy. Cardiac output was not affected adversely at any level of PEEP up to 32 torr (44 cm H2O). We conclude that high levels of PEEP can be therapeutic for patients with refractory respiratory failure when combined with intermittent mandatory ventilation and careful cardiovascular monitoring. As with any therapy, the optimum dose should be tailored to each patient according to his needs and response.


Chicken soup rebound and relapse of pneumonia: report of a case. A case is reported in which a previously healthy individual, having received an inadequate course of chicken soup in treatment of mild pneumococcal pneumonia, experienced a severe relapse, refractory to all medical treatment and eventually requiring thoracotomy. The pharmacology of chicken soup is reviewed and the dangers of abrupt termination of therapy are stressed.


Bronchiectasis and homozygous alpha1-antitrypsin deficiency. A 34-year-old woman with homozygous a1-antitrypsin deficiency suffered from progressive, generalized cystic bronchiectasis. Although bronchiectasis was reported in the original monograph on the enzyme inhibitor deficiency, it has received minimal attention since then. Alpha1-antitrypsin levels should be measured in patients with severe bronchiectasis.


Extensive linear "blow-out" of the thoracic membranes trachea with innominate artery avulsion secondary to blunt chest trauma. The successful management of extensive rupture of the entire intrathoracic membranous trachea and an avulsed innominate artery is presented. Taken separately, the two entities are in themselves rare following blunt trauma to the chest. Ventilatory and anesthetic control during repair of this injury is presented.


Vector analysis of postcardiotomy behavioral phenomena. The classification of postcardiotomy behavioral phenomena in Figure 1 is proposed for use as a clinical instrument to analyze etiological determinants. The utilization of a vector analysis analogy inherently denies absolutism. Classifications A-P are presented as prototypes of certain ratio imbalances of the metabolic, hemodynamic, environmental, and psychic vectors. Such a system allows for change from one type to another according to the individuality of the patient and the highly specific changes in his clinical presentation. A vector analysis also allows for infinite intermediary ratio imbalances between classification types as a function of time. Thus, postcardiotomy behavioral phenomena could be viewed as the vector summation of hemodynamic, metabolic, environmental, and psychic processes at a given point in time. Elaboration of unknown determinants in this complex syndrome appears to be task for the future.


Pipotiazine palmitate: an evaluation of a new long acting intramuscular antipsychotic agent in severely ill schizophrenic patients. Findings in this study support earlier investigations in attesting to the antipsychotic efficacy and relatively low toxicity of pipotiazine palmitate. Results with all efficacy measures utilized were consistent in indicating a high level of efficacy for this investigational compound. Pipotiazine palmitate apparently has an average duration of action that extends beyond 4 weeks in severely ill schizophrenic patients. This particular long acting IM antipsychotic preparation appears to have an even longer duration of activity than some of the other available standard long acting agents. The optimal dosage range for severely ill schizophrenic patients appears to be between 100 and 600 mg once monthly. While this type of drug (as is the case with many antipsychotic drugs) does reduce the psychotic symptomatology and improves the thought associations sufficient to enable the patient to leave the hospital, it should be re-emphasized that socioeconomic and guidance counseling services are necessary to maintain the patient in the community. The availability of this type of long acting preparation is not only economical in terms of nursing care and hospital cost but it should also increase the efficacy of psychopharmacologic treatment of schizophrenics by reducing both patient errors and staff errors in administration of medication. In addition, this IM preparation should prove to be of invaluable help in maintaining the schizophrenic patient in his community by reducing the relapse and the rehospitalization rates. It should be noted that there are schizophrenic patients who either absorb compounds from the gastrointestinal tract in a very poor manner or too rapidly metabolize the antipsychotic agents with resultant suboptimal blood levels and these subjects may be called "drug refractory." This type of long acting medication is an ideal preparation for the schizophrenic patient who has these types of absorption or metabolic problems since the "circulatory pass" through the liver is minimal after IM medication as compared to that encountered by an orally administered agent. The clinical disadvantage of pipotiazine palmitate is the delay in onset of therapeutic activity after injection. Significant improvement is first noted after 3 to 4 days after the highest IM dosage administration. Therefore, it may be necessary to use an oral or IM preparation of a neuroleptic with a more rapid onset of activity or utilize an oral dosage of the pipotiazine salt during the first week following IM administration of pipotiazine palmitate.


[The effect of synthetic somatostatin in normal and acromegalic males]. The synthetic linear tetradekapeptide somatostatin (growth-hormone release inhibitory hormone: GHRIH) inhibits the liberation of growth hormone in normal persons in the insulin hypoglycaemia test without influencing the rise of cortisol and prolactin, while the concentrations of LH, FSH and TSH remain unchanged. In patients with florid acromegaly there occurs during administration of GHRIH a marked fall in the raised STH level without influencing the basal level of the other anterior-pituitary hormones. As a further effect there is suppression of the insulin level. The somatostatin at present available has a very short biological half-life and in its present form is, therefore, without therapeutic importance.


[Antinuclear factors in patients with pancreatitis of unknown aetiology: A pointer to an immunopathological cause? (Author's transl)]. Antinuclear factors in serum were determined (in addition to other tissue antibodies) in 9 patients with acute and 22 with chronic pancreatitis. Of the 11 with pancreatitis of unknown aetiology 9 had serum antinuclear factors, most of them in high titres. Of the 20 with pancreatitis of known aetiology only 3 had antinuclear factors.


[Amenorrhoea after oral contraceptives]. Eleven women with amenorrhoea after taking oral contraceptives were studied.  A clomiphene stimulation test was performed in ten, with six also having an LH-RH stimulation test.  In most of them there was no reaction or a severely impaired response to both clomiphene and LH-RH, but bleeding episodes could be provoked in most women by continuing clomiphene treatment, and some of these ovulated when HCG was added to the therapeutic scheme.  A possible relationship between preexisting psychosomatic disorders and the genesis of postcontraceptive amenorrhoea is discussed.


Effects of gonadal steroids on the basal and LRF-induced gonadotropin secretion by cultures of rat pituitary. The effects of 17beta-estradiol (E2), progesterone (P4), 20alpha-hydroxypregn-4-en-3-one (20alpha-OHP4), and testosterone (T) on the basal and LRF-induced secretion of LH and FSH were studies in monolayer cultures prepared from the pituitaries of adult female rats. Day 4 cultures were used and all steroids were tested at 10-8M concentration for 4 hr. E2 (2.72 ng/ml) alone caused a nonsignificant increase in basal secretion of both LH and FSH; however, the same dose of E2 significantly (p less than 0.001) inhibited the LRF-induced secretion of LH but not of FSH (74% and 88% of 10-8M LRF-treated level, respectively), Testosterone (2.88 ng/ml) alone significantly increased the basal secretion of LH (136% of control level, p less than 0.05) and augmented the effect of LRF on FSH secretion to 130% of the LRF-treated level (p less than 0.05). Contrary to its negative feedback action on the basal secretion of FSH (46% of control level, P less than 0.05), 20ALPHA-OPH4 (3.15 ng/ml) augmented the effect of LRF on LH secretion (130% of LRF-treated level, P less than 0.05). On the other hand, P4 (3.14 ng/ml) did not cause any significant inhibition in the basal and LRF-induced secretion of either LH or FSH. These data indicate that both of the LH- and FSH-LRF; however, their secretory activities are modulated differently by various steroids.


Radioimmunoassay for luteinizing hormone-releasing hormone (LHRH): its application to the measurement of LHRH in ovine and human plasma. A radioimmunoassay for LHRH, MODIFIED FROM THe assay of Nett et al. (J Clin Endocrinol Metab 36: 880, 1973) and characterized in more detail, was applied to the measurement of endogenous plasma LHRH in the peripheral circulation of ewes and women around the time of gonadotropin release. Sensitivity was 1.2 pg/tube. Within- and between-assay coefficients of variation were 10% and 25%, respectively, over the range 10-100 pg/tube. Minimal cross reaction (less than 0.1%) was observed with LHRH analogues tested, except those which had undergone single amino acid alterations in the 4 or 8 positions (62-93%) and in the 1 position (1-3%). Inhibition curves parallel to synthetic LHRH were obtained with these immunoreactive analogs, and with crude hypothalamic and pituitary extracts from 16 cycling ewes and 3 pooled hypothalamic extracts from male rats. LHRH contents of ovine hypothalami and pituitaries ranged from 1.9-10.2 (4.5 plus or minus 2.1, mean plus or minus SD) and 0-27.5 (4.2 plus or minus 7.3, mean plus or minus SD) ng LHRH, respectively, with no obvious correlation between the contents of either tissue. LHRH contents of pooled hypothalami from normal, castrated-hypophysectomized male rats were 4.5, 2.9 and 1. 3 ng/hypothalamus. Recovery of synthetic LHRH added to plasma was quantitative, provided the storage time at 4 and 20 C was minimal. Synthetic LHRH  administered to ewes by intravenous infusion or subcutaneous injection was readily detectable in peripheral plasma. The metabolic clearance rates of synthetic LHRH' in 3 sheep were 14, 11 and 14-1/day/kg, respectively. Endogenous immunoreactive material in single, unextracted plasma samples from sheep in various physiological states ranged from 0-400 pg/ml. However, the endogenous plasma immunoreactivity in these samples bone no resemblance to synthetic LHRH added to ovine plasma in that it was stable to gentle heat, and was undetectable after methanol extraction. No endogenous LHRH could be detected in methanol extracts of peripheral plasma obtained from estrous ewes or oophorectomized ewes injected iv with estradiol-17beta (40mug) and bled at 15 min intervals prior to gonadotropin release. Single daily plasma samples obtained from three normal women over their entire menstrual cycles contained no significant levels of endogenous LHRH.


Effects of estradiol-17beta on LH-RH/FSH-RH-induced, and spontaneous, LH release in prepubertal female pigs. The effect of estrogen (E2) on pituitary response to LH-RH/FSH-RH (LH-RH) was examined in immature female pigs (gilts). Eight gilts (9-10 weeks old) received iv injections of 2.5, 5.0, 10.0, 20.0, and 40.0 mug LH-RH, beginning 4 days after placement of indwelling jugular cannulae. Blood samples were drawn at 5-min intervals following LH-RH injection and plasma LH was determined by RIA and expressed as ng R786-3/ml. Pituitary responses were expressed as the increment in LH levels above baseline (delta LH). Pretreatment LH levels were 0.45-1.25 ng/ml. Responses to LH-RH (delta LH plus or minus SE) ranged from 3.7 plus or minus 0.6 (at 5 min) to 8.4 plus or minus 1.4 ng/ml (at 20 min) for 2.5 and 40 mug doses, respectively. Three additional prepubertal gilts were cannulated and implanted with E2. After 3 days plasma estrogens rose from approximately 30 pg/ml to 289 plus or minus 34 pg/ml and the dose-response studies were repeated. Prestimulation LH concentration was consistently below 0.4 ng/ml and delta LH levels ranged from 1.4 plus or minus 0.5 (at 10 min) to 5.7 plus or minus 1.3 ng/ml (at 20 min) for the 2.5 and 40.0 mug doses, respectively. Analysis of the regression equations for delta LH on log LH-RH dose showed that the adjusted means for LH response were significantly lower (P less than 0.02) in E2 implanted animals. However, the regression coefficients were not different (P greater than 0.5). On the basis of these and other analyses, it is concluded that chronic E2 treatment depresses both basal LH secretion and pituitary responsiveness, but that pituitary sensitivity to LH-RH is not modified. Several nonimplanted females showed bursts of LH release subsequent to the LH-RH induced peak, but multiple peaks were never observed in E2 implanted gilts. An explanation was sought and 7 of the nonimplanted gilts were used to study spontaneous LH release by determining plasma LH at 10-min intervals for 20 h. During this period spontaneous episodes of LH release were observed in control animals with a mean frequency of 1.3 LH peaks/h. When 4 animals were given 5 mug E2 iv and then sampled for 12 h, episodic secretion was suppressed and the latency to the first spontaneous LH release was 9.6 plus or minus 0.8 h, significantly greater (P less than 0.02) than the latency to the first peak after saline injections (4.2 plus or minus 1.5 h) in the controls. It is suggested that "multiple" LH peaks occurring after initial LH-RH stimulation may represent the superimposition of spontaneous episodic release upon the induced response.


Modulation of pituitary responsiveness to luteinizing hormone-releasing factor during the estrous cycle of the rat. This study was performed to investigate pituitary responsiveness to synthetic gonadotropic hormone-releasing hormone (Gn-RH) administered during the 5-day estrous cycle of the rat and to examine whether the observed differences can be attributed to ovarian secretion of estradiol. Gn-RH was injected intra-arterially at 4:00 PM to rats previously anesthetized with sodium pentothal to block the LH surge on proestrus and thus minimize changes in LH secretion which occur throughout the estrous cycle. Pituitary responsiveness was defined as the difference between serum LH concentrations in samples obtained immediately before and 15 min after administration of 200 ng Gn-RH, a time when maximal circulating levels of LH were observed. Administration of Gn-RH was followed by a significant increase in circulating LH on all days of the estrous cycle on which the response was tested (diestrus 2, diestrus 3, proestrus, estrus). Pituitary responsiveness was relatively low on diestrus 2 and estrus and was increased slightly on diestrus 3. On proestrus, however, pituitary response to Gn-RH increased markedly, a phenomenon abolished by ovariectomy at 8:00 AM on diestrus 3. The large increase in pituitary responsiveness observed on proestrus was not restored in such ovariectomized rats when circulating estradiol concentrations were increased and maintained at approximately 150 pg/ml by SC insertion of Silastic capsules containing estradiol-17beta immediately following ovariectomy. Nevertheless, this estradiol treatment consistently elicited an LH surge in another group of ovariectomized rats not treated with sodium pentothal or Gn-RH. Although these observations indicate that an ovarian hormone is essential for the increase in pituitary response to Gn-RH on proestrus, the identity of this hormone remains to be established.


Cortisol metabolism in the baboon during pregnancy and the postpartum period. The metabolism of iv-administered 14C-cortisol (F) by pregnant baboons (107, 124 and 150 days gestation) was compared with that previously reported for nonpregnant animals and with that of animals examined 6-18 h after spontaneous vaginal delivery (178 plus or minus 4 days). Unconjugated, glucuronoside (beta-glucuronidase) and sulfate (H2SO4-ethyl acetate) fractions were extracted with ethyl acetate from urine containing more than 80% of injected 14C. Metabolites of interest were isolated by paper partition chromatography and purified by crystallization and derivative formation. Compared with nonpregnant animals, the following changes (P less than 0.05) were observed in pregnancy: (1) an increase in the percent urinary 14C in the unconjugated fraction and a decrease in the proportion of 14C appearing in the glucuronoside fraction; (2) an increase in excretion of metabolites more polar than the cortols; (3) a decrease in excretion of metabolites less polar than cortisone in the glucuronoside fraction; (4) an increase in unconjugated F excretion. Production rate of F (11.9 plus or minus 0.7 mg/day) estimated by isotope dilution and percent urinary 14C in tetrahydrocortisol and tetrahydrocortisone from the glucuronoside fraction were as in nonpregnant animals. With the exception of an increase in F production (22.7 plus or minus 0.8 mg/day), presumably the result of the stress of labor, F metabolism in the immediate postpartum period was strikingly similar to timals indicates that changes in the mother alone can account for the altered metabolic disposition of F in pregnancy and suggests that the fetus takes little part in metabolism of maternal circulating F.


Ovarian modulation of immunoreactive gonadotropins-releasing hormone (Gn-RH) in the rat brain: evidence for a differential effect on the anterior and mid-hypothalamus. Gonadotropins-releasing hormone (Gn-RH) in selected regions of the female rat brain was measured by radioimmunoassay. Detectable immunoreactive Gn-RH was found in the anterior hypothalamic-septal region and in the mid-hypothalamic (arcuate-median eminence) region. Gn-RH was several times higher in the middle region than in the anterior region. Gn-RH was undetectable in the posterior hypothalamic region, frontal cerebral cortex and pineal glands, as well as in random blood samples, and low to undetectable in anterior pituitary glands. Gn-RH activity varied during the estrous cycle and after castration. In the mid-hypothalamic region, Gn-RH content was lowest throughout diestrus and in late morning and early afternoon of proestrus, and highest early in the morning of proestrus and during estrus. A significant decrease at mid-day was only found on the day of proestrus, a few hours prior to the critical period for LH release. In the anterior hypothalamic region, low Gn-RH activity was found from 1200 h of estrus to 1200 h of diestrus-2. A comparatively higher activity was seen at 1700 h of diestrus-2 and also from 1400 h of proestrus to 0800 h of estrus. Twenty-one days after ovariectomy, Gn-RH in the mid-hypothalamic region was significantly lower than the lowest values seen during the estrous cycle, while Gn-RH in the anterior hypothalamic region remained between low and high values seen during the cycle, being significantly higher than the low values. The changes observed during the estrous cycle and after castration suggest that gonadal steroids play a direct role in the control of hypothalamic Gn-RH. These data also demonstrate that Gn-RH varies in a different way in the anterior and mid-hypothalamic regions.


Dual action of antidepressant drugs (MAO inhibitors) on insulin release. Experiments have been carried out to investigate the role of monoamine oxidase (MAO) in the mechanism of insulin release. Isolated islets and pieces of rat pancreas were incubated in media of high glucose content in the presence of various concentrations of MAO inhibitors. At the end of the incubation, the islet MAO activity and the concentration of insulin released into the medium was measured. The results have shown that: 1. Glucose-mediated insulin release was potentiated by low concentrations of MAO inhibitors, the highest effect being observed at 10 muM. 2. Islet MAO activity was completely abolished in the presence of 10 muM of MAO inhibitors, while at lower concentrations, partial inhibition of enzyme could be achieved. A rise in the concentration of MAO inhibitors was accompanied by a decline in their potentiating effect on insulin secretion. No potentiation was observed at 1 mM concentration, and total inhibition of glucose-mediated insulin release was obtained in the presence of 5-10 mM of MA9 inhibitors. The potentiation of insulin release caused by two of the MAO inhibitors studied was abolished by addition of 1 muM epinephrine into the incubation medium. The basal rate of insulin release was insensitive to low concentrations of MAO inhibitors, but an inhibitory effect was obtained when concentrations were raised to 10 mM. In a comparative study, it was found that MAO activity was greater in liver than in islet tissue, while islets contained three times the activity of exocrine pancreas, consistent with previous findings in islet-cell tumors. The data presented here clearly show that MAO inhibitors are capable of both potentiating and inhibiting insulin release in vitro, depending on their concentrations. It is concluded that the stimulation of glucose-mediated insulin secretion may be related to the MAO inhibitory effects of the drugs, while the inhibition observed at concentrations greater than 1 times 10-3M is due to some other unidentified mechanism.


[Pyrazolic sulfanilamides. XIV. Hydroxyderivatives of 1-phenyl-5-sulfanilamidopyrazole and of 1-phenyl-3-methyl-5-sulfanilamidopyrazole]. A report is given of the variations in bacteriostatic activity on introduction of a hydrophilic group, the hydroxyl group (-OH), at positions 2',3' and 4' of the phenyl group linked to the heterocyclic nitrogen os 1-phenyl-5-sulfanilamidopyrazole (I: R = -H) and of 1-phenyl-3-methyl-5-sulfanilamidopyrazole (II: R = -H). The substances prepared for this purpose: 1-(hydroxyphenyl)-5-sulfanilamidopyrazoles (Ia)(Ib))(Ic)(-OH at 2', 3',4') and 1-(hydroxyphenyl)-3-methyl-5-sulfanilamidopyrazoles (IIa)(IIb)(IIc)(-OH at 2',3'4') in vitro tests of bacteriostatic activity against strains of S. aureus and E. coli gave the following results: See journal for results.


Ontogeny of mouse T-lymphocyte function. The development of lymphocytes within the fetal and neonatal BALB/c mouse thymus is reviewed with particular emphasis on the maturity of immunologic functions. Fetal thymocytes respond by vigorous proliferation to stimulation by allogeneic lymphoid cells or by phytohemagglutinin. Such reactivity is much diminished in neonatal thymus or thymic-derived (T) cells in neonatal spleen. Splenic T cells seem to mature more slowly than immunoglobulin-bearing B lymphocytes in the neonatal spleen, but the finding is confounded by the presence of large numbers of "suppressor" T cells in the neonatal spleen. For example, the in vitro antibody response to the T-independent antigen dinitrophenyl-lysine-Ficoll is optimal by 2 or 3 weeks of age, but the in vitro response to T-dependent sheep erythrocytes does not reach adult levels until 6 weeks of age, suggesting a deficiency in T "helper cells." The response of neonatal spleen cells to sheep erythrocytes cannot be reconstituted by adult T cells however, unless neonatal splenic T cells are first depleted by anti-Thy 1 serum and complement. The target of this T suppressor cell seems to be only B cells, and not other T cells. The overall sequence of T lymphocyte maturation in the mouse seems to start with large numbers of reactive T cells as well as some functionally active helper or effector T cells in early neonatal life, and finally to achieve a stable equilibrium between T cell subpopulations between 5 and 6 weeks of age.


Development of B lymphocytes-1,2.

Development of B lymphocytes-1,2. A rough outline of the development pathway of stem cells to B lymphocytes in mammals is beginning to emerge. The stem cell is most probably either identical to or closely related to the spleen colony-forming unit or hemopoietic stem cell. The stem cell appears not to be specificity-restricted and to lack Ig surface receptors. Whether part of B-cell development in mammals takes place in a specialized lymphoid environment, analogous to the bursa of Fabricius in birds, is still unresolved. The bone marrow does not form such an environment. The bone marrow and spleen, but not the lymph nodes, of mice contain a nondividing B-cell precursor population at an intermediate stage of development between the stem cell and B cell. This population has Ig surface receptors and is specificity-restricted. It cannot be initiated into differentiating into antibody-producing cells. However, under appropriate conditions it can give rise to functional B cells within 3 days. Newly-formed B cells (designated B1) are physically different from mature B cells (designated B2) and appear to be a dividing population of cells differentiating from the B-cell precursor to B2 nondividing cell populations.


Pre- and postnatal enzyme capacity for drug metabolite production. Most lipid-soluble foreign compounds including drugs, insecticides and many environmental pollutants are metabolized in animals by cytochrome P-450 enzymes in the endoplasmic reticulum of liver. These enzymes are virtually absent in fetuses of laboratory animals, but their activities increase to adult levels within 3-8 weeks after birth. In human fetuses, the enzymes appear during the first half of pregnancy, and their activities during gestation reach about one third of those found in adults. The species differences in fetal activities apparently parallel the differences in the development of liver endoplasmic reticulum. In laboratory animals, the rough-surfaced reticulum does not develop until 4 days before birth adn the smooth-surfaced retuculum develops only after birth. In man, however, the rough-surfaced form appears at about 7 to 9 weeks of gestation, whereas the smooth-surfaced form appears at about the 3rd month of pregnancy. Despite the early development of these enzymes in humans, they probably play only a minor role in limiting the accumulation of most foreign compounds in human fetuses. Nevertheless, they may play an important role in drug-induced toxicities, particularly those that are mediated through the formation of chemically reactive metabolites.


Indirect consequences of low-level use of antimicrobial agents in animal feeds. Antibiotics may be classified as "therapeutic" or "feed" depending on their use for treatment of disease or for promotion of growth in animals. The mechanism of growth promotion is largely unknown, but it has been shown that the specific feed additive causes defined lesions in the cell wall of Escherichia coli. This effect may sensitize the bacteria to the action of therapeutic drugs. Damage to the bacterial cell wall may also cause the bacteria to be more susceptible to the body defense mechanisms. One form of stress may be defined in terms of a disturbance of intestinal flora brought on by relocation, group mixing, change of diet, or travel. Prolonged feeding of antimicrobial agents to animals has not been shown to be responsible for the current high level of R factors in the intestinal tract of man.


Induction of timed ovulation with synthetic luteinizing hormone-releasing hormone in women undergoing insemination therapy. I. Effect of a single parenteral administration at midcycle. Synthetic LH-RH was administered by various routes to 19 patients during 37 treatment cycles in an attempt to trigger ovulation in association with insemination therapy. Thirty-five cycles were ovulatory; four pregnancies occurred, but two of them ended in spontaneous abortion. In 20 cycles, the hyperthermia response occurred one to three days after LH-RH administration, suggesting that ovulation occurred as a direct consequence of administered LH-RH. Low levels and delayed peak secretion of progesterone were found in two patients who had serial progesterone determinations during the luteal phase after LH-RH administration. Ovulation timing with LH-RH appears feasible, but efforts to ensure the availability of a mature follicle are required.


Use of synthetic luteinizing hormone-releasing hormone in treatment of oligospermic men: a preliminary report. Synthetic luteinizing hormone-releasing hormone (LH-RH) was administered to four normogonadotropic, oligospermic men (24 to 39 years of age) who had no endocrinologic, urologic, or associated vascular disease, to assess its possible therapeutic value in male infertility. Previous testicular biopsies of these subjects indicated alteration of spermiogenesis only. LH-RH (mean dose, 500 mug/day) was administered intramuscularly for 100 to 135 days. Each patient had at least two sperm count before starting therapy and had one every 20 to 30 days during and for two to five months after treatment. The sperm count, semen volume, sperm motility and morphology, and seminal plasma concentrations of fructose and citric acid were studied in each semen sample. In three of the four patients, urinary LH and FSH excretion and plasma testosterone levels were also measured. The sperm count increased clearly in two subjects 30 to 80 days after therapy started; the response was small in the third subject and negative in the fourth. The remaining parameters followed variable courses. Libido increased in all subjects. In the post-treatment period, the two patients who had shown the best response during treatment experienced a new and abrupt increase in the sperm count which remained well above initial values at the end of follow-up. LH-RH appears to be of value in the treatment of certain types of oligospermia, but several issues remain unsettled.


Unusual arrangement of agranular endoplasmic reticulum in the interstitial cells of the cat ovary. An unusual arrangement of agranular endoplasmic reticulum in the interstitial cells of the cat ovary is described. The structural unit of this arrangement in cross section consists of five membranous sacs (800 to 900 A long and 250 to 350 A wide) radially coverging toward a microtubule. The sacs are separated from each other by three angles of 60 degrees and two of 90 degrees. In longitudinal section, the sacs appear to be long parallel cisternae. The geometric, radial arrangement seems to arise from a gradual incurvation and convergence of parallel units in these membranes. The microtubules may act as guides for the polarization and resulting unusual arrangement of the membranes. The purpose of such polarization may be related to a change from protein to steriod synthesis in these cells.


History of resins in dentistry.

History of resins in dentistry. The period of 175 years from 1800 to 1975 represents one of significant advancement in prosthetic and restorative dental service. The transition from the time of ill-fitting dentures, fashioned from naturally occurring materials, to the application of synthetic resins for a long list of dental and surgical purposes described in this symposium represents a typical example of technical and professional advancement that has taken place throughout the world society during this same period. As noted in the beginning of this report, the advancement in dentistry has been possible through the cooperative efforts of contemporary scientists in many related fields. If denistry retains this good working relationship, as it is expected to do, then the advancements within the next two generations can significantly change and improve the practice of dentistry from what is presently known, by the application of additional new and modified dental resins.


Construction of denture bases from pour resins. Satisfactory dentures can be produced from pour resins at a considerable saving of processing time. The problems of porosity, voids, and heavy anterior tooth contact resulting from shrinkage of the resin during curing present difficulties which must be understood to be compensated for. The use of alginate and a recently introduced modified gypsum product as investing media, as well as centrifugation of the pour resin into the flask, offer no significant advantages over the originally introduced procedure. Further critical research will conceivably improve the pour resins and the corresponding processing techniques to the point that they can challenge heat-curing resins for the largest percentage of denture bases.


The acrylic jacket crown.

The acrylic jacket crown. An attempt has been made to cover briefly the many applications of the acrylic jacket crown. It is readily understandable that this type of restoration has many shortcomings but at the same time it has many useful and important applications in dentistry when properly employed. It is hoped that the specialist and generalist alike will have found some new and useful applications of the acrylic jacket crown.


Stimulation of glucagon secretion by ethanol-induced hypoglycemia in man. In fasted man, ethanol lowers plasma glucose by inhibiting gluconeogenesis with concomitant suppression of insulin release. Since glucose regulation of glucagon (IRG) secretion may be insulin dependent, we have evaluated IRG secretion in this setting of hypoglycemia and insulin deficiency. Mean IRG levels in six men fasted for fifty-six hours rose gradually from a basal level of 57 pg/ml. to 101 pg/ml. During the subsequent four-hour ethanol infusion, mean glucose concentration fell only 18 mg. per 100 ml. (26 per cent of pre-infusion values) yet IRG tripled to 265 pg/ml. Insulin (IRI) fell to unmeasurably low values. Alcohol given after only eight hours of fasting has no effect on plasma levels of glucose, IRG and IRI. These results suggest that the small decrease in extracellular glucose combined with relative insulin deficiency may cause inordinate intra-alpha cell glucopenia and result in exaggerated glucagon release.


Kinetics of taurocholate uptake by the perfused rat liver. The uptake of taurocholate (TC) by the perfused rat liver was studied by the multiple indicator dilution technique using single injections of 14-C-labeled TC and 99m-Tc-labeled albumin. For determination of the intra- and extravascular space, 51-Cr-labeled erythrocytes and albumin were used. TC was found to be distributed into an extravascular space equivalent to that of albumin. Analysis of the dilution curves, according to Goresky (Am J Physiol 207:13-26, 1964), revealed that the initial TC uptake rate increased with increasingly TC dose in a nonlinear fashion, exhibiting saturation kinetics which obeyed the Michaelis-Menten equation. The initial maximal uptake rats (Vmax) was found to be 32.5 nmoles per s and g of liver, a value exceeding the maximal steady state capacity (Tm) for excretion of TC into the bile. The half-saturation constant (Km) was 90.6 nmoles per g of liver, indicating that this system is operating far from saturation at physiological levels of bile acids in portal blood.


Gastrin cell hyperplasia in rats with chronic antral stimulation. Rats surgically prepared with the antrum transposed onto the colon were compared with suitable control rats, to investigate whether chronic antral stimulation modifies gastrin cell and other endocrine cell populations. Gastrin cell and argyrophil cell density per unit area were studied in antrum using a quantitative method after staining by immunofluorescence and Grimelius argyrophily, respectively. Both gastrin cells and argyrophil cells increased significantly in density per unit area after antrocolic transposition (P smaller than 0.001). The augmentation of gastrin cells per glandular tube (78%) was also significant (P smaller than 0.001). Electron-microscopic observations confirmed these results. On the other hand, in each group, the topographic distribution of the two categories of cells was different, and the number of gastrin cells was statistically greater than that of the argyrophil cells. Thus, it appeared evident that gastrin cells were not argyrophilic. Several hypotheses to explain the mechanism of this hyperplasia of endocrine cells are discussed. It is concluded that the increase in the gastrin cell density could be one possible explanation for the hypergastrinemia observed in the antrocolic transposition model.


Computer-aided diagnosis of lower gastrointestinal tract disorders. This paper reports a survey of human and computer-aided diagnosis in a prospective consecutive series of 301 patients admitted to the hospital with lower gastrointestinal tract disease. At initial outpatient contact (at which time endoscopy was customarily performed), the clinicians' diagnostic accuracy was 64.5%. After biopsy, radiology, and other investigative procedures, the clinicians' preoperative diagnostic accuracy rose t0 82.7%. When data from the house surgeon's case notes were fed into the computer, its diagnostic prediction proved accurate in 77% of the 301 patients, and when details of the outpatient endoscopy were added to the house surgeon's case notes, the computer's diagnostic accuracy rose to 84.7%. It is suggested that computer-aided analysis may have a limited part to play in discriminating between the common causes of lower gastrointestinal disorders in routine clinical practice.


Pathophysiology of Shigella diarrhea in the rhesus monkey: intestinal transport, morphological, and bacteriological studies. In contrast to the "toxigenic diarrheas" caused by Vibrio cholerae and Escherichia coli, the site and mechanism of fluid loss in shigellosis are unknown. The occurrence of watery diarrhea in shigellosis suggests involvement of the small bowel. Therefore, jejunal, ileal, and colonic water and electrolyte transport was studied in Shigella flexneri 2a-infected monkeys. Infected animals fell into three groups: dysentery alone, diarrhea alone, or diarrhea and dysentery. In controls, net water, sodium, and chloride absorption was seen in the jejunum, ileum, and colon. All infected animals demonstrated diminished colonic absorption or net colonic secretion. In monkeys with dysentery alone, this was the only transport defect observed. In contrast, animals with diarrhea either alone or in combination with dysentery, exhibited net jejunum secretion. Ileal transport was normal in all animals. A severe colitis with intramucosal shigellae was seen in all symptomatic animals. In the jejunum or ileum, however, morphological changes were minimal and bacterial invasion was not seen. Therefore, unlike the "toxigenic" diarrheas," shigellosis is both a small and large intestinal disease. Mucosal invasion of the colon is essential to the development of a morphological and transport defect. Dysentery results from a colonic transport defect, while diarrhea is secondary to jejunal secretion superimposed on the defect in colonic absorption.


Comparison of secretory and histological effects of shigella and cholera enterotoxins in rabbit jejunum. The purpose of this study was to compare the actions of shigella toxin and cholera toxin to determine if mucosal damage is a prerequisite for shigella toxin-induced secretion. The secretory response to maximal doses of cholera toxin and shigella toxin were compared. The maximal rate of secretion and the electrolyte and protein concentration of the intestinal fluid were similar for both toxins. On the other hand, the time of onset after exposure to the toxin was 105 min for shigella toxin and 15 to 30 min for cholera toxin. In addition, cholera toxin-induced secretion was associated with depletion of goblet cell mucus, whereas no change was seen in association with the response to shigella toxin. Other than goblet cell depletion, there were no histological differences between loops secreting in response to cholera toxin and to shigella toxin. Finally, the secretory effect of the toxins are not additive. These studies suggest that, in spite of apparent differences in the patterns of secretory response to the two toxins, they may share a rate-limiting step in the secretory process.


A study on the effect of certain compound during elimination of plasmids in Escherichia coli. Of several chemicals tested on the elimination of plasmids from Escherichia coli K-12, the compound designated ICR-170 was most effective, applied at 100 mu g/ml, the effect being comparable to that of acriflavin. It had no effect on the elimination of the R1 plasmid from Escherichia coli JC 5455.


DNA synthesis - dependent cell division of Escherichia coli 15 TAU after arginine and uracil starvation. Extensive cell division after synchronization of Escherichia coli 15 TAU by arginine and uracil starvation occurs only when DNA synthesis is permitted to proceed by at least a short pulse of thymine applied between 30 and 60 min after transfer of synchronized culture to thymine-free medium with arginine and uracil. The time schedule of synchronized cell division in dependence on the schedule of intervals of DNA synthesis and inhibition of DNA synthesis was determined. The termination of replication cycles which were not completed to the very end during arginine and uracil starvation seems to be the decisive event for subsequent cell division after synchronization.


Enhancement of Moloney sarcoma virus-induced tumor growth by concanavalin A. Adult mice injected intraperitoneally with concanavalin A (con-A) commencing with the onset of 6mm. Moloney sarcoma virus (MSV)-induced tumors showed a marked enhancement of tumor growth compared to untreated control mice; whereas, treatment with con-A beginning on day 0 relative to MSV inoculation had no effect. Antibody production as measured by indirect membrane fluorescence was not affected by con-A. Since in vivo administered con-A has a specific suppressive effect on T-lymphocytes it is suggested that the primary immunologic response to MSV antigens is a function of T-cells.


Comparative mitogenic responses of T-cells and B-cells in spleens of mice of varying age. Mouse spleen cell mitogenic responses to Phytohemagglutinin (PHA), conconavalin A (con A), staphylococcal enterotoxin B (SEB) and lipopolysaccharide (LPS) were studied as a function of age. In CD-1 mice, spleen cell DNA synthesis in response to stimulation by PHA, con A and SEB varied from 18% - 33% of adult values for the first 3 weeks of life. By 6 weeks, the PHA response was 63% of the adult response and achieved adult values by 8 weeks of life. In contrast to these T-cell responses, DNA spleen cell synthesis in response to LPS was approximately that of adults as early as 7 days after birth. When spleen cell mitogenic responses were studied in C57B1/6J mice a similar pattern emerged. C57 spleen cell responses to PHA, con A and SEB varied from 24% - 54% of adult values for the first 3 weeks of life. By 4 weeks, spleen cell responses to PHA and con A were approximately those of adults. Again, C57 spleen cell responses to LPS were equivalent to adult values as early as 7 days after birth.


The bursa dependence of chicken thymus-derived lymphocyte surface immunoglobulin. Thirty-three plus or minus 5 per cent of chicken blood lymphocytes were positively labelled by radioiodinated rabbit anti-chicken light chain antibody within 7 days of exposure of autoradiographs. After thymectomy, 93 plus or minus 1 per cent of lymphocytes were positive: conversely, following bursectomy only 5 plus or minus 3 per cent of cells showed positive labelling and then at a very low level. Hence, under these conditions of short-term exposure autoradiography, only B cells carried readily demonstrable immunoglobulin. If the sensitivity of the technique was increased by long-term exposure of the autoradiographs, immunoglobulin was demonstrable on all blood lymphocytes, including the T cells. In addition, 39 plus or minus 6 per cent of thymocytes were shown to carry immunoglobulin by the more sensitive technique. In both cases, however, this T-lymphocyte surface immunoglobulin was reduced, and in some cases absent, after bursectomy. At no stage, with either technique, did the grain count on the erythrocytes reach the control level of lymphocytes treated with 125I-labelled anti-keyhole limpet haemocyanin antibody. It seems likely that T-cell immunoglobulin is not an endogenously synthesized product but is acquired indirectly from the B lymphocytes.


A method for the separation of lymphocytes and plasma cells from the human palatine tonsil using sedimentation in an isokinetic gradient of Ficoll in tissue culture medium. Several methods for the dissociation of human tonsils into cell suspensions were compared. Dissociation of tonsils using 0-25 per cent trypsin gave both the largest number of total cells and the largest number of plasma cells per gram of tonsil. Lymphocytes and plasma cells were separated in a previously described isokinetic gradient of Ficoll in tissue culture medium. In the purest gradient fractions, lymphocytes were 97-2 plus or minus 1-9 per cent of nucleated cells. The purest gradient fractions contained 43-1 plus or minus 5-9 per cent plasma cells. More than 95 per cent of purified lymphocytes and plasma cells excluded Trypan Blue.


Proportional absorption. A method for determination of the relative specificity of antisera prepared against cells. Antisera prepared against a complex of antigens such as a tissue cell may produce a mixture of antibodies of different specificities, affinities and types. Proportional absorption permits determination of the comparative specificity of such antisera. It is performed by absorbing the antisera with an amount of  absorbent proportional to the initial content of (usually undesired) antibody to this absorbent; the potencies of desired and undesired antibodies are then separately determined. The method has been used to determine the specificity of a conventionally raised rabbit anti-mouse leukaemia serum, relative to one prepared with leukaemia cells admixed with rabbit antiserum against normal mouse lymphocytes (to block normal antigen sites on the leukaemia cell inoculum). The latter antiserum was cells as compared to normal splenic lymphocytes.


The effect of various immunosuppressive agents on mouse peritoneal macrophages and on the in vitro phagocytosis of Escherichia coli O4:K3:H5 and degradation of 125I-labelled HSA-antibody complexes by these cells. Large doses of hydrocortisone, cyclophosphamide, and methotrexate injected subcutaneously, and whole-body irradiation (500 rads) caused a reduction in the number of peritoneal cells (PE cells) obtained after intraperitoneal injection of the treated mice with proteose-peptone. The same dose of cyclophosphamide and irradiation induced morphological changes in PE macrophages. There were more giant cells in the peritoneal exudates from treated mice as compared to control mice. 'Pharmacological' and larger doses of hydrocortisone, methotrexate and azathioprine or anti-lymphocyte globulin had no effect on the in vitro phagocytic capacity of proteose-peptone-stimulated mouse PE macrophages. This also applied to doses of up to 50 mg/kg of cyclophosphamide. In contrast, whole-body irradiation (500 rad) and 100 mg/kg of cyclophosphamide decreased the phagocytic capacity of mouse macrophages in vitro and reduced the ability of PE cells to degrade 125I-labelled HSA-antibody complexes in vitro. The greatest effect was noted 4-5 days after whole-body irradiation or four to five subcutaneous injections of cyclophosphamide.


Inhibition of Escherichia coli by bovine colostrum and post-colostral milk. I. Complement-mediated bactericidal activity of antibodies to a serum susceptible strain of E. coli of the serotype O 111. Bovine colostral whey diulted in unheated bovine milk or in Kolmer saline containing 5 per cent pre-colostral calf serum was bactericidal for Escherichia coli NCTC 9703 (serotype O 111). Whey diluted in saline without pre-colostral calf serum was inactive. Bactericidal activity was abolished by heating at 56 degrees for 30 minutes or by pre-incubating with N-acetyl-L-tyrosine ethylester, indicating that complement present in milk or pre-colostral calf serum was involved in the bactericidal activity. Undiluted colostral whey was inactive even in the presence of up to 20 per cent pre-colostral calf serum, probably due to a prozone effect. Milk heated at 80 degrees for 10 minutes was also shown to have a limited anti-complementary effect. Post-colostral milk from some cows had bactericidal activity, whereas similar milks from other cows were either inactive or active only after addition of pre-colostral calf serum, indicating that antibodies to E. coli or complement or both may or may not be present in the post-colostral milk of different cows. The implications of these findings in relation to the bacterial flora of the intestinal tract of neonates are discussed.


Inhibition of Escherichia coli by bovine colostrum and post-colostral milk. II. The bacteriostatic effect of lactoferrin on a serum susceptible and serum resistant strain of E. coli. Two strains of Escherichia coli were inhibited by complement-inactivated cow serum and to a lesser extent by precolostral calf serum devoid of specific antibodies. They were not inhibited by undiluted colostral whey or milk but colostral whey became bacteriostatic after dialysis or dilution in Kolmer saline and addition of precolostral calf serum or lactoferrin. The inhibition in all these fluids was due to iron-binding proteins (transferrin or lactoferrin). Undiluted dialysed milk was not inhibitory because of its low content of lactoferrin but became inhibitory after addition of 1 mg/ml of lactoferrin. The lack of inhibition in undiluted whey is due to the high concentration of citrate in colostral whey (and milk) and it is suggested that citrate competes with the iron-binding proteins for iron and makes it availabe to the bacteria. Addition of bicarbonate, which is required for the binding of iron by transferrin and lactoferrin, can overcome the effect of citrate; hence, the bacteriostatic effect of cow serum and precolostral calf serum is due to the presence of both transferrin and bicarbonate as well as the low lefel of citrate.


Suppression of extablished Friend virus leukemia by statolon: potentiation of statolon's leukemosuppressive activity by chlorite-oxidized oxyamylose. Treatment of Friend virus (FV)-infected mice, 3 days after FV inoculation, with statolon, an extract of the mold Penicillium stoloniferum, induces interferon and restores immunocompetence to viral and nonviral antigens such as sheep erythrocytes. Clinical remissions are established in 20 to 70% of the infected mice. Cholorite-oxidized oxyamylose administered intraperitoneally 24 h before, 3 h before, or 3 h after statolon enhanced interferon production, but the increased number of mice protected against FV disease was more closely related to the associated enhanced synthesis of FV cytotoxic antibody. The prolonged selective immunodepression to intraperitoneal sheep erythrocytes after intraperitoneal administration of chlorite-oxidized oxyamylose-statolon appeared to be related to a stimulation in number and erythrocyte-phagocytic capacity of peritoneal macrophages. The marked activation of macrophages in FV leukemic mice after such treatment may also have contributed to the enhanced FV leukemosuppressive effects of chlorite-oxidized oxyamylose-statolon.


Immunoelectron microscopic identification and localization of Streptococcus sanguis with peroxidase-labeled antibody: localization of surface antigens in pure cultures. An indirect method of localizing antigens with horseradish peroxidase-labeled antibody was used to identify and localize surface antigens of Streptococcus sanguis at the ultrastructural level. An electron dense layer surrounding the cell wall could be distinguished without any additional electron microscope staining. This labeled layer represents an immune complex consisting of bacterial surface antigens, specific rabbit antisera, and peroxidase-labeled goat anti-rabbit globulins. Although with undiluted antisera slight cross-reactions occurred with S. salivarius and S. miteor (mitis), these could be readily distinguished from the more intense homologous reaction by their patchiness and the difference in distribution of the label. These cross-reactions were eliminated by appropriate dilutions of the antiserum. No cross-reactions occurred with S. mutans, S. faecalis, Actinomyces species, or Bacterionema, microorganisms wents indicated that horseradish peroxidase can become non-specifically adsorbed to the membrane of certain bacterial cells. Appropriate controls must, therefore, be included for localization of membrane associated antigens with horseradish peroxidase.


Regional immunosuppression induced by Plasmodium berghei yoelii infection in mice. Plasmodium berghei yoelii infection in mice severely depressed the splenic antibody response to sheep erythrocytes but had lettle effect on antibody formation in lymph nodes.