Renal biogenesis of erythropoietin.

Renal biogenesis of erythropoietin. The widespread and ever expanding use of dialysis in the maintenance of patients with chronic renal disease has added an urgency to the study of the biogenesis of erythropoietin. It seems almost certain that erythropoietin could ameliorate, if not eliminate, the anemia of uremia, but unfortunately, erythropoietin is still not available in therapeutic quantities. Initially, erythropietin was though to be produced by the kidneys" but then the attention became directed at the liver. It was proposed that erythropoietin was produced there as an inactive precursor and that the kidney only acted as an oxygen sensor and as a producer of an erythropoietin-activating enzyme. Recent studies summarized here show that an isolated perfused kidney in the absence of any extrarenal substrate or precursor can synthesize erythropoietin. Consequently, it appears almost certain that the kidney is the endocrine organ of origin of erythropoietin. Further studies suggest that erythropoietin formation involves a phase of oxygen sensing and programming and a phase of synthesis. These phases probably occur in the same cell, and the renal cortex appears to be the most likely location for such cells. The current inability to extract erythropoietin from kidney homogenates is discussed but, unfortunately, not adequately explained.


Calcium metabolism in renal failure.

Calcium metabolism in renal failure. Osteodystrophy is almost universally present in chronic renal failure. Mild, but detectable, abnormalities--especially in parathyroid hormone (PTH) secretion--occur even when the glomerular filtration rate is greater than 30 cc/min. Osteomalacia is common in areas in which vitamin D intake and exposure to sunlight are minimal; when these factors are plentiful, osteitis fibrosa predominates. Osteoporosis is seen with increasing frequency in hemodialyzed patients. Nonosseous complications of secondary hyper-parathyroidism include hypercalcemia, metastatic calcification and pruritus. The most important factor in the medical therapy of osteodystrophy is control of serum phosphate levels. Next, a positive calcium balance must be provided either by giving vitamin D as dihyrdotachysterol, raising dialysate calcium or administering calcium orally. Parathyroidectomy is sometimes indicated, especially when the patients are transplant candidates and manifest hypercalcemia. Whether or not transplant is contemplated, patients with persistently high calcium-phosphate products, severe metastatic calcification or rapidly progressive osteodystrophy should be considered for parathyroidectomy. Newer, experimental vitamin D preparations, such as 1,25-dihydroxycholecalciferol or 1-alpha-hydroxycholecalciferol, should improve the managemet of patients with renal osteodystrophy and decrease the need for parathyroidectomies.


Renal handling of low molecular weight proteins. The renal filtration, absorption and final disposal of lysozyme (lysozyme--mol wt 14,000), insulin and growth hormone were studied to gain a better quantitative understanding of the fundamental variables involved in the renal handling of low molecular weight proteins. The glomerular barrier offers little hindrance to the filtration of lysozyme, the glomerular sieving coefficient being 0.8 plus or minus 0.1 (SD). The intrarenal route by which injected lysozyme accumulates in the kidney is via filtration and subsequent absorption (uptake) by renal tubular cells. Uptake or adsorption from the peritubular side is negligible compared to luminal uptake. Renal clearance and renal titration experiments in the intact dog and in the isolated perfused rat kidney showed that the lysozyme absorption process can be best characterized as high capacity, low affinity transport system which is directly or indirectly dependent on energy input. The final disposal of absorbed 125I-lysozyme, 125I-insulin and 125I-growth hormone was studied in the isolated perfused rat kidney by measuring the radioactivity by gel chromatography. The rate of release of radioactivity as well as its nature was dependent on the molecular species of the absorbed protein. The rate of release was higher for 125I-insulin and 125I-growth hormone and lower for 125I-lysozyme. Lysozyme absorbed from the luminal side was released to the perfusate both as intact protein molecules and as catabolic products, whereas absorbed 125I-insulin was almost entirely released to the perfusate as catabolic products. It is concluded that low molecular weight proteins are extensively filtered by the kidney, absorbed from the luminal side by renal tubular cells and released back to the circulation either as intact molecules or as catabolic products (amino acids and polypeptides). This process contributes in an important way to the plasma turn-over of low molecular weight proteins including peptides and proteins hormones.


Growth in children with renal failure.

Growth in children with renal failure. Chronic renal insufficiency in childhood is often associated with growth delay. Undernutrition, osteodystrophy and diminished somatomedin levels appear to be importantly related to this growth delay. In many but not all children with renal failure and growth retardation, somatomedin levels are diminished, growth hormone levels are normal or elevated, and glucose tolerance is often abnormal. Following renal transplantation, growth restoration appears to be related to bone age, renal function, somatomedin and steroid therapy.


Clinical effects of bilateral nephrectomy. The effects of removal of all renal tissue on hematopoiesis, osteodystrophy, blood pressure regulation and metabolic functions are reviewed; and, the indications for, and results of, bilateral nephrectomy are discussed. Nephrectomy results in a more severe anemia in dialysis patients which is poorly responsive to androgen therapy. No differences were detected in the severity of osteodystrophy between nephric and anephric patients. However, bilateral nephrectomy can occasionally result in the acute onset of hypocalcemia. Blood pressure regulation must be accomplished in the absence of a functioning renin-angiotensin system. This is largely on the basis of volume, but changes in vascular tone may also be significant. Little is known about the metabolic consequences of nephrectomies. The effect on substances metabolized by the kidney is an area for further investigation. Kidney tissue should be preserved, if at all possible, and nephrectomy performed only for specific indications.


Hereditary complement (C2) deficiency with dermatomyositis. A 60 year old white man in previous good health presented with a 6 month history of progressive muscle weakness. Clinical and laboratory findings were typical of dermatomyositis. Muscle biopsy confirmed the presence of inflammatory myopathy; deposits of immunoglobulin G (IgG), immunoglobulin M (IgM) or third component of complement (C3) were not detected by immunofluorescence. No evidence was found for an associated neoplasm. An unexpected finding was the total absence of serum hemolytic complement activity. Further investigation revealed that the complement defect was attributable to a selective and total absence of the second component of complement (C2), as determined by both functional and immunoprecipitin assays. Family studies indicated that the defect was inherited in an autosomal recessive manner, as has been observed in the previously reported C2-deficient kindreds. This case demonstrates that typical muscle lesions of dermatomyositis can occur in the presence of a complement defect which would preclude activation of the classic (C1-C4-C2) complement pathway. The case is of further interest as one of a series of recently reported associations of rheumatic diseases with hereditary complement deficiencies. Study of the functional properties of the propositus' C2-deficient serum demonstrated normal generation of chemotactic activity in the presence of endotoxin or aggregated IgG, and normal or near normal bactericidal activity against Salmonella typhi O 901 and Hemophilus influenzae, type b. These findings emphasize the importance of the alternate (properdin) pathway of complement activiation in these functions.


Lymphocyte surface characteristics in malignant lymphoma. The surface of lymphocytes obtained from fresh biopsy specimens from 41 patients with malignant lymphoma and from 30 normal subjects or patients with non-neoplastic lymphadenopathy were investigated. Immunoglobulin on the cell surface was used to identify B cells, whereas T cells were recognized by their reactivity with an antithymocyte antiserum and their ability to form rosettes with sheep erythrocytes. Normal and inflammatory lymph nodes were composed predominantly of T lymphocytes, as were nodes from 14 patients with Hodgkin's disease. Two thymomas were T cell proliferations, whereas a node from a patient with ataxia-telangiectasia was devoid of T lymphocytes. The presence of immunoglobulin on the cell surface indicated that 19 of 21 lymphocytic lymphomas were B cell proliferations, whereas the cells from 3 histiocytic lymphomas (reticulum cell sarcomas) and 1 mixed histiocytic and lymphocytic lymphoma were devoid of surface immunoglobulin. In immunoglobulin-positive tumors, one predominant heavy chain and one predominant light chain could usually be identified, thus establishing the clonal character of the neoplastic proliferation. Ten of 11 diffuse poorly differentiated lymphocytic lymphomas were composed of cells with large amounts of surface immunoglobulin, whereas only 1 of 5 diffuse well differentiated lymphocytic tumors contained such abundant surface immunoglobulin. The surface immunoglobulin data indicate the existence of at least two subspecies of B cell neoplasms. A small lymphocyte with sparse surface immunoglobulin proliferates as diffuse well differentiated lymphocytic lymphoma and chronic lymphocytic leukemia, whereas a larger lymphocyte with abundant surface immunoglobulin proliferates as diffuse poorly differentiated lymphocytic lymphoma and lymphosarcoma cell leukemia.


Hodgkin's disease and acute leukemia. Report of eight cases and review of the literature. Eight cases of Hodgkin's disease and acute leukemia are reported. An additional 74 cases of acute myelocytic leukemia or one of its variants, 11 cases of acute lymphocytic leukemia, 12 cases of chronic myelocytic leukemia and 37 cases of chronic lymphocytic leukemia associated with Hodgkin's disease are reviewed from the literature. In 3 of the 82 patients with acute myelocytic leukemia and Hodgkin's disease, the two diseases occurred simultaneously. Of the remaining 79 patients, 76 had received radiation therapy for their Hodgkin's disease and acute myelocytic leukemia had developed 1.2 to 19 years later (mean 6.5 years). Thirty-four of these patients also received antineoplastic chemotherapy. Only three patients with Hodgkin's disease were treated with multiple chemotherapy alone; in these, Hodgkin's disease developed 1.2, 1.5 and 3.2 years later. In 4 of 11 patients with acute lymphocytic leukemia and Hodgkin's disease, the two disorders occurred simultaneously. The other seven patients were all treated with radiation for their Hodgkin's disease, and acute lymphocytic leukemia developed 2 to 8 years later (mean 4.5 years). Three of the 7 patients also received alkylating agents. It is concluded that the development of acute leukemia, mostly acute myelocytic leukemia but also acute lymphocytic leukemia, during the course of Hodgkin's disease, is most likely related to radiation therapy. There is as yet insufficient evidence to implicate intensive chemotherapy in the causation of acute leukemia since in only three patients with Hodgkin's disease treated with chemotherapy alone has the development of acute leukemia been reported. It is possible, however, that chemotherapy potentiates the effect of radiotherapy. 2t is also possible that acute leukemia is part of the natural history of Hodgkin's disease and is occurring with greater frequency because of improved survival in Hodgkin's disease since the introduction of better radiotherapeutic and chemotherapeutic treatment regimens.


Clinical experience with transfusion of granulocytes obtained by continuous flow filtration leukopheresis. Granulocytes obtained by continuous flow filtration leukopheresis (CFFL) were transfused to 21 patients on 131 occasions. An average of 28.2 times 10-9 granulocytes were administered per transfusion. These cells were more than 90 per cent viable by dye exclusion, ingested latex particles normally and had almost normal bactericidal activity. Migration to skin windows was demonstrated on four of six attempts, but 51-Cr-labeling studies failed to show localization in infected areas on six occasions. Post-transfusion granulocyte count increments averaged 225/mul and were transient. Significant transfusion reactions occurred during 35 transfusions to 13 patients. Reactions occurred in some patients without demonstrable alloimmunization and after six infusions of HL-A identical or compatible cells. Definite clinical improvement was noted in three recipients. Stabilization of infection with patient survival occurred nine times, and progression of infection with death eight times. Granulocytes obtained by CFFL are viable and functional. Their transfusion is not without risk and must still be considered an investigative procedure of suggestive but as yet unproved clinical efficacy.


Drug-induced immune hemolytic anemia.

Drug-induced immune hemolytic anemia. Drug administration causes from 16 to 18 per cent of cases of acquired immune hemolytic anemia. The pathogenesis of erythrocyte sensitization by drug-related antibody with or without fixation of complement is variable, and there is a relationship between the responsible drug, the mechanism of red cell sensitization, clinical manifestations and laboratory methods of diagnosis. Drugs such as phenacetin and quinidine form a complex with the antidrug antibody, and the immune complex attaches to red cells usually fixing complement and causing acute intravascular hemolysis. Other drugs (e.g., penicillins), when given in large doses, coat normal red cells in vivo and in some patients a high titer IgG anti-drug antibody develops which reacts with the coated cells. Hemolytic anemia may develop with red cell destruction being primarily extravascular. Cephalosporins cause positive direct antiglobulin tests in a small percentage of patients either by the same mechanism as penicillins or by modification of the red cell membrane leading to nonimmunologic absorption of serum proteins. Hemolytic anemia has been reported only rarely. A few drugs (notably alpha methyldopa) cause the development of autoimmune hemolytic anemia. Knowledge of clinical manifestations and laboratory aids to diagnosis is necessary to distinguish immunohematologic abnormalities caused by drugs from other causes.


Syndrome and pancreatic disease, subcutaneous fat necrosis and polyserositis. Case report and review of literature. Immunologic evaluation of a patient with pancreatitis, subcutaneous fat necrosis, pleuritis, pericarditis and synovitis is presented. The previously recognized syndrome of pancreatic disease, subcutaneous fat necrosis and arthritis is reviewed. Based on analysis of all the cases described in the English language literature it is suggested that this syndrome be expanded to include polyserositis rather than arthritis alone. Although experimental and clinical evidence tends to implicate physiocochemical tissue injury by pancreatic lipase as the primary pathogenic mechanism in this syndrome, studies in our patient suggest the possible contribution of immune-mediated injury. Supporting data include eosinophilia, biopsy demonstration of vasculitis antedating the subcutaneous fat necrosis, immunofluorescent identification of immunoglobulin G (IgG) and C3 in the pleura, and reduced levels of total hemolytic complement in the serum, and pleural and pericardial effusions.


Perturbations of humoral and cellular immunity in a patient with pigmented villonodular synovitis. Several parameters of the immune system have been studied in a patient with pigmented villonodular synovitis. Numerous immunoglobulin-synthesizing cells were found by immunofluorescent technics in both synovium and synovial fluid. Phytohemagglutinin-induced in vitro blastogenesis of peripheral blood T lymphocytes was absent whereas B-lymphocyte blastogenesis was preserved. These results indicate a necessity for evaluating immune responsiveness in other patients with pigmented villonodular synovitis.


Determining antibiotic levels in the blood. Methods for the rapid determination of antibiotic blood levels require particular organisms or special conditions or long incubation. A method was developed for the rapid determination of antibiotic blood levels which requires no special equipment and is rapid. Special stains are not required. Utilizing bacteria from standard Kirby-Bauer sensitivity plates, a suspension is prepared and a pour plate made. Organisms which are sensitive to the questioned antibiotic and resistant to other antibiotics the patient is receiving are chosen. Muller-Hinton is the best general agar. Wells are cut, and antibiotic standards, antibiotic controls, and the patient's serum introduced. Within three to four hours, zone sizes can be read and the antibiotic blood level calculated. The materials and technical facility required should not preclude any laboratory from successfully performing the method.


Adaptive Behavior Scale as a screening measure for special-education placement. The utiltiy of the Adaptive Behavior Scale in the special-education placement of 126 institutionalized mentally retarded children and adolescents was examined. An investigation of relationships among Adaptive Behavior Scale (Part 1) scores, Wide Range Achievement Test scores, and IQ scores revealed a significant relationship between Adaptive Behavior Scale scores as well as between IQ scores and achievement test scores. Adaptive Behavior Scale and IQ were the most effective scores in providing significant discrimination between the special-education training levels. Adaptive Behavior Scale subdomain percentage scores were discussed as criteria for a computer-assisted placement of mentally retarded students in a special-education program.


Macrophage migration inhibitory factor in female sterility. Cellular sensitization against spermatozoa as antigen was studied in 21 fertile and 41 infertile female subjects. In addition, 20 virgin girls and a group of 11 women presenting positive spermagglutinating (in equal 8) and immobilizing (n equal 3) antibodies were investigated. Cellular sensitization was expressed in terms of macrophage migration inhibitory activity (MIF-reaction). For this screening program washed and pooled spermatozoa (WPS) and sonicated spermatozoa solutions (sss) were used as antigens. In 52 to 72 per cent of all cases, including primary and secondary sterility of unexplained cause, a positive MIF-reaction was encountered. Women with humoral sperm antibodies always revealed a cellular sensitization. Only the group of virgin girls demonstrated in no case a positive MIF-reaction. The need for the identification of the specific fertility-diminishing antigens is stressed as the results indicate a cellular sensitization in all cases that had sperm contact.


In vitro biosynthesis of TSH- and LH-releasing factors by the human placenta. Human chorionic membrane and human placental tissue contain substances with the same elution volume as hypothalamic TRF and LRF standards during separation by carboxyl-methyl-cellulose ion-exchange chromatography. Homogenates prepared from fresh human placentas delivered at term caused release of TSH (thyroid-stimulating hormone) and LH (luteinizing hormone) when bioassayed in rats. After dialysis of placental homogentes, synthesis of biologically active TSH-and LH-releasing factors can be shown to occur in vitro following the addition of appropriate precursor amino acids and cofactors.


Oral contraceptives and vitamins: a review. The literature concerning the influence of estrogen-containing oral contraceptives on vitamins is reviewed. The appearance of an elevated plasma concentration of vitamin A is probably without clinical importance, while there seems to be a clear connection between disturbances of vitamin B6 metabolism and mental symptoms. Low levels of folic acid and vitamin B12 have also been noted. Despite this, it is very rare that hematologic abnormalities develop during hormonal contraception. A reduced concentration of vitamin C in plasma and blood corpuscles has been reported. The clinical significance of these alterations is unknown.


Bacteremia on an obstetric-gynecologic service. In a one-year evaluation of the Obstetric-Gynecologic Services of the Los Angeles County University of Southern California Medical Center, bacteremia was confirmed on 144 occasions in 139 patients. This represented an over-all incidence of 7/1,000 admissions with gram-negative bacteremia observed in 3.1/1,000 admissions. There were four deaths in this series. The most frequently recovered aerobes were Escherichia coli, enterococci, and beta hemolytic streptococci, not Group A or D, while the most commonly isolated anaerobes were peptostreptococci, peptococci, and Bacteroides. These patterns of bacterial isolation should be acknowledged in antibiotic strategies for septic patients. There is a positive correlation between the incidence of intrapartum maternal and fetal monitoring and postpartum maternal bacteremia. The oncology patients were the most seriously ill women with bacteremia.


Pituitary gonadotropin responses to synthetic luteinizing hormone-releasing hormone in patients with typical and atypical polycystic ovary disease. Synthetic luteinizing hormone-releasing hormone (LH-RH) was administered intravenously to 17 women with polycystic ovary disease (PPCO), 16 women with hypothalamic amenorrhea (HA), and nine women with normal cycles. The serum levels of follicle-stimulating hormone (FHS) and luteinizing hormone (LH) were measured at frequent intervals before and after LH-RH injection. The PCO patients were arbitrarily divided into two groups on the basis of ovarian morphology. The eight patients with large, "typical" PCO's (Type I) showed a greater LH response and a lower FSH response to LH-RH than did the nine patients with smaller, "atypical" PCO's (Type II). The LH response in Type I co was also greater than that in HA, but the FSH response in both types of PCO was significantly less than that in HA. The gonadotropin responses to LH-RH in Type I PCO were quite similar to those occurring in normal women during the two to three days prededing ovulation. The results suggest that excessive LH secretion and/or impaired FSH secretion may be etiologically significant in PCO but do not clearly differentiate whether the stimulus for this pattern of gonadortopin secretion occurs at the hypothatamic or pituitary level.


The present status of prenatal detection of neural tube defects. In experimentally induced myelocele in rats, efforts to find neural cells in amniotic fluid (AF) were unsuccessful. Creatine phosphokinase (CPK) and aldolase concentrations studied in serum of 118 and cerebrospinal fluid (CSF) in 9 patients with myelomeningocele showed serum CPK to be significantly elevated and more responsive to additional muscle injury than aldolase, but both enzymes appeared in lower concentrations in patients with myelomeningocele than those with infantile atrophy or cerebral palsy. In CSF, CPK, and aldolase concentrations averaged 4.2 I.U. and 2.7 S.L.U. per milliliter, respectively. Significant CPK elevation (p less than 0.001) was also found in AF from myeloschitic fetuses and maternal rat serum. Although these findings suggest that increased CPK concentration is an indicator of myelocele in rats, the technique is impractical for prenatal detection of human fetus occurs too late in gestation. This does not, however, preclude the value of CPK for detecting onset of paraparesis. In all myeloschitic human fetuses, the CSF communicates directly with AF for at least 3 to 4 weeks. This implies that CSF is probably the principal source of increased alpha-fetoprotein concentration encountered in AF of all pregnancies with NTD. When biological variables are recognized, it is evident that increased concentration of amniotic fluid alpha fetoprotein is a reliable indicator of fetuses with open myelocele and/or anenciphalus.


Volume and sodium concentration studies in 300 saline-induced abortions. In an attempt to clarify the role of volume changes and Na concentration in saline-induced midtrimester abortions, 800 amniotic fluid samples were studied in 300 cases. The immediate postinfusion values of Na ranged as high as 3,000 mEg. per liter and as low as 1,200 mEg. per liter, still resulting in abortion. In earlier gestation, higher Na concentration and greater volume increase was needed for successful induction of abortion. Volume increase, within limits, compensated for Na concentration decrease. Osmotically induced volume changes were proportionatenate to the advancing stage of gestation.


Infections with mycoplasma and bacteria in induced midtrimester abortion and fetal loss. Placentas and fetuses from cases of induced midtrimester abortion and fetal loss (13 weeks of pregnancy or more) were examined microbiologically. In the series of midtrimester fetal loss, bacteria were isolated in 14 per cent and genital Mycoplasma in 37 per cent of the placentas. In induced abortion, 18 per cent of the placentas grew bacteria and in only one case out of 27 was a Mycoplasma strain isolated. Fetal organs yielded cultures positive for bacteria in 16 per cent and for genital Mycoplasma in 23 per cent of the spontaneous fetal loss material, whereas pyogenic cocci were isolated from fetal organs in 18 per cent and Mycoplasma in no case of induced abortion. In the positive cases, the fetal lungs were most often infected (in 34 out of 37 positive cases of spontaneous and 4 out of 5 positive cases of induced abortion). On the other hand, microorganisms could be isolated from the brain in only 5 out of 28 cases of spontaneous and none of induced abortion. In the series of fetal loss, positive microbiologic findings from the fetal organs seemed correlated to prolonged bleeding before abortion or labor and to premature rupture of membranes.


Evaluation of Deladumone OB in the suppression of postpartum lactation. Of 235 patients receiving Deladumone OB, more than 39 per cent complained of breast pain or swelling; 83 per cent were pleased with the medication. Of 215 patients receiving a placebo, more than 82 per cent complained of breast pain or swelling; 67.5 per cent were pleased with the medication.


Calcium metabolism in pregnancy: a review. Calcium metabolism in pregnancy is a complex process involving calcium, phosphorus, vitamin D, parathyroid hormone (PTH), and calcitonin (CT). Calcium absorption is enhanced in pregnancy, and increased storage in the maternal skeleton probably occurs as well. Adequate amounts are provided by the current Recommended Dietary Allowance of 1,200 mg. daily which can be met readily by natural foods, specifically milk. If supplemental calcium is given, a nonphosphate salt is probably advisable, since some evidence suggests that excessive phosphate intake may be related to leg cramps in pregnancy. Vitamin D is necessary for optimal calcium utilization in pregnancy, although the possibility of fetal toxicity with overdosage has been suggested. From a review of available information with respect to maternal-perinatal calcium interrelationships, I propose the following hypothesis: While total maternal serum calcium declines during pregnancy because of the physiologic hypoalbuminemia, the level of ionic calcium remains constant, in part, at least, because of increasing maternal PTH output. The placenta plays a primary role in fetal calcium metabolism by transporting calcium ions from the mother to the fetus against a concentration gradient. Relatively high fetal ionic calcium levels cause suppression of PTH and stimulation of CT in the fetus, facilitating growth of the fetal skeleton. With sudden loss of the placental source of calcium at birth, the newborn infant becomes functionally hypoparathyroid and/or hypercalcitonemic, and the serum calcium level declines until 3 to 4 days of life when PTH rises and CT falls with a resultant slight rise in calcium.


Puerperal endometritis: a prospective microbiologic study. This study was undertaken to determine the flora in the endometrial cavity of two groups of puerperal patients: 47 afebrile control subjects and 27 with endometritis. A transcervical culturing technique designed to avoid cervical contamination and to preserve aerobes, anaerobes, and mycoplasma was employed, and bacterial colony counts were reported semiquantitatively. No difference was found in the flora of these two groups. Furthermore, the most common pathogenic organisms (in decreasing order of frequency) were peptostreptococci, peptococci, Bacteroides sp., gamma-streptococci, enterococci, beta-streptococci, alpha-streptococci, and E. coli.


Sperm-agglutinating antibodies and testicular morphology in fifty-nine men with azoospermia or cryptozoospermia. The relationship between the state of the germinal epithelium and the type and titer of circulating sperm-agglutinating antibodies has been investigated in a series of 59 azoospermic or occasionally cryptozoospermic men. The patients were grouped according to the condition of the germinal epithelium as observed from testicular biopsy specimens, as well as to type and titer of circulating sperm-agglutinating antibodies investigated by a previously described microagglutination technique. Evidence is presented to suggest that the presence of mature spermatozoa in the testicular structures may be a prerequisite for the spontaneous production of circulating sperm-agglutinating antibodies, at least of the head-to-tail (H-T) agglutinating type. Furthermore, these circulating H-T sperm-agglutinating antibodies, once they are formed, do not seem to interfere adversely with the germinal epithelium of the carrier.


The endothelium of failed corneal grafts. Corneal buttons of failed grafts were obtained at the time of regrafting and studied with the scanning electron microscope. The condititions studied varied from failed donor tissue to graft rejections and herpetic endothelial disease. The scanning electron microscope offers a tridimensional, panoramic high-power view of pathologic endothelium that can be correlated to the clinical picture and light or transmission electron microscopy. The endothelium of recent grafts may be damaged by organized blood clots; leukocytes were present over the graft endothelium, possibly as a normal reaction to surgery. Herpetic infection of the graft may cause death of endothelial cells and formation of granulomatous lesions, with damage to Descemet's membrane. A large detachment and rupture of Descemet's membrane was found in a case of wound dehiscense.


Immunologic  aspects of cicatricial pemphigoid. A 56-year-old woman presented with the typical findings of cicatricial pemphigoid: vesicular lesions of the mucous membranes and skin. Ocular involvement included symblepharon and dryness. A regimen of 80 mg of oral trimcinolone per day healed the lesions of the skin and mucous membrane. The patient received three injections of 0.5 ml of triamcinolone acetonide into the area of the symblepharon without apparent effect on the ocular disease. Direct immunofluorescent microscopy of the patient's conjunctiva revealed fluorescence of the basement membrane when incubated with fluorescein labeled anti-IgG. The finding of immunologlobulin fixed to be basement membrane in cicatricial pepmhigoid indicates some type of immune mechanism, and would seem to place this entity in the spectrum of the pemphigus-pemphigoid group.


Capillary density in developing and healing tuberculous lesions produced by BCG in rabbits. A quantitative study. Dermal tuberculous lesions were produced in rabbits by the intradermal injection of BCG. At various times after infection, anesthetized animals were perfused with a gelatin-colloidal carbon medium via the abdominal aorta. The capillary density of the nonnecrotic granulation tissue in the lesions was determined quantitatively by counting the capillaries under an ocular grid of a microscope. The capillary density in normal skin near the lesions was 3.8 plus or minus 0.5 in millimetersof capillary lengths per square millimeter in 250-mu tissue sections. The capillary density of the nonnecrotic tissue in BCGlesions averaged 6.1 plus or minus 0.6 mm/sq mm, an increase of 60%. The capillary density remained more or less constant as the BCG lesions grew and then regressed. The development of delayed hypersensitivity seemed to increase the capillary density, but this increase may have been a response to an extension of the necrosis at the time delayed hypersensitivity developed. Capillary densities in tuberculin reactions resembled those in BCG lesions. In the early stages, the increaseed capillary network of dermal BCG lesions was derived mainly from the subpapillary vascular plexus of the deep dermis supplied branches that surrounded the lower half of the caseous necrotic center and anastomosed with capillaries from the subpapillary plexus supplying the upper  half. When the necrotic center extended, nearby capillaries thrombosed and in turn became necrotic. Peripherally, new capillaries formed and anastomosed with existing capillaries. From these vessels, mononuclear phagocytes emigrated, destroyed the tubercle bacilli, and enabled the lesion to heal. In the BCG lesions at all stages of development and healing, the capillary network in the nonnecrotic areas seemed adequate to supply and nourish the defense cells controlling the infection.


An analysis of the Phagocytic potential of multinucleate foreign body giant cells. Multinucleate giant cells were collected by subcutaneous implantation of plastic films into mice. The attached cells were challanged in vitro with staphylococci, yeasts and sheep erythrocytes treated with either glutaraldehyde or isologous or heterologous antiserum. Cells containing more than seven nuclei rarely phagocytized yeasts or staphylococci, and the uptake and ingestion of sheep erythrocytes treated with heterologous antiserum was equally infrequent. Many sheep erythrocytes treated with isologous antiserum or glutaraldehyde attached to giant cells. When the adherent erythrocytes were related to the increased size of the multinucleate cell by dividing the number attaching by the number of nuclei in the giant cell, a progressive relative reduction was demonstrated as the nuclear content increased. It is suggested that these phenomena are due to the loss of surface receptors subsequent to fusion during the formation of multinucleate cells.


Natural selection on the dentition of an Arikara population. The tooth crowns of most mammals do not change in size except by attrition. Therefore, by comparing age groups within a population and finding differences in size or variation, the effects of such agencies as directional or stabilizing selection can be demonstrated. Mesiodistal and buccolingual measurements were taken on the maxillary and mandibular permanent dentition of a protohistoric Arikara Indian population from South Dakota. The tooth sizes of juveniles ages 6 through 15 were compared to those of adults ages 16 through 20. The two age groups were also compared by the cross-sectional areas of the posterior teeth. When a difference between age groups was demonstrated in either size or variation, a selection intensity was estimated using published graphs. Results from the comparisons show that adults, in general, have larger and less variable teeth than juveniles. The data suggest the possibility that both directional and stabilizing selection were operating on the Arikara dentition. Furthermore, the selection intensities suggest that selection was operating not on the individual teeth but on functional complexes such as the posterior teeth. It appears that in populations like the Arikara where crown attrition is severe, selection will be in the direction of large teeth.


Otosclerotic stapedial footplate fixation in an 18th century burial. Fixation of the footplate of the stapes by otosclerosis had been found in the left ear of a probable Spaniard who died about 200 years ago and was buried in the Tuscon Presidio. Previous studies on North and South Dakota American Indian crania of about the same time period did not demonstrate any stapedial fixation in 2,600 burials (4,064 temporal bones), 1,416 of which were over 16 years of age at death. The Tucson burial would indicate that this abnormality can be demonstrated in old interred skeletal material. Our other studies suggest that this disease process was not common in the prehistoric and historic Indian populations of the Dakotas.


The language capability of Neanderthal man. Considerable publicity has been given a recent article by Lieberman and Crelin ('71) which argues that Neanderthal man lacked the physical features necessary for speech. This present paper presents statistics on some modern men with normal speech who show physical features in common with Neanderthal man. The brain of Neanderthal man was as large as, or larger than, that of modern man. The brain reflected by endocranial cast of the La Chapelle-aux-Saints skull resembles that of modern man in an area important for speech and therapy suggests Neanderthal man had the neural development necessary for language.


Cribra orbitalia in the aborigines of Hawaii and Australia. Cranii of 53 Hawaiian aboriginal infants and children, and 45 from Austrailian aboriginal children were inspected. Cribra orbitalia was present in 22.8% of the former and 26.6% of the latter; osteoporotic pitting (symmetrical osteoporosis; porotic hyperostosis) was also present in the latter. The frequency compares favorably with that found in pre-Columbian North American Pueblo Indians, 24.7%. It is associated with a widespread skeletal involvement suggestive of an active bone marrow. The findings support the concept that cribra orbitalia is related to symmetrical osteoporosis and that it may be associated with a blood disorder.


Metrical analysis of Fontechevade II.

Metrical analysis of Fontechevade II. The Fontéchevade pi calotte is analyzed by principal coordinates on the basis of 12 measurements. It most closely resembles Classic Neandertals and the Steinheim cranium in shape, showing no special similarity to modern man.


Kinetics of unidirectional leucine transport into brain: effects of isoleucine, valine, and anoxia. The rate of unidirectional uptake, v, of L-[4,5-3H]leucine was studied in 17 isolated dog brains by means of an indicator-dilution technique using 22Na as the intravascular reference. The arterial leucine concentration was varied in increments by adding unlabeled leucine to the blood and v was determined after each change. The valine and isoleucine concentrations were varied independently to permit evaluation of their effect on leucine transport. A preliminary analysis indicated that both valine and isoleucine are competitive inhibitors. Therefore, all data were fitted to an equation that describes Michaelis-Menten kinetics in the presence of two competitive inhibitors. These calculations yielded an apparent Km for leucine transport of 1.58 mM plus or minus .28 SE, a V max of 0.323 mumol/g per min plus or minus .035 SE, and an apparent K i for inhibition of leucine transport of 1.76 mM plus or minus .34 SE for valine and 0.73 mM plus or minus .14 SE for isoleucine. In four isolated brains perfused with blood having a constant leucine level, indicator-dilution injections were made before, and at 1, 5, and 10 min after the start of perfusion with anoxic blood. These findings showed that, unlike glucose transport (Brain Res. 67: 307-316, 1974), the rate of lucine transport is unaffected by 10 min of anoxia.


Lipoate effect on carbohydrate and lipid metabolism and gastric H+ secretion. Acid secretion (QH+) and oxygen consumption (Qo2) by frog gastric mucosae in vitro were sharply stimulated by lipoate. A rapid decline followed stimulation, subsequently falling below control values. Addition of only glucose or lactate had no effect on Qo2 or QH+. Pyruvate caused slight significant stimulation of Qo2. Any one of these compounds added to lipoate-treated mucosae increased the stimulatory effect of lipoate and markedly slowed the rate of decline subsequent to maximum stimulation. Various fatty acids had a moderate-to-high stimulatory effect on Qo2 and QH+. Lipoate added prior to the addition of fatty acids decreased the stimulatory effect of buryrate (minus 56%), decanoate (minus 87%), and palmitate (minus 60%). Propionate became an inhibitor in the presence of lipoate. Lipoate increased (plus 100%) the amount of glycogen oxidized and decreased (minus 69%) the amount of triglycerides oxidized. Lipoate-treated mucosae did not respond to histamine. Addition of glucose restored responsiveness. The results indicate that beta-oxidation of fatty acids plays a major role in the acid secretory process and is centrally involved in cyclic AMP and histamine stimulation of QH+.


One-year follow-up of hyperactive boys treated with imipramine or methylphenidate. The authors report on a one-year follow-up of 76 hyperactive boys who had participated in a comparative study of methylphenidate, imipramine, and placebo. They found that there was a higher rate of discontinuance of imipramine than of methylphenidate in this sample and that both medications decreased the boys' rate of weight gain but had no significant effect on growth in height. This findings is of concern because larger doses than were used in this study are in use elsewhere. Teachers reported that the group of boys who had discontinued either medications showed continued hyperactivity and behavior problems in the classroom at one year. The two treatment groups showed continued improvement at one year in classroom and home behavior and did not suffer significantly from each other.


Cervico-mediastinal haematoma following sneezing. A case of spontaneous cervico-mediastinal haematoma is described. This is the ninth case recorded and is unusual in that the haematoma seems to have resulted from rupture of a para-oesophageal vein. The differential diagnosis and methods of investigation are discussed.


Post-operative convulsions due to penicillin applied to the spinal theca. Two cases are described and discussed in which penicillin powder applied to the theca at the end of laminectomy caused delayed convulsions.


Positive end-expiratory pressure during mechanical ventilation of the lungs.  A new vlave. A valve is described which will provide positive end-expiratory pressure during mechanical ventilation of the lungs. The authors will be pleased to supply further details of specification and production on request.


Chloroform or ether-a literary question. Thackeray's accurate knowledge of the pharmacology of chloroform and ether is considered and his source discussed.


Structural modifications of lutein cell gap junctions during pregnancy in the rat and the mouse. By use of lanthanum tracer and freeze-fracture procedures it was found that granulosa-lutein cells of the pregnant mouse and rat ovaries are connected by gap junctions and septate-like zones of contact. Lutein cell gap junctions enlarge and become partially internalized by the end of the first week of gestation. Expansion of the gap junction domain appears to be due initially to intercalation of particles along borders of small gap junctions devoid of smaller non-junctional particles. The number of gap junction lined processes appearing at the cell border increases concomitantly with hypertrophy of the lutein cell during the second week of pregnancy. Strands of particulate or grooved membrane emanate from the margin of larger gap junctions undergoing interiorization. Most large gap junctions are intimately associated with elements of the smooth endoplasmic reticulum. Spherical gap junctional profiles assume a deeper location in the lutein cell and may form concentric arrays by term while true surface gap junctions appear to fragment in the post-partum corpus luteum. The modifications observed are interpreted with respect to biogenesis of the gap junction and the hormonal control of lutein cell function.


SEM preparation and observations of the hen's oviduct. The surface patterns of the oviduct of Gallus domesticus were examined by scanning electron microscopy (SEM) using several preparatory techniques. Ciliated epithelial cells predominate throughout the oviduct with non-ciliated cells approaching an equal proportion in the magnum and isthmus. Short, randomly oriented mucosal folds which characterize the infundibulum increase in height and longitudinal orientation as they approach the magnum. Here and in the isthmus they broaden and become dimpled with glandular apertures. The folds narrow again in the uterus and vagina. Cryofracture followed by SEM demonstrate the deeper mucosal folds and submucosal tubular system.


Lung volumes and closing capacity with continuous positive airway pressure. Total lung capacity, vital capacity, residual volume, and functional residual capacity were determined by body plethysmography and the single-breath oxygen (SBO2) test was performed at 0, 5, and 11 cm H20 continuous positive airway pressure in healthy, awake, seated, spontaneously breathing subjects. Mean values for the absolute lung volume at which phase IV of the SBO2 test begins (closing capacity) did not change significantly with continous positive airway pressure at 5 or 11 cm H2O. Mean total lung capacity, functional residual capacity, and residual volume increased significantly, and the mean closing volume, the lung volume above residual volume at which phase IV begins, decreased significantly with 11 cm H20 continuous positive airway pressure; differences at 5 cm H20 were not significant. The slope of the alveolar nitrogen plateau (phase III) obtained during the SBO-2 test did not change with continuous positive airway pressure.


Blood components in the treatment of acute blood loss: use of freeze-preserved red cells, platelets, and plasma proteins. To avoid untoward reactions from blood transfusions and to make best use of the limited supply of blood, anesthesiologists and surgeons have many newly developed means at their disposal. Blood components should be separated from whole blood at the time of collection and prepared for either liquid or freeze-preservation. Citrate-phosphate-dextrose (CPD) blood should be separated into its components at room temperature within 4 hours of collection for greatest service from each collected unit. Red cell concentrates with hematocrits of 70 volumes percent can be prepared from the whole blood at the time of collection and frozen either shortly thereafter or after storage at 4 degrees C. for up to 3 weeks. Red-cell levels of 2, 3-diphosphoglycerate (2, 3-DPG) and adenosine triphosphate (ATP) can be increased by a rejuvenation process prior to freeze-preservation with either 40 percent W/V glycerol and storage at minus 80 degrees C. or with 20 percent W/V glycerol and storage at minus 150 degrees C. While hemorrhagic shock can best be managed with fresh whole blood, such blood is often not available; liquid- and freeze-preserved products serve as best substitutes. When previously-frozen washed red cells are used, crystalloid, colloid, coagulation factors, and platelets may also be required. Platelet concentrates stored at 4 degrees C. provide platelets that are hemostatically effective immediately upon infusion but have poor circulation. Platelet concentrates stored at 22 degrees C. provide platelets that have good circulation but upon transfusion have impaired hemostatic effectiveness. The coagulation factors and oncotic properties of plasma protein necessary for proper treatment of patients in hemorrhagic shock can be met by an adequate supply of fresh-frozen plasma and albumin. When liquid-stored red-cell concentrates or whole blood is given, filters must be used to remove the accumulated amorphous material, although the actual effects of the infused microaggregates are not yet known.


"Hyper-" end-expiratory pressure in the treatment of adult respiratory insufficiency: a case report. Postiive end-expiratory pressure (PEEP), in combination with mechanical ventilation, is efficacious in the treatment of some forms of adult respiratory insufficiency. PEEP in excess of 10 to 15 torr usually is not recommended; however, higher levels of PEEP may be required to reverse the pulmonary venous admixture. The efficacy of optimizing cardiopulmonary function with varying levels of PEEP is described.


Fibrinogen and albumin deficiencies associated with packed red blood cell transfusions. Fibrinogen content was determined for each of 50 units of citrate-dextrose-phosphate (CPD)-preserved whole blood, packed red blood cells reconstituted with 250 ml. of saline, and packed red cells reconstituted with 250 ml. of purified plasma protein fraction (PPF). The total protein and albumin were measured, by electrophoresis, on each of 10 units of the three varieties of blood. The fibrinogen content of the two types of reconstituted cells was significantly lower than that of whole blood. Although the total protein/albumin content of whole blood and PPF-reconstituted red cells was similar, saline-reconstituted cells were markedly deficient in both total protein and albumin. Low fibrinogen and platelet levels subsequent to transfusion with reconstituted packed red cells can lead to an erroneous diagnosis of disseminated intravascular coagulation. Administration of large quantities of saline-reconstituted packed cells could be an etiologic factor in postoperative interstitial pulmonary edema.


Effects of prednisone on gastric secretion and development of stomach lesions in swine. In pigs treated with various doses of prednisone, pathologic changes appeared in the glandular region of the stomach, mainly in the fundic area and rarely and less severely in the cardiac and pyloric mucosae. The characteristic lesion was hemorrhagic gastritis; this was associated with massive gastric hemorrhages in pigs given the larger doses of prednisone. Epithelial changes were found in the esophageal area, but there were no ulcers. In pigs with gastric fistulas, prednisone administered at dose levels of 10 mg/kg of body weight for 8 days produced a significant increase in the volume and the acidity of gastric secretion. Hexosamine output, but not concentration, increased significantly during the treatment period. Change in pepsin secretion was sit significant. Prednisone did not produce an immediate change in gastric secretion--there being a latent period lasting 1 to 2 days before marked changes were observed. After peak changes occurred (3rd or 4th day of treatment), acidity gradually decreased to pretreatment level toward the end of the treatment period. This indicates that although increased acidity may be involved in the development of lesions in the glandular region, other factors probably have a role also in the pathogenesis of steroid-induced stomach lesions in swine.


Activity of penicillin and novobiocin against bovine mastitis pathogens. Penicillin (P), novobiocin (N), and both (P-N) were evaluated in vitro against 143 clinical isolates of bovine mastitis, including, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis. Greater antistaphylococcal activities were demonstrated with N alone and P-N than with P alone. Streptococcus spp were more susceptible to P alone than to N alone, and the effectiveness of P-N corresponded with that of P alone. In vitro, P-N had a wider spectrum of antibacterial activity than did either P or N. Similar results were also obtained in vivo, using the mouse protection test. The P-N gave a greater protection rate in mice experimentally infected with S aureus and Str agalactiae than did P or N.


Diazepam and paraldehyde for treatment of severe delirium tremens. A controlled trial. Thirty-four patients with severe delirium tremens were allocated randomly to treatment with paraldehyde (10 ml rectally very 30 minutes) or diazepam (10 mg then 5 mg intravenously every 5 minutes) until they were calm but awake. Diazepam-treated patients became calm in one half the time needed to calm patients with paraldehyde. Half of the patients had delirium tremens in association with pneumonia, pancreatitis, or alcoholic hepatitis; these patients required twice as much paraldehyde or diazepam for initial calming as patients with delirium tremens alone. Maintenance of a calm state was accomplished easily with either diazepam, intramuscularly, or paraldehyde, rectally. Adverse reactions occurred in nine patients, all of whom had been treated with paraldehyde; these patients had greater degrees of fever, tachypnea, and tachycardia and required three times longer for initial calming than patients without adverse reactions. Diazepam given under this regimen is a safe and effective sedative for management of combative patients with severe delirium tremens.


Goodpasture's syndrome: normal renal diagnostic findings. A patient with Goodpasture's syndrome is described in whom pulmonary manifestations were dramatic, but in whom renal abnormalities were minor and only found on fluorescent and electron microscopy. His urine showed no proteinuria and no increase in cells in quantitative counts, and renal function was normal. It is suggested that there may be an indication for carrying out renal biopsies in patients with idiopathic pulmonary haemosiderosis and that this may lead to an early diagnosis of Goodpasture's syndrome.


Host failure in treatment of malaria with sulfalene and pyrimethamine. An individual infected with a multidrug-resistant strain of Plasmodium falciparum failed to respond to treatment with sulfalene and pyrimethamine. Subinoculation studies showed that parasite resistance to the drug combination was not present. Plasma levels of sulfalene and pyrimethamine in this individual were similar to those of three individuals, subinoculated from him, who were cured by the drug combination. Erythrocyte levels of sulfalene in this individual were similar to those in an individual, subinoculated from him, who was cured by the drug combination. After treatment with the drug combination, in vitro tests showed similar antimalarial activity in the serum of this individual in comparison with the serum of this individual in comparison with the serum of an individual subinoculated from him. The failure of this individual to respond to treatment with sulfalene and pyrimethamine is attributed to an undefined host factor (or factors) that appear(s) to be present in his erythrocytes.


Digitalis in acute myocardial infarction: help or hazard? The use of digitalis in acute myocardial infarction remains controversial because of disagreement regarding the electrophysiologic, hemodynamic, and metabolic risks and benefits. However, there is no convincing evidence of an increased incidence of arrhythmias following digitalis therapy. Direct measurement of left ventricle function shows improvement of impaired left ventricular performance. The increased metabolic cost of enhanced inotropy following digitalis is countered by reductions in left ventricular end-diastolic pressure and volume, especially when left ventricular failure is present. Extension of infarct size shown in the dog after administration of digitalis may occur only in that experimental model or only if cardiac failure is absent. We conclude that digitalis may be recommended following acute myocardial infarction if the usual indications, supraventricular tachyarrhythmias or cardiac failure, are present.


Guidelines for prevention of catheter-associated urinary tract infections. The most common site of nosocomial infection, involving more than 400 000 patients in this country annually, is the catheterized urinary tract. These infections cause significant morbidity, prolong hospital stay, increase hospitalization costs, and contribute to mortality caused by Gram-negative septicemia. Strict adherence to appropriate infection control procedures is necessary if the incidence and sequels of catheter-related infections are to be reduced. Based upon current understanding of the epidemiology of nosocomial urinary tract infections, guidelines for the prevention of catheter-associated urinary tract infections are presented.


Autoimmunity: present concepts.

Autoimmunity: present concepts. Autoimmune disease may be effected by humoral mechanisms (direct antibody effect or pathogenic antigen-antibody complexes), cell-mediated mechanisms, and possibly complement in the absence of antibody. In view of recent work, self-tolerance can no longer be explained by the clonal selection theory; cells that are capable of recognizing self-antigens and effecting an autoimmune response seem to be present in the normal individual but actively regulated by a process involving antigen-antibody complexes or suppressor T cells. The determination that a clinical state is an autoimmune disease cannot, therefore, be made only on the basis of the presence of autoantibody as this may be part of the normal and active state of immunoregulation of autoimmunity. Indeed, the absence of autoantibody may be the immunodeficient state that allows a full-blown autoimmune disease to occur. In view of this knowledge, the basis for the diagnosis of autoimmune disease and the rationale for treatment based on the objective of inhibiting all immune responses to the autoantigen require re-examination.


Detection and growth of enteropathogenic Escherichia coli in soft ripened cheese. The organism most frequently encountered during the 1971 outbreak of enteropathogenic Escherichia coli (EPEC) in soft ripened cheese was a strain that failed to ferment lactose broth within 48 h. Since existing methods for E. coli are dependent upon fermentation of this sugar, such strains can remain undetected, particularly when present in low numbers. Therefore a cultural testing procedure was developed to insure isolation of both lactose-positive and -negative strains. This method used GN broth, modified by substituting lactose and arabinose for glucose and D-mannitol, as an enrichment medium. MacConkey agar, used as a plating medium, was modified by substituting arabinose for half the lactose. The cultural procedure was used in conjunction with a fluorescent antibody method to screen cheese for the presence of presumptive enteropathogenic E. coli. Suspected isolates were subjected to further biochemical and serological testing and identified as members of specific serogroups. These methods were used for the analysis of over 2,000 wheels of cheese; over 10% of the samples tested were found to contain strains belonging to six different serogroups associated with diarrheal diseases. No attempt was made to confirm pathogenicity by in vivo tests. Enumeration of E. coli in cheese showed that numbers increased during storage. Cheese with less than 10 organisms/g initially increased to over 10-5 at room temperature and over 10-3 at 4 C within 10 days. With higher initial counts, levels up to 10-9 were found at 4 C. These studies showed that the high levels of E. coli encountered in these products cannot be used as a direct indicator of post-processing contamination.


Evaluation of methods for the isolation of Salmonella and Arizona organisms from pet turtles treated with antimicrobial agents. Turtles infected with and actively excreting Salmonella-Arizona organisms were treated with various concentrations of both Neo-Terramycin (N-Te) and Terramycin (Te) (Pfizer) for various periods of time and then tested for the presence of these pathogens by two methods, excretion and blending.  Turtles treated with 200 mug of Te per ml of container water for 9, 12, or 14 weeks, whereas when representative turtles from treatment groups were blended 72 h posttreatment these organisms were isolated from the whole turtle homogenate.  Salmonella and Arizona could be recovered from homogenate prepared from turtles treated for 7 and 14 days with 400, 800, or 1,000 mug of Te or N-Te per ml.  These findings suggest that the blending methods is more sensistive than the excretion method for the detection of Salmonella-Arizona in the treated turtle.


Proficiency test specimens for water bacteriology. A procedure for the preparation and distribution of simulated water specimens for coliform density testing is described.  Lyophilization of Escherichia coli in a cooked meat-glucose supporting substrate provides stable samples which may be distributed to participants in a proficiency testing program.  Logarithmic conversion of the data allows statistical evaluation of the results for inter- and intralaboratory variation.  Comparisons between the most probable number and membrane filter techniques indicate no significant difference in the accuracy of these techniques, although the membrane filter technique is more precise.


Length of incubation for enumerating nitrifying bacteria present in various environments. The effect of incubation time on most-probable-number estimates of autotrophic nitrifying bacteria was investigated by using waters, rooted aquatic plants, sediments, and slimes as inoculum sources. Maximum most probable numbers of the NH4+-oxidizing group were attained in 20 to 55 days (median, 25). Estimates of NO2-oxidizers were highest at termination (103 to 113) days.


Rapid gas chromatographic technique for presumptive detection of Clostridium botulinum in contaminated food. A simple gas-liquid chromatography end-product assay is reported for butyric and other short-chain fatty acids as presumptive indicators of Clostridium botulinum contamination in food.


Bacteriological survey of raw beef patties produced at establishments under federal inspection. At the time of manufacture, 76% of 74 sets of raw beef patties collected in 42 federally inspected establishments had aerobic plate counts of 1,000,000 or fewer/g; 84% contained 100 or fewer coliforms/g; 92% contained 100 or fewer Escherichia coli/g; and 85% contained 100 or fewer staphylococcus aureus/g (geometric means of 10 patties/set). Salmonellae were isolated from only three (0.4%) of 735 beef patties.


Effect of exposure to hyperoxic, hypobaric, and hyperbaric environments on concentrations of selected and aerobic and anaerobic fecal flora of mice. Alterations in selected aerobic and anaerobic fecal microflora of the mouse were determined during exposure to hyperoxic and normoxic hypo- and hyperbaric environments. Examination of fecal cultures obtained during exposure for 6 weeks to either 60 or 77% oxygen concentration 1 atmosphere absolute revealed little alteration in the aerobic or anaerobic flora. There appeared to be only a retardation in the reduction of the Klebsiella-Enterobacter flora which normally occurs after weaning. During exposure to hypobaric environments (100% O-2, 0.2 atmosphere absolute), significant alterations in concentrations of Escherichia coli, slow lactose fermenters, Klebsiella-Enterobacter, and enterococci were found in some instances. All alterations were toward increased concentrations. Variations in concentrations of different colony types of obligately anaerobic gram-positive (anGPR) and gram-negative (anGNR) rods cultured during the same experiments also occurred. One colony type of anGPR appeared to decrease while a second type increased in numbers. Concentrations of three colony types of anGNR were generally, but not always, increased. During hyperbaric exposure (2.8% O-2, 7.5 atmospheres absolute), increased concentrations of Klebsiella-Enterobacter, E. coli, slow lactose fermenters and enterococci were also noted. Changes in numbers of both colony types of anGPR, when occurring, were in the direction of lower numbers. Alteration in numbers of anGNR were in both directions but were more frequent in the direction of higher numbers. After return to normal air for 4 weeks of either hypo- or hyperbaric exposure, fecal concentrations of all organisms tended to revert toward control values with the exception of the anGPR which remained in lower concentrations after termination of the hyperbaric exposure. These observations indicate that, despite the great variation in the fecal flora among individual mice, it is possible to discover the effects induced by altered gaseous environments.


Bacterial survival in laundered fabrics. Bacterial survival was determined in linens (i) inoculated with Staphylococcus auerus (ii), taken from hospital isolation patients' beds, and (iii) used by students in their homes. Two different washers using temperatures of 38, 49, 54 and 60 C, respectively, for different times were empolyed along with a commercial tumbler dryer. Findings, after macerating the linens in Waring blender and enumerating on nonselective media, indicate that acceptable levels of survivors can be acheived in motel and hotel linens by an 8- to 10-min wash cycle at 54 C followed by adequate drying. However, it is recommended that a wash cycle with 60 C for 10 to 13 min be  employed for linens in health care factilities. The microbial significance of various laundering practices is discussed.


Bacterial flora of the hemolymph of the blue crab, Callinectes sapidus: most probable numbers. The hemolymph of 290 freshly collected blue crabs from Chincoteague Bay, Va., was sampled over a 15-month period from August 1968 through November 1969 and most probable numbers of bacteria were determined by tube dilution. The hemolymph of 18% of all crabs sampled was found to be sterile, with 16% sterility in summer and 23% in winter samples. Despite individual variations, male crabs as a group had a higher bacterial hemolymph burden than females, and among both sexes summer counts were higher than winter. The hemolymph of crabs with missing appendages had significantly higher counts than uninjuried crabs. The annual mean hemolymph most probable numbers per ml was 2,756 for males, 1,300 for females, and 1,876 for both sexes. The higher bacterial levels found in the hemolymph of male crabs may, in part, be explained by the fact that males, which predominated in the summer samples, had a higher incidence of injury and missing appendages than did females.


Bacterial flora of the hemolymph of the blue crab, Callinectes sapidus: numerical taxonomy. Bacteria isolated from the hemolymph of normal blue crabs were found to be predominantly Vibrio sp., Pseudomonas sp., Acinetobacter sp., Bacillus sp., Flavobacterium sp., and coliforms. Vibrio parahaemolyticus, a cosmopolitan facultative pathogen widely implicated in outbreaks of gastroenteritis related to the consumption of improperly processed seafood, was present in crabs collected between the months of May to November and was identified in up to 21% of the hemolymphs sampled.


Prophage induction of Escherichia coli (lambda) by N-nitrosamines. Carcinogenic N-nitrosamines were tested for their ability to induce lambda in a lysogenic strain of Escherichia coli K-12 (58-161 F+ Dimethylnitrosamine, di-n-propylnitrosamine, methyl-n-propylnitrosamine, and N-nitrosopiperidine were shown to be inducers of prophage.


Livedo vasculitis (the vasculitis of atrophie blanche). Immunohistopathologic study. Hyalinizing segmental vasculitis or livedo vasculitis (atrophie blanche) is a clinical entity with a distinctive immunohistopathologic morphology that can be distinguished from other forms of cutaneous vasculitis by histologic and direct immunofluorescent studies. Our studies showed that immunoglobulins and complement components (C-1g, C-3, and properdin) were localized in diseased vessel walls, suggesting an immune pathogenesis.


Antinuclear antibodies. II. The prognostic significance of nuclear immunofluorescent patterns in lupus erythematosus. Nuclear immunofluorescent patterns in lupus erythematosus (LE) were correlated with the clinical picture. Patterns were classified into three groups of prognostic significance: (1) Poor-Shrunken peripheral fluorescence was found in patients with severe systemic lupus erythematosus (SLE), 60% demonstrating renal involvement. (2) Intermediate-This included the peripheral and therapy patterns, associated with 33% renal involvement in patients with SLE. (3) Good-Large and small speckle-like threads were seen in cases of very benign LE.


Human melanoma FUHS-1. Description of a new cell line. Presented is a new established cell line, which was cultivated from a melanoma originating from a melanosis praeblastomatosa circumscripta of the leg. Recently subculture No. 105 was reached after a period of 31 month. Medium of primary culture was TC Medium Eagle spinner modified, followed from subculture No. 1 by MEM/Hanks/0.35 g/l NaHCO3. From subculture No. 6 NCTC 135 was applied simultaneously-each respective supplements. The cells grow as monolayer; absolute plating efficiency of the established cell line lies by 800/0; epithelial-like and dendritic melanin-producing cells are prominent. The number of chromosomes is considerably fluctuating; the karyotype of the established cell line lies within the range of subtriploidy. Sizewise a marker in submedian position of centromer is to be compared with one of group.


Macrophage migration inhibition test in untreated syphilis. Two modifications of macrophage migration inhibition test, one of George and Vaughan and the other one of Svejcar, were performed on a total of 78 cases of untreated syphilis at various stages. As specific antigens were used: Treponema Pallidum ultrasonate and cardiolipin. Inhibition of migration was observed in 87 percent patients with primary syphillis and in all patients with late and late congenital syphilis. After improving and standardisation of the technique of Treponema Pallidum antigen-the migration inhibition test may be recommended as a specific in vitro-test for detection of cell mediated immunity in syphilis.


Chrysanthemum allergy. Pt. II: Experimental studies on the causative agents. Experimental studies on allergic contact dermatitis due to Chrysanthemum indicum 1. (Chrysanthemum x-morifolium) (chrysanthemum of the florists) have indicated the following results: 1. In 5 patients with allergic contact dermatitis from chrysanthemum oil of turpentine and its sensitizing compounds gave no patch test responses. No relationship between contact allergy due to chrysanthemus and to turpentine oil could be determined. 2. The pyrethrins, constituents with insecticidal activity, derived from certain Chrysanthemum species and often suspected as the causative agents, play no role in chrysanthemum allergy. 3. Tests on sensitized guinea pigs (pirl bright white strain) with flowers of chrysanthemum as well as with the two sesquiterpene lactones parthenolide and alantolactone, derived from different Composite species, gave positive patch test reactions. The results showed that parthenolide produced stronger reactions than alantolactone. 4. By thin layer chromatography neither parthenolide, nor alantolactone or pyrethrosin could be detected in extracts of chrysanthemum flowers of the florists. But the investigations indicated that several other terpenic compounds are present, which gave positive color reactions to certain lactone reagents. Five of them showed strong positive patch test reactions in our patients as well as in sensitized guinea pigs. Further studies are required to identify these compounds.


Humoral immune response of mice with long-term exposure to cigarette smoke. Mice were exposed to fresh cigarette smoke for 1,17, and 38 weeks. After each period the primary and secondary humoral immune response to intraperitoneal inoculation of sheep erythrocytes was studied. Direct and indirect plaque forming cell responses in the spleen and in a pool of cervical and mediastinal lymph nodes, as well as serum hemolytic and hemagglutinating antibodies were examined. Primary plaque forming cell and antibody response to sheep erythrocytes was first enhanced and then depressed by continual cigarette smoke exposure, while the secondary response was unaffected. Serum antibody response to polyvinylpyrrolidone, an immunogen independent of thymus-derived lymphocytes, was not impaired by 41 weeks of cigarette smoke exposure.


Cardiac surgery in patients undergoing renal dialysis or transplantation. The coexistence of organic cardiac disease and chronic renal failure presents a therapeutic dilemma. Cardiac operations have been performed on patients who were undergoing hemodialysis or who had had successful renal transplantation. There are several aspects of management of these patients which differ from those of the routine cardiac surgical patient. Guidelines for management are outlined and discussed.


The benefits of corticosteroids in endotoxic shock. The experiments reported here were undertaken to study the effects of pharmacological doses of corticosteroids administered alone or in conjunction with prolonged (12-hour) assisted circulation in 22 dogs subjected to LD50-60 Escherichia coli endotoxin. The most striking findings were lengthened survival time, higher cardiac output, decreased fluid requirement, and minimal evidence of pulmonary congestion or injury in the animals treated with steroids only. Unexplained mesenteric infarction prematurely terminated the experiments in animals undergoing assisted circulation. The benefits of corticosteroids in experimentally induced endotoxic shock are clearly demonstrated in these experiments. Further studies are needed to clarify the supportive role of assisted circulation in endotoxic shock and to determine any possible advantage of hypothermia over normothermia during its course.


Aromatic metabolism in the fungi. Growth of Rhodotorula mucilaginosa in p-hydroxybenzoate-limited chemostats and the effects of growth rate on the synthesis of enzymes of the 3-oxoadipate pathway. Rhodotorula mucilaginosa was grown in p-hydroxybenzoate-limited chemostats over the dilution rate (D) range 0.01 to 0.17 h-1 and growth was adequately described by the Monod theory when maintenance energy requirements were considered. The p-hydroxy-benzoate affinity constant, K8, had the relatively high value of 270 mg/I. The yield from p-hydroxbenzoate varied with dilution rate but was constant above D equal 0.07 h-1 at 0.56 g yeast/g substrate utilised. The maintenance coefficeint for growth on the aromatic substrate was 20 mg/g yeast/hr. Culture viability decreased linearly as the dilution rate was reduced. 4-Hydroxybenzoate 3-mono-oxygenase, protocatechuate 3,4-dioxygenases, 3-carboxymuconate cyclase and 3-carboxymuconolactone hydrolase activities were dilution rate-dependent, results which accord with the substrate in inducibility of these enzymes. Under carbon-limited growth conditions the addition of glucose, a catabolite repressor of these enzymes, to the aromatic medium stimulated their synthesis. Data were also obtained which indicated that whereas the synthesis of the cyclase and the hydrolase was coordinately controlled, that of the first two enzymes of the 3-oxodipate pathway was under independent control.


Accuracy of the family history method in affective illness. Comparison with direct interviews in family studies. We interviewed available spouses and first-degree relatives of 140 bipolar and unipolar probands for current and past psychopathology, and assessed interrater reliability. Diagnoses based on direct interviews of relatives were compared with those based on reports of the probands and of all other interviewed family members. Probands underestimated the prevalence of affective illness and other psychiatric disorders in their relatives, and overestimated the age of onset of illness in their ill relatives. Probands reported more accurately about illness in their spouses and parents than in their siblings and children, but accuracy reached acceptable levels for spouses only. Diagnoses on relatives derived by combining reports of all other interviewed family members, including the proband, were slightly more accurate than those based on the proband's reports alone. Good accuracy was obtained only for reports about spouses.


Heterotopic bone formation and alkaline phosphatase. Heterotopic bone formation is osteogenesis that occurs in a part of the body that does not normally form bone. Alkaline phosphatase (orthophosphoric monoester phosphorylase) has a wide substrate specificity for catalyzing the hydrolysis of phosphate esters at an alkaline pH. The various isoenzymes are found to originate in the liver, bones, intestinal mucosa, and placenta. Numerous disorders are associated with elevations of the individual isoenzymes. The physiologic function of alkaline phosphatase is as yet unknown. The bone isoenzyme (heat labile) is found elevated in cases of heterotopic bone formation and fracture repair. Osteoblasts (bone forming cells) are unusually rich in alkaline phosphatase but the relationship of this enzyme to the process of calcification and bone formation is not clear. Alkaline phosphatase appears to be involved with the elaboration of matrix rather than with the process of mineralization in osteogenesis.


A new antireflux procedure at the esophagogastric junction: experimental and clinical evaluation. In posterior fundoplasty, a derivative of Nissen fundoplication, the fundus is passed behind the abdominal esophagus and fixed to the anterior aspect of the stomach, thus modifying the angle of entrance of the esophagus, creating a posterior gastric reservoir relatively inaccessible to the cardia. In dogs, it was a successful as a Nissen fundoplication in reversing esophagitis caused by reflux and daily administration of histamine. In another group of dogs, a cylindrical resection of the last 4 or 6 cm of the esophagus and a posterior fundoplasty were performed simultaneously. In spite of the daily administration of 30 mg of histamine, none of the dogs developed esophagitis.  Posterior fundoplasty has proved effective in 14 patients with hiatal hernia observed for three years. In five patients undergoing a Heller myotomy for achalasia, it was used successfully to prevent postoperative regurgitation.


Acinic cell carcinoma: collective review, including bilateral cases. Bilateral acinic cell carcinoma of the parotid gland is rare. On the basis of one case and a review of 486 parotid acinic carcinomas, unilateral as well as bilateral, total parotidectomy seems to be the treatment of choice for this low-grade malignant neoplasm that recurs commonly following inadequate excision. If involved by tumor, the facial nerve must be taken. Metastases are more likely to lung and bone than to regional nodes. For this reason, radical neck dissection is not routinely indicated.


Antilymphocyte globulin in renal transplantation: Nephrotic syndrome and infection as possible complications. During a two-year period, nine patients received equine antilymphocyte globulin (ALG) as part of their immunosuppressive therapy following cadaver renal transplantation. Of these, three patients (33%) developed nephrotic syndrome. A fourth patient had proteinuria greater than 3 gm/24 hr not attributable to recurrent disease or rejection. Of 32 other patients with cadaver renal allografts done during the same time, none had nephrotic syndrome and two (6%) had proteinuria greater than 3 gm/24 hr, both with biopsy evidence of rejection. This suggests an association between ALG therapy and risk for developing nephrotic syndrome, but the mechanism is not clear and deserves further investigation. During the first three months following transplantation, the nine patients treated with ALG had 20 infections, which seems to be more per patient than in our other transplant recipients.


Electrosurgical excision of full-thickness burns. Massive intraoperative blood loss and poor graft take have been the major problems associated with early excision and immediate grafting of full-thickness burns. By employing electrosurgery, excessive blood loss was virtually eliminated in a series of major burn excisions. Immediate graft take was excellent on electrosurgical wounds after primary burn excisions and in late reconstructive procedures. Simplicity, improved hemostasia, good graft take, and the absence of special anesthetic requirements make this method particularly applicable to the management of patients with burn injury.


Intestinal necrosis and perforation after renal transplantation. An acute intestinal disorder occurred in seven patients soon after renal transplantation. The disorder was characterized initially by massive abdominal distention with progression to multiple bowel perforations in three patients and extensive bowel necrosis in two. Surgical manipulation did not appear to be the cause since the transplants were performed retroperitoneally in the iliac fossa. Ischemia also was not a factor since the mesenteric vessels were patent with no evidence of thrombus formation or vasculitis. The cause appeared to be related to immunosuppressive agents, in particular, high doses of corticosteroids. The disease process appeared to be reversible if immunosuppression was discontinued early, as occurred in two patients.


Transsacral repair of rectal prolapse.

Transsacral repair of rectal prolapse. Whatever the basic pathogenesis of rectal prolapse, the structural alterations invariably include a defect in the pelvic floor fascia, a deep pouch anterior to the rectum, a patulous anus, loose fixation of the rectum to the pelvic structures, and a long sigmoid colon. A variety of corrective procedures have been described for rectal prolapse, but all suffer from various disadvantages. The transsacral approach to the rectum has several advantages: (1) it is simple and safe even for the elderly, (2) it avoids an abdominal incision with its concomitant postoperative complications, (3) it will allow complete correction of all abnormalities, and (4) it has been performed with a low recurrence rate.


An intraluminal technique for arterial occlusion during renal transplantation. A simple technique was developed for occlusion of atherosclerotic vessels in patients undergoing renal transplantation. Fogarty catheters were placed within the iliac arteries via a stab wound and the balloons inflated to provide safe and gentle occlusion of atherosclerotic vessels during the transplant operation.


Optic nerve anomalies in basal encephalocele. Basal encephalocele should be suspected in child when hypertelorism, defect of the bridge of the nose, and other midfacial anomalies are present. Optic nerve anomalies may occur with basal encephalocele. Those previously described are pale discs, coloboma of the optic nerve head, and pit of the optic disc. Two additional disc anomalies that are associated with basal encephalocele are megalopapilla and optic nerve dysplasia. A mass in the nose or nasopharynx under these circumstances is most likely an encephalocele and biopsy is contraindicated.


Posterior uveitis and primary cerebral reticulum cell sarcoma. Primary cerebral reticulum cell sarcoma appeared as unexplained posterior uveitis. A retrospective review of 19 cases of cerebral reticulum cell sarcoma seen at Massachusetts General Hospital has not confirmed the previously described high incidence of ocular involvement in patients with this tumor. The presence of posterior uveitis remote from a definite cerebral mass should suggest the possible diagnosis of primary cerebral reticulum cell sarcoma.


Reconstruction of the central one half of an eyelid. The technique recommended in this communication concerns the use of a semicircular graft from the lateral canthal area to close a lid margin defect comprising one half of the lid. When the defect is closed, the semicircular flap will straighten and become the lateral portion of the lid.


Eye manifestations in acrodermatitis enteropathica. A case of acrodermatitis enteropathica is presented that is the first to manifest punctal stenosis, the fourth to show corneal changes, and the sixth to survive into adulthood.


Behavioral studies using genetically defined mice. A bibliography. References in this bibliography have been selected from the Subject-Strain Bibliography of Inbred Strains of Mice, maintained at The Jackson Laboratory, which attempts to include all published papers dealing with specific inbred strains of mice, named genes in mice, or named transplantable tumors. We have selected all references which appear to be of behavioral interest, including reports of the effects of neurological mutations, but have omitted genetic studies conducted with these mutants. Studies using "white", "Swiss", or undesignated mice are not included. This bibliography covers literature published from 1922 through late 1973. The authors would like to be informed of omissions, and to receive reprints of omitted papers. The bibliography is divided into three sections. The first section includes all references in which a behavioral measure appears to be the variable of primary interest. This section is divided into 16 behavioral categories: activity, aggression, audiogenic seizures, communication, emotionality, feeding, learning, maternal, memory, psychomotor, regulation, reproduction, biorhythms, sensation, social, and miscellaneous. References are assigned to a category on the basis of their apparent emphasis. The second section includes all references in which the effects of a treatment on behavior appear to be the variables of primary interest. This section is divided into nine treatment categories: age, alcohol, central nervous system, mutations, neonatal and teratogenic, population size, pharmacological agents, genetic selection, and miscellaneous. Where multiple treatments were used, references are assigned to the category of the most important treatment. The third section contains reviews and theoretical references. Each item in the bibliography is assigned to a category in one of the three sections and given a reference number. At the end of each category is a list of the reference numbers from other categories which contain information pertinent to that category. References are arranged alphabetically within each category.


The action of bacterial cytidine deaminase on 5,6-dihydrocytidine. Cytidine deaminase from Escherichia coli was found to catalyze the hydrolytic deamination of 5,6-dihydrocytidine, at a rate slightly lower than its rate of action on the normal substrate. The results suggest that nucleophilic addition by the enzyme at the 5,6 position of the substrate is not an essential part of catalysis, unless the active site is so flexible that deamination can occur with addition in one case (cytidine) and without addition in another case (5,6-dihydrocytidine). 3,4,5,6-Tetrahydrouridine bears a close structural resemblance to a hypothetical "tetrahedral" intermediate formed by direct water addition to 5,6-dihydrocytidine. The hydrolytic activity of the enzyme toward 5,6-dihydrocytidine and its potent inhibition by 3,4,5,6-tetrahydrouridine are presumably related by the ability of the active site to stabilize structures of this kind by tight binding. Cytidine deaminase shows no detectable activity as a catalyst for the dehydration of 6-hydroxy-5,6-dihydrouridine.


DNA-protein interactions of the rat liver non-histone chromosomal protein. Native rat liver NHC protein-DNA interactions have been investigated by use of a nitrocellulose filter assay sensitive in detection of protein-DNA complexes. Optimal conditions for DNA-protein interactions occurs at low ionic strength conditions (110 mM phosphate buffer). A fraction of NHC proteins was enriched 25-fold by their affinity for rat DNA immobilized on cellulose columns under these conditions. At higher ionic strength (260 mM-0.04M phosphate buffer and 0.15 M sodium chloride), this fraction binds approximately sevenfold less to rat DNA but with a substantial increase in stability of the complexes. Equilibrium competition experiments indicate that at the higher ionic strength there is a considerable DNA sequence specificity of the rat DNA binding NHC protein. Since rat DNA contains three components as defined by their reassociation kinetics: single copy DNA (C0t1/2pure = 1.6 times 103); middle repetitive DNA (C0t1?1PURE = 1.1); and highly repetitive (C0t1/2pure smaller than 0.02). The two former were isolated and employed in the DNA binding assays. At the high ionic strength criterion, the rat DNA binding NHC proteins showed a substantial preference for a subset of middle repetitive DNA sequences. This suggests a preferential interaction between a class of NHC proteins and a class of middle repetitive DNA sequences.


Stepwise enzymatic oligoribonucleotide synthesis including modified nucleotides. A method has been developed for the routine synthesis of 2'(3')-o-monoacyl ribonucleoside 5'-diphosphates for stepwise synthesis of oligoribonucleotides with Escherichia coli polynucleotide phosphorylase. The use of triethyl orthoisovalerate allows the facile preparation of 2'(3')-o-isovaleryl-UDP, -CDP, -ADP, -GDP, -IDP, -EPLISON-APD, eplison-CDP, and N6-isopentenyl-ADP. The synthesis of N6-isopentenyl-ADP from ADP by N1-alkylation and the Dimroth rearrangement to N6 is reported. The effects of several factors including the nature of the divalent cation, pH, SALT CONCENTRATION, AND TIME ON THE EFFICIENCY OF THE POLYNUCLEOTIDE PHPSPHORYLASE CATALYZED SINGLE ADDITIONS OF THE 2'(3')-O-ISOVALERYL RIBONUCLEOSIDE 5'-DIPHOSPHATES TO AN OLIGORIBONUCLEOTIDE PRIMER ARE REPORTED. The syntheses of many tetranucleoside triphosphates and two pentanucleoside tetraphosphates in yields of 20-75 per cent are reported. The 2'(3')-o-isovaleryl derivatives of IDP, eplison-ADP, eplison-CDP, and N6-isopentenyl-ADP were all accepted by polynucleotide phosphorylase as substrates for the monoaddition reaction. The extension of the method to include the syntheses of oligoribonucleotides containing modified nucleosides offers a means of studying the role s of these modification by the use of relatively simple model compounds.


A permeable cell system for studying DNA replication in synchronized HeLa cells. Treatment of HeLa cells with a hypotonic buffer solution makes them permeable to nucleotides. Cells which are in S-phase at the time of treatment continue to synthesize DNA when supplied with the four deoxyriboside triphosphates, ATP, Mg2+, and the proper ionic environment. DNA replication extends from sites which were active in the cells prior to treatment. The product is confined to the nucleus and is sensitive to deoxyribonuclease. Under optimum conditions, up to 5% of the HeLa genome can be replicated from exogenous nucleotides. In synchronized cultures the level of DNA replicase activity, as measured in permeable cells at different points in the cell cycle, correlates with the rate of [14C] thymidine incorporation measured in the living, untreated cells.


Polypeptides of a glycoprotein antigen (SF) present in serum and surface of normal but not of transformed chicken fibroblasts. A protein immunologically identical to a glycoprotein antigen from fibroblast plasma membrane (SF antigen complex) is present in chicken serum. This antigen disappears from cells transformed with tumor viruses. The antigen solubilized from fibroblasts using urea and detergents, and the serum component both gave a molecular weight of about 2-10(5) as estimated by gel filtration on Sepharose 4B. Ultracentrifugation in 5-20% sucrose gradients gave a value of 7-8 S for the antigen from both sources. Isolation of antigen from serum and fibroblasts extracts was achieved using immunochemical techniques. Analysis of the polypeptide chains of the cellular antigen by electrophoresis in the presence of sodium dodecylsulphate, disclosed three major components at molecular weights 210 000 and 145 000 and 45 000. The two high molecular weight bands were identified in electrophoretograms of whole cell extracts by absorption to and subsequent elution from immunoadsorbents. The 45 000-molecular weight components comigrated with purified actin in electrophoresis. After short pulses of (35S) methionine the high molecular weight polypeptides, the 145 000-molecular weight component in particular, were among the most prominent radioactive bands from whole cell extracts indicating that SF antigen has a rapid turn-over. The antigen isolated from serum showed a polypeptide composition similar to the one from cells except the 45 000-molecular weight component was not detectable.


Azathioprine in rheumatoid arthritis: a long-term follow-up study. In 1973 we reported the beneficial effects of azathioprine in a double blind, cross-over study in 17 patients with classic rheumatoid arthritis. During subsequent follow-up over a mean period of 40 months, 4 patients had discontinued therapy because of poor therapeutic response and 1 because of nausea. Eleven of the 12 patients still taking azathioprine had maintained their initial beneficial response or showed further improvement. Adverse side effects during the follow-up period were minor. They included nausea in 1 patient and leukopenia with thrombocytopenia in another. An increased incidence of chromosomal abnormalities was detected in those patients still receiving azathioprine.


Polynucleotides. XXVIII. Stimulation of the binding of aminoacyl-tRNA to ribosomes by tri- and polynucleotide analogs. Messenger activity of synthetic tri- and polynucleotide analogs was studied by binding of 14C-labeled aminoacyl-tRNAs to ribosomes in the presence of the analogs. Synthetic messengers used were: poly(A) analogs in which adenosine was replaced by tubercidine (I), 3-deazaadenosine (II), 1-deazaadenosine (III) and 2-methyladenosine (IV); copolymers of adenosine and aristeromycin (V); cyclic triadenylate (VI); the heptanucleotide of 6,2'-O-cyclouridine (VII); the pentanucleotide of 8,2'-S-cycloadenosine (VIIIa); A-U-G analogs in which adenosine was replaced by 8,2'-O- and S-cycloadenosine (VIII), 8,5'-O- and S-cycloadenosine (IX); 8-oxyadenosine (x); 8-bromoadenosine (XI) and formycine (XII). Among these oligo- and polynucleotides, analogs which contained nucleotides of anti conformation having appropriate bases for Watson-Crick type hydrogen bonding stimulated the binding of corresponding tRNAs to ribosomes.


Steffimycin B, a DNA binding agent.

Steffimycin B, a DNA binding agent. The antibiotic steffimycin B binds to double-stranded DNA as evidenced by difference spectroscopy and an increase of the thermal stability of DNA in the presence of the antibiotic. Salmon sperm DNA-steffimycin B complexes show a drastic decrease in template activity for Escherichia coli DNA polymerase I but not for DNA-idrected RNA polymerase. The differences in template properties of poly[d(A-T)] and poly (dG) - poly(dC)-antibiotic complexes,respectively, for DNA polymerase I and RNA polymerase suggest that the antibiotic interacts primarily with adenine or thymine bases or both in double-stranded DNA.


Regulation of RNA synthesis in Escherichia coli. III. Degradation of guanosine 5'-diphosphate 3'-diphosphate in cold-shocked cells. Cold-shocked cells of Escherichia coli can degrade intracellularly accumulated guanosine 5'-diphosphate 3'-diphosphate (ppGpp). The rate of ppGpp degradation is governed, as in whole cells, by the spoT gene; a rapid breakdown reaction is associated with the presence of the spoT+ allele and at least a five-fold slower decay occurs in spoT-minus mutants. The two degradation reactions in shocked cells display the following similarities: (i) the rates of degradation are equivalent to whole cell estimates, (ii) both require a full complement of activated amino acids, (iii) both are dependent upon supplements in the reaction mixture which stimulate the availability of energy-rich compounds and (iv) neither is inhibited by concentrations of ribosomal antibiotics which severely restrict protein synthesis. Apart from characteristic rate  differences, decay of ppGpp in shocked cells derived from spoT-minus strains is discerned from spoT+ mediated decay in shocked cells by sensitivity to high concentrations of tetracycline and by manganese ion dependence.


Selection of patients for bone marrow transplantation in severe aplastic anemia. Despite androgens and intensive supportive care, satisfactory survival in severe aplastic anemia remains at 20% or less. Histocompatible bone marrow transplantation can restore normal hematopoiesis in approximately 40% of similarly severe individuals. Delay of transplantation for 3 wk after diagnosis allows time for proper evaluation and for many spontaneous recoveries. Further delay increases risks of fatal complications and decreases chances for successful transplantation while the incidence of spontaneous remission declines. When available, early histocompatible bone marrow transplantation may be the treatment of choice for severe aplastic anemia.


Culture of autologous bone marrow in diffusion chambers: effect of host irradiation. Autologous bone marrow (BM) cells were cultured in diffusion chambers (DC) implanted into whole-body irradiated, non-irradiated, or sham-irradiated goats. Proliferation was apparent in DC implanted in both irradiated and nonirradiated goats. However, cells in DC cultured in irradiated hosts increased in number beginning earlier, proceeded at a faster rate, and reached higher numbers than in DC in nonirradiated hosts. Growth enhancement could not have occurred as a result of radiation-induced immunosuppression in autologous hosts. The nonirradiated "target cells" in the DC therefore constituted an indicator system for stimulatory or inhibitory substances in the host. The simultaneous increase in the number of granulocytes in peripheral blood and in DC of irradiated hosts was paralleled by an initial rise in serum colony-stimulating factor (CSF). A second, prolonged period of severe granulocytopenia following irradiation of the host correlated with high levels of serum CSF. Increased numbers of megakaryocytes were seen in DC as thrombocytopenia developed in the irradiated host. DC erythropoiesis disappeared rapidly in nonirradiated goats; however, in DC of irradiated goats, the number of erythrocytic precursors increased exponentially during ablation of host erythroid marrow. Anemia did not develop in the host during the culture period. Proliferation of mononuclear cells in DC was markedly stimulated by irradiation of the host. Proliferation of macrophages appeared independent of host treatment. These observations provide strong evidence for diffusion of specific and/or nonspecific humoral hematopoietic stimulators from the host into the DC. This stimulation appears to be elicited and/or intensified by irradiation of the host.