Immunobiology of head and neck cancer: basic concepts.

Patients with squamous cancer of the head and neck have unique perturbations of the immune system. These patients have marked depression of cellular immunity even in the early stages of disease. The known facts about the immunobiology and the immunodeficiencies that commonly occur are discussed. Also reviewed are the relationships of the immunologic deficits to stage of disease, to prognosis, and to the suspected etiologic factors of smoking, alcoholism, and malnutrition. Highlights are given of current immunotherapeutic trials. To date, most of the successful efforts in both immunologic research and immunotherapy of head and neck cancer have dealt with general, rather than specific, immune reactions. Further development in the area of specific immune responses may permit more meaningful measurements of tumor-specific reactions, thus yielding useful tools for immunodiagnosis as well as providing more effective and precise methods of immunotherapy.


Orbital diagnosis.

Accurate localization and diagnosis of orbital lesions is necessary to select the appropriate treatment. A systematic method of evaluating a patient with orbital disease is presented. The most common orbital lesions among children and adults are listed, and abnormalities that mimic exophthalmos are described. The tests that are most useful in evaluating orbital disease are described in detail. Several case reports illustrate the value of ultrasonography and three-dimensional computed tomography in the localization and diagnosis of space-occupying lesions of the orbit. Special tissue-processing techniques can be performed on some tumors to aid in establishing a diagnosis and in planning treatment.


[Biochemistry of the developmental cycle of Triatoma infestans (Vinchuca). 2. Biosynthesis of fatty acids].

The biosynthesis of fatty acids in Triatoma infestans was studied in all the nymph stages and adult insects by feeding experiments with 1-14C acetate. In other experiments labeled acetate, 1-14C palmitate or 1-14C stearate were injected in the abdomen. In another group of experiments the abdominal non intestinal tissue of adult insects was homogenized and the supernatants of 10 000 X g and 105 000 X g were incubated with 1-14C acetyl-CoA. The incorporation of the labeling in the fatty acids was studiel by gas liquid radiochromatography. It was proved that the supernatant of 105 000 X g possessed a fatty acid synthetase able to synthesize de novo the saturated fatty acids of 10 to 18 carbons with a maximal yield in the myristic and palmitic acids. The fatty acid synthetase was active from Nymph L stage. The supernatant of 10 000 X g was able to synthesize fatty acids tentatively identified as eicosenoic and eicosadienoic acids. The T. infestans has also an enzyme that elongated palmitate to stearate. In the fraction precipitable between 10 000 X g and 105 000 X g was apparently present a desaturating enzyme that converts saturated to monoethylenic acids. The enzyme was apparently only active from Nymph II stage.


Techniques for the lesion of the vomeronasal organ of the rat.

Two technical procedures were developed in order to suppress the vomeronasal organ (V.O.) of the rat. The organ was approached through the nasal or oral cavity and electrolytic or thermic lesions were respectively performed. For electrolytic lesions the projection of the organ on the nasal bone was first determined. After opening the nasal bones, the Vs. Os. were lesioned by way of an electric current applied through an electrode introduced between the mucosa and nasal septum. For thermic lesions, the projection of the V.O. on the palate's surface was determined. After opening this area and the underlying part of the maxillary bone, both Vs. Os. were cauterized.


Culture characteristics of human granulocyte-macrophage progenitor cells and their regulation in the context of bone marrow transplantation.

Aspects of the granulocyte-macrophage progenitor cell (CFU-GM) assay not previously used in the content of bone marrow transplantation were discussed in terms of the additional information which could be derived. This focused on the characteristics and responsiveness of the progenitor cells to regulatory activities and other agents. In addition, Ia-like antigenic determinants were demonstrated on human "pre-CFU-GM", an assay which can be used to assess what appears to be a more immature progenitor cell.


Total body irradiation and autologous marrow transplantation as consolidation therapy for spontaneous canine lymphoma in remission.

Seventeen dogs with spontaneous generalized lymphoma in complete clinical remission induced with combination chemotherapy were given 1100 R total body irradiation (TBI) and autologous marrow grafts as consolidation therapy. Marrow was obtained immediately before TBI, stored at 4 degrees C, and infused immediately after TBI. Two dogs died because of failure to regain adequate marrow function, 8 died after developing recurrent lymphoma and 2 died of miscellaneous causes. Five dogs are alive in unmaintained complete clinical remission 200-663 days after initiation of chemotherapy. In a comparable group of 8 dogs in complete remission after combination chemotherapy but not given consolidation therapy, none remained in remission beyond 113 days. These results indicate that high dose TBI in conjunction with autologous marrow grafting results in prolonged remission duration in dogs with spontaneous lymphoma.


Autotransplantation after in vitro immunotherapy of lymphoblastic leukemia.

We have used in vitro immunotherapy before autologous bone marrow transplantation for three patients with acute lymphoblastic leukemia (ALL). Bone marrow was removed during remission, and mononuclear cells were separated by density-step centrifugation on Ficoll-Hypaque. The cells from each patient were treated with a heteroantiserum and complement to eliminate leukemic cells and were cryopreserved. Following chemotherapy and total body irradiation, the treated marrows were thawed and infused. All the patients showed positive evidence of returning marrow function before death. One patient who survived 4 months showed no evidence of leukemia at post mortem, and marrow sections demonstrated active hematopoiesis of all cell lines.


Autologous bone marrow transplantation in relapsed adult acute leukemia.

From March, 1976 to February, 1979, 28 cases of adult acute leukemia of which 24 were evaluable were treated in irreversible relapse with high dose chemotherapy (piperazinedione) and supra-lethal total body irradiation (TBI) in conjunction with autologous bone marrow transplantation (ABMT). The marrow cells grafted were collected and stored in liquid nitrogen at the time of remission. In 12 patients the marrow cells were fractionated using discontinuous albumin gradients in an attempt to separate normal cells from residual leukemic cells. Twelve patients achieved complete remission (CR); in 9 additional patients signs of engraftment were evident but death occurred before achievement of CR. Seven of 12 AML patients, which were treated with bone marrow transplantation as first treatment of their relapse, achieved CR. Four of 5 patients with ALL, whose bone marrows were collected during first remission, reached CR. The median CR duration was 4+ months and the median survival of the patients reaching CR was 6+ months. Autologous bone marrow transplantation offers a good chance of CR (66%), when marrow is collected during first remission and used as first treatment for AML in third relapse and ALL in second relapse.


Stem cell kinetics in humans: studies during autologous nonfrozen bone marrow transplantation.

Multiple blood and bone marrow determinations of CFU-C were carried out during autologous bone marrow infusion in four patients treated with high dose chemotherapy. A rise in blood CFU-C was seen during the infusion while bone marrow CFU-C increased after several hours.


Myelopoietic enhancement by immunoadjuvants: in vitro studies for their rational use in neutropenic patients.

Following marrow transplantation, the clinical course of the patients is invariably complicated by severe infections due to neutropenia and immuno-incompetence. With a view to explore means of alleviating these complications, Bacillus Calmette-Guerin (BCG), methanol extraction residue of BCG (MER), Corynebacterium parvum (C. parvum) and Pyran were examined for their myelopoietic activity on human marrow. Light density (less than or equal to 1.077 g/ml) unfractionated (UF) cells (2 X 10(6)/ml and 2ml/dish), and adherent (Ad) and non-adherent (N-Ad) cells alone (derived from 4 X 10(6) UF cells per dish) were incubated with and without the above agents. The conditioned media (CM) were harvested at 24, 72, 96, and 168 hours and colony stimulating activity (CSA) assayed against light density non-adherent human marrow cells using double layer agar-culture system. CSA was maximum at 72 and 96 hours followed by an invariable decline at 168 hours' incubation. The optimal concentrations releasing maximum CSA varied for each agent. CM prepared with higher concentrations were less active. CM prepared in the presence of Pyran had no CSA. Comparisons of the maximally released CSA by the optimal concentrations of these agents revealed BCG and MER to be the most active. The CSA elaborated by UF cells was more than the total combined activity from separately incubated Ad and N-Ad cells. Re-addition studies revealed that almost full CSA could be recovered when Ad and N-Ad cells were incubated together at a ratio of 1:3. Different immunoadjuvants have a differing capacity to elaborate CSA from human marrow cells and each has an optimal concentration for maximum CSA release. This may require a cell-cell and/or humoral interaction(s) between Ad and N-Ad cells.


Antigenic characteristics of normal and malignant human hematopoietic cells. Potential use of antigenic differences in cell separation and marrow transplantation.

Recent studies using cytotoxic and cell separation techniques have identified differentiation-related antigens on human hematopoietic cells. These results, combined with bone marrow transplantation studies, have yielded a picture of antigenic modulation from the pluripotent hematopoietic stem cell through the differentiated end cells.


Recovery of CFU-C after freezing of normal and leukemic human bone marrow.

Studies have been carried out to determine the viability of leukemic and normal human bone marrow, cryopreserved in liquid nitrogen at -196 degrees C, using changes in total cell numbers and granulocyte colony forming ability in vitro. These studies have shown that there is considerable variability in the recovery of CFU-C from individual specimens. When the overall recovery, in all patients, is taken into account, there is a gradual decline in CFU-C numbers to about 60% after 24 months of freezing. CFU-C recovery is closely correlated with recovery of total cell numbers.


Cell biological effects of total body irradiation on growth and differentiation of acute myelogenous leukemia cells compared to normal bone marrow.

Radiation therapy is used as total body treatment in preparation of the acute myelogenous leukemia (AML) patient for bone marrow transplantation. Many AML patients will have residual leukemia cells at the time of total body irradiation (TBI). In the present study, the effect of TBI on leukemic myeloid cells was compared to the effect on normal marrow granulocytic stem cells (CFUc) in vitro. Little difference from that of normal CFUc was found in the radiosensitivity of two mouse myeloid leukemia cell lines. The effect of TBI on growth of WEHI-3 or J774 cells in millipore diffusion chambers was stimulatory. These AML cell lines as well as others derived from Friend or Abelson virus infected in vitro long term mouse marrow cultures showed some morphologic differentiation by 7 days growth in diffusion chambers in irradiated heterologous rat hosts, but immature cells predominated by day 21. Thus, evidence in murine models of AML indicates that residual AML cells surviving chemotherapy will show no greater susceptibility to radiation killing compared to normal stem cells and will rapidly repopulate the irradiated host.


The critical influence of timing of combined modality cytoreductive regimens.

When sequential high dose drug-radiation cytoreductive regimens are employed, the interval between chemotherapy and total body irradiation (TBI) can markedly influence both toxicity in rapid cell renewal systems and the probability of successfully ablating normal and malignant hemopoietic stem cells. This is primarily due to the intense regenerative responses manifested by cells of these tissues that survive the first cytoreductive insult. Since the total doses of chemotherapy and radiation used are limited by toxicity to slowly or non-proliferating tissues, the best therapeutic effect should be obtained when the interval between chemotherapy and TBI is the shortest consistent with acceptable toxicity to rapid cell renewal systems, provided adequate time for drug metabolism and excretion is allowed to minimize the risk of interactive toxicity. In mice treated with piperazinedione 50 mg/m2 and fractionated TBI, the optimal interval between drug administration and irradiation is 3-4 days.


Accelerated hemopoietic recovery following the infusion of cryopreserved autologous bone marrow in humans.

High dose combination chemotherapy was given to 22 patients with malignant lymphoma resistant to conventional chemotherapy. This was followed in 12 patients by an infusion of their cryopreserved autologous bone marrow; 10 patients received chemotherapy alone and serve as controls. Patients receiving marrow recovered leukocyte, granulocyte, and platelet function significantly faster than controls demonstrating that cryopreserved autologous bone marrow infusions accelerate hemopoietic recovery. Four patients with Burkitt's lymphoma resistant to conventional chemotherapy appear cured of their disease following this single treatment with higher doses of chemotherapy.


Autologous marrow transplantation for patients with acute myelogenous leukemia--a preliminary report.

Twelve consecutive adult patients with acute myelogenous leukemia have been entered on a treatment protocol which examines the role of "remission-intensification" during maintenance using high-dose chemoradiotherapy and autologous remission marrow transplantation. Nine patients have achieved complete remission: 5/9 patients have had remission marrow stored followed by high-dose chemoradiotherapy and autologous marrow transfusion; two patients were removed from study because of excessive toxicity during remission-induction precluding high-dose therapy; two patients are currently ready for marrow storage; and two patients are receiving remission-induction therapy. Of the five transplanted patients, four experienced excellent return of blood counts and one patient has had prolonged pancytopenia and continues to require red cell and platelet transfusions. There have been no serious infectious or hemorrhagic problems associated with post-transplant period in any of the patients. Autologous remission bone marrow transplantation following lethal high-dose chemoradiotherapy results in effective restoration of normal hemopoiesis, is associated with acceptable toxicity and may be an effective means of increasing the numbers of acute leukemia patients having long-term complete remission.


Autologous bone marrow transplantation in the treatment of selected human malignancies: The Johns Hopkins Oncology Center Program.

Preliminary clinical trials using cryopreserved autologous bone marrow reinfusion have now been carried out at our institution in 5 children and 2 adults with advanced stages of neuroblastoma, rhabdomyosarcoma, non-Hodgkin's lymphoma and small cell carcinoma of the lung. Normal numbers of in vitro colony forming cells (CFU-C) were obtained from these patients despite prior courses of combination chemotherapy. The dose of marrow cells cryopreserved ranged from 1-6 X 10(8) cells/kg and recovery of CFU-C after thawing averaged 50%. Partial or complete hematologic reconstitution was achieved in all patients. The time for recovery ranged from 10-43 days for leukocytes (greater than 1000 cells/mm3) and 23-45 days for platelets (greater than 50,000/mm3). Two patients have died of interstitial pneumonitis due to cytomegalovirus. Three patients have died of recurrent tumor at 40, 48 and 156 days post-transplant. Two patients have had significant therapeutic benefit. One of these had a stable partial response permitting the use of further post-transplant therapy and is alive and well 16+ months post-transplant. The other patient had a complete response and remains tumor-free 25+ months following therapy. We conclude: 1) Autologous bone marrow reinfusion permits hematologic reconstitution following marrow-ablative therapy. 2) A quantity of marrow sufficient for this purpose remains viable following cryopreservation even when obtained from patients previously exposed to chemotherapy. 3) Autologous bone marrow reinfusion now allows the exploration of more intensive cytoreductive therapy in selected malignancies.


Autologous bone-marrow and peripheral blood buffy coat cell infusion in the treatment of chronic myeloid and acute leukemia.

Autologous marrow infusion has been attempted in three patients with chronic myeloid leukemia (two in blast crisis, one with severe myelofibrosis and pancytopenia) and one patient with acute lymphatic leukemia. One patient with blast crisis of CML expired prior to marrow infusion. One patient with myelofibrotic phase of CML is alive seven months post marrow infusion. The other two patients expired 6 and 16 days post marrow infusion. Bone-marrow repopulation is feasible in the face of severe myelofibrosis.


Autologous bone marrow transplantation in patients with cancer.

Ten patients with non-leukemic neoplasms received intensive, marrow-lethal doses of drugs and radiation followed by rescue with autologous cryopreserved bone marrow (nine) or marrow from an identical twin as part of a phase 1-2 study. Nine patients had extensive disease that was unresponsive to conventional therapy. Marrow engraftment was documented in all evaluable cases and most patients had a substantial anti-tumor response. Three patients are alive from 4 to 10 months following transplantation without evidence of disease.


High dose melphalan and non-cryopreserved autologous bone marrow treatment of malignant melanoma and neuroblastoma.

Autologous non-cryopreserved bone marrow infused 8 hours after an intravenous injection of melphalan, 140 mg/m2, accelerates bone marrow recovery. This effect is most noticeable in the recovery of peripheral blood granulocytes. Twenty patients with disseminated malignant melanoma were treated with this regimen: there were 12 responses, two of them complete but the toxicity of the treatment was not sufficient to justify using this method of treatment routinely since survival was little influenced by treatment (4-11 months). In 8 patients with disseminated neuroblastoma, high dose melphalan/autograft was used in a program of combined modality treatment. Three of the patients are disease free at 16, 11 and 6 months and in one the disease is 'static', not having grown for 13 months. The treatment for this tumour deserves further exploration, and perhaps similar treatment ought to be explored for other tumours.


Intensive 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) autologous bone marrow transplantation therapy of refractory cancer: a preliminary report.

Eighteen patients with refractory malignancies were treated with escalating doses of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and autologous bone marrow transplants (BMTX). Hematopoietic recovery was similar with doses of 300mg/m2qd X 3 and 500mg/m2qd X 3 providing suggestive evidence of a myeloprotective effect of the BMTX. Extramedullary toxicity was sporadic but occasionally severe; pulmonary fibrosis and severe cholestatic jaundice were seen in one case each. Antitumor responses were noted in patients with brain tumors, melanoma, hematological neoplasms and lung cancer.


High dose chemotherapy with autologous bone marrow transfusion.

Details are given of response and toxicity after high dose chemotherapy followed by autologous bone marrow infusion. High dose nitrosurea therapy (600-1000 mg/m2 BCNU) was predominantly associated with hematopoietic toxicity but recovery was within 4-5 weeks after BCNU therapy. High dose combination chemotherapy using cytoxan, VP-16-23 and +/- BCNU produced a response rate of approximately 80% (CR + PR + less than PR) of usually short duration in 18 evaluable patients, 17 of whom were previously treated. Hematopoietic recovery was usually complete in 4 weeks. Initial experience with the hematopoietic toxicity experienced after high dose mitomycin and ABMT is also detailed. Future proposals utilizing high dose chemotherapy with ABMT in selected tumors is presented with associated rationale.


Clinical results of autotransplantation after nitrosourea therapy.

Bone marrow harvested from two previously treated patients was cryopreserved prior to their first dose of Methyl CCNU. The thawed bone marrow was reinfused 96 hours after a subsequent dose of MeCCNU in one patient after the previous dose caused severe myelosuppression. In the other patient, the bone marrow was given 96 hours after an escalation of the MeCCNU dose. In both patients, myelosuppression was abrogated and viable stem cells were recovered in the peripheral blood.


Modulation of hematopoiesis and survival after high-dose chemotherapy or radiotherapy: review and discussion of possible mechanisms.

This paper reviews ways by which growth of transplanted or surviving hematopoietic stem cells might be enhanced to improve survival in the case of autologous marrow transplantation. Most of the treatments known to have such effects have been used in isologous mouse models and improve animal survival by enhancing hematopoietic recovery after high doses of whole body irradiation or chemotherapy. Although some of these were studied 20 years ago, the use of such treatments in man has awaited the realization that some disseminated cancer can only be eliminated by use of various treatments where hematopoietic damage is the limiting factor. Although increasing the number of transplanted pluripotent stem cells is the most certain way to hasten return of needed blood cells in transplant patients, several of the treatments listed in the paper have growth enhancing effects on both surviving host stem cells and transplanted cells. There are recent studies which indicate that such priming treatments may also be effective in other normal tissues such as gut and bladder epithelium. At least one study in man has intentionally applied this approach with the expected benefit and others may have done so inadvertently. Much work remains to find new combinations to select the best priming treatments and intervals in man and to determine whether or not such treatments are effective against tumors.


Hemopoietic reconstitution following autologous bone marrow and peripheral blood mononuclear cell infusions.

We have undertaken a series of experiments using a canine model to determine the minimal number of cryopreserved autologous bone marrow or peripheral blood mononuclear cells needed to protect animals from otherwise lethal total body irradiation. We further have compared the kinetics of engraftment using hemopoietic cells from these two sources. Animals engrafted with bone marrow required .25 X 10(8) nucleated bone marrow cells/kg to protect them; those engrafted with peripheral blood mononuclear cells required 6.0 X 10(8) cells/kg. Myeloid and platelet recovery appeared more rapid after bone marrow infusion while lymphoid recovery was more rapid when peripheral blood mononuclear cells were used.


Liquid storage of bone marrow.

A program of nonfrozen autologous bone marrow rescue was used to treat patients with refractory nonhematological neoplasms. In vitro storage of bone marrow cells and CFU-C suggests that storage at 4 degrees or 10 degrees for periods up to five days can be undertaken with only minor loss of CFU-C. However, since the loss occurred in a gradual fashion, shorter duration of storage would probably be better. Hematological recovery after high dose chemotherapy occurred faster in patients given more CFU-C and those with shortest duration of storage prior to reinfusion.


Psychological issues in bone marrow transplantation.

We studied the psychological and emotional problems experienced by seven children and their families who underwent bone marrow transplantation at the University of Colorado Medical Center from 1973 to 1975. These problems included (1) anxiety and depression relating to isolation, fear of death, and painful procedures; (2) an overdependence associated with a feeling of helplessness; (3) anger directed toward both the staff and the parents; (4) a reduced tolerance for medical procedures; and (5) periodic refusal to cooperate. Initially we had been concerned that patients might become agitated, psychotic, or even suicidal. These did not occur. Severe anxiety over bodily changes was not a problem. We did not encounter prolonged refusal to cooperate, refusal to remain in isolation, or drug addiction. Important aspects in management included an honest, straightforward, and direct discussion of all aspects of transplantation, including the potential complications and the risks of death from the underlying disease or from complications of transplantation. A firm but understanding approach to the patients appeared to be the most effective method to develop their continuing cooperation. The opportunity for patients to express verbally their fears of procedures and of death was essential. The donors needed help in working through their feelings of guilt if a transplant was not successful. The parents needed continuing psychological support for the many personal, social, and psychological difficulties which they had to face.


A large form of renin from normal human kidney.

The apparent molecular size of circulating human plasma renin is 43,000 daltons. In this report, the Stokes' radius of renin extracted from human kidney cortex in low ionic strength buffer in the presence of protease inhibitors was shown to correspond to an apparent molecular weight of 58,000 +/- 3,000. If protease inhibitors are omitted, if the kidney tissue is frozen and thawed multiple times, or if the kidney extract is acidified to pH 2.8, renin activity of an apparent molecular size of 42,000 +/- 3,000 can be identified. Both species of renin bind to an affinity support bearing the substrate analog inhibitor His-Pro-Phe-His-Leu-D-Leu-Val-Tyr. Antibody raised to the higher molecular form of the enzyme inhibits the activity of both forms in an equivalent manner. These observations suggest that the larger form of renin may be a biosynthetic precursor of plasma renin, either in the form of a zymogen or an enzyme-binding protein complex.


A specific radioimmunoassay for human beta-lipotropin.

A homologous RIA for human beta-lipotropin (beta hLPH) has been developed. At a final dilution of 1:24,000, the antiserum employed shows cross-reaction with beta hLPH but none with human beta-MSH (beta hMSH), and it is concluded that the antigenic determinant lies within the N-terminal 1-36 region of beta hLPH. With extraction of 3-ml plasma samples, the assay is sufficiently sensitive to measure circulating beta hLPH levels in normal individuals at 0900 h (25-200 pg/ml). There is a circadian variation with levels falling to (less than 20-80 pg/ml) at 2300 h. beta hLPH levels rise after metyrapone and after insulin-induced hypoglycemia, and fall after administration of dexamethasone. In patients with a variety of diseases of the pituitary-adrenal axis, levels of beta hLPH follow immunoreactive ACTH levels, although the two are not always secreted on a 1:1 molar basis.


Increased insulin binding to monocytes after acute exercise in normal man.

The effect of exercise on 125I-insulin binding to monocytes was studied in 10 healthy subjects. We found that acute exercise (bicycle ergometer for 3 h) led to a significant increase (36%) in insulin binding to monocytes, which returned to basal levels by 24 h after exercise. The changes in insulin binding were mainly due to an increase in receptor affinity. The increase in insulin binding during exercise correlated with the fall in plasma glucose but not with the decrease in plasma insulin or increase in blood ketones. We conclude that acute exercise increases insulin binding which may contribute to augmented insulin sensitivity associated with exercise.


Glucose-lowering effect of insulin by different routes in obese and lean nonketotic diabetic patients.

The absorption of insulin and its glucose-lowering effect were compared after the administration of crystalline insulin by sc, im, and iv routes in 29 obese and 10 lean nonketotic diabetic patients, none of whom had consciously received insulin previously. Each of the patients received insulin in a dose of 0.1 U/kg BW by the im, sc, and iv routes in a randomized fashion on 3 different days. Plasma glucose, immunoreactive insulin (IRI), and immunoreactive glucagon (IRG) were measured at intervals over the first 4 h. The t1/2 (mean +/- SEM) after iv administration of insulin in obese and lean diabetics was, respectively, 5.3 +/- 0.2 and 4.8 +/- 0.4 min; these were not significantly different. Intravenous injection produced its highest level of IRI in 2 min in both groups. Thereafter, a rapid drop was observed with return to the basal level by 90 min. Equivalent amounts of im and sc insulin produced a maximal increase in plasma IRI at 60 min in both groups. Plasma IRI after iv insulin injection was significantly higher than after sc and im insulin injections at 10 and 20 min (P less than 0.001) and significantly lower than the im and sc groups at 60, 90, 120, 150, 180, 210, and 240 min (P less than 0.001). After iv insulin, plasma glucose at 30, 40, 50, and 60 min was significantly lower than after im and sc insulin (P less than 0.001), but over the 4-h study period, the glucose-lowering effect and the area under the curves for glucose response to IRI by the three routes were the same in both lean and obese diabetic subjects. The mean basal IRI in lean patients was 18 +/- 4 microU/ml, which was significantly lower (P less than 0.05) than in obese patients (26 +/- 2 microU/ml). No significant difference was observed in fasting IRG in lean (96 +/- 12 pg/ml) vs. obese (108 +/- 10 pg/ml) patients. No significant increase in IRG was noted with equivalent amounts of sc, im, and iv injection in the lean and obese patients. These studies demonstrated that although iv injection of insulin produces a more rapid initial decline in plasma glucose, the overall glucose-lowering effect by insulin given iv, im or sc is similar in nonketotic lean or obese diabetic subjects.


Studies of plasma aldosterone in anephric people: evidence for the fundamental role of the renin system in maintaining aldosterone secretion.

When plasma aldosterone (PA) was measured in 32 predialysis samples from 18 anephric subjects more than 3 months postnephrectomy, all but two values were subnormal, 16 measurements were undetectable, i.e. below 0.35 ng/100 ml, and PA averaged only 1.7 ng/100 ml +/- 0.5 (SE) in the remaining 16. In contrast, PA averaged 29.7 +/- 5.6 ng/100 ml in 12 dialysis patients with kidneys. Plasma renin activity was undetectable in 15 assays of anephric subjects and averaged 0.5 +/- 0.1 ng/ml/h in the remaining 17. However, these latter measurements appeared to be an artifact caused by the presence of plasma prorenin. Serum potassium fell during dialysis and appeared responsible for the concurrent 75% fall in PA observed in six nephric patients (delta 14 ng/100 ml), and in three anephric patients (delta 3.3 ng/100 ml) with low but measurable PA values. Overt hyperkalemia was required for potassium to stimulate aldosterone into the normal range in anephric subjects. The angiotensin II analog, saralasin, also increased PA slightly in five studies. Renin and PA fell after nephrectomy and increased after transplantation. Decreases and increases in renin always preceded those of aldosterone. Usually it took several weeks for aldosterone to fall to undetectable levels and several days to return to normal levels after renal transplantation. These observations suggest that the baseline plasma level of angiotensin determines both the baseline level of aldosterone secretion and its capacity to respond either to more angiotensin or to other stimuli. In the absence of renin, the adrenal almost loses its capacity to generate aldosterone. The data support the view that the renin system plays a fundamental role in maintaining aldosterone secretion in man.


Effects of antiinsulin receptor autoantibody on the metabolism of rat adipocytes.

We have studied the effects of the serum from a patient with an unusual form of diabetic syndrome with extreme insulin resistance on the metabolism of rat adipocytes in vitro. This serum and IgG fractions from it inhibited the [125I]insulin binding to isolated adipocytes and stimulated the 2-deoxyglucose uptake, glucose oxidation, and the incorporation of amino acids into protein. In addition, these fractions inhibited the lipolysis induced by beta 1-24 ACTH in isolated adipocytes. The insulin-like effects of this serum and the effects of insulin were not additive at their maximal concentrations. The inhibition of [125I]insulin binding was due to a decrease in receptor affinity rather than to a change in receptor number by Scatchard plot analysis. Both the inhibition of insulin binding and the insulin-like effects on rat adipocytes were neutralized by antihuman IgG. In addition, these insulin-like effects were abolished by trypsin treatment of adipocytes. These facts suggest that this serum has a circulating antibody directed at or near the insulin receptor itself and that this antibody mimics the insulin effect on rat adipocytes by binding to the insulin receptor in vitro.


Presence of insulin-renin-aldosterone-potassium interrelationship in normal subjects, disrupted in chronic hemodialysis patients.

The factors regulating aldosterone secretion in normals and in patients on chronic hemodialysis were studied by the determination of the circadian rhythm of plasma aldosterone, renin, cortisol, insulin, potassium, sodium, and glucose. Four normal volunteers and eight normotensive patients on regular dialysis treatment (RDT) were studied during prolonged recumbency on low sodium diet. A definite circadian rhythm for renin could not be demonstrated in RDT patients. The significant simple and multiple correlation coefficients found in normal subjects suggest that insulin participates in the regulation of aldosterone together with the other known factors: ACTH, renin, and potassium. In chronic renal failure, however, when basal conditions were maintained during prolonged recumbency, the correlations of insulin and renin with aldosterone were not found, suggesting that in this condition aldosterone secretion is controlled by ACTH and potassium. As a direct influence of insulin on aldosterone could not be demonstrated by multiple variance analysis, it seems that insulin is related to aldosterone indirectly through renin and/or potassium. The presence of significant correlations between insulin-potassium and potassium-aldosterone in RDT patients, without a significant insulin-aldosterone correlation, suggest that in the normals insulin participates in aldosterone regulation through renin secretion and not through potassium. A correlation between potassium and renin was not found in normals or in RDT patients.


Normal pregnancies after treatment of hyperprolactinemia with bromoergocryptine, despite suspected pituitary tumors.

Bromocryptine treatment was administered to 15 patients with amenorrhea and galactorrhea (AG) and to 1 patient with amenorrhea. All of them had increased plasma PRL levels. Of these 16 patients, 4 had a normal sella turcica (ST; group STO), 4 had a slight enlargement (group ST+), and 7 had a clear enlargement of ST (ST++) but no evidence of suprasellar extension. Ovulation was restored in 15 patients by bromocryptine treatment only. In one patient, ovulation resumed only after human pituitary gonadotropin treatment in combination with bromocryptine. There was no correlation between basal prolactinemia, PRL stimulability or suppressibility, the size of ST, or the efficiency of bromocryptine treatment. Every patient with normal LH response to either LRH or clomiphene or both resumed ovulation. Ovulation resumed in 3 patients among the 4 with abnormal LH response to either LRH or clomiphene or both. Among the 14 who desired pregnancy, 13 became pregnant. To date, 12 patients (ST++, 5; ST+, 3; STO, 4) have delivered normal babies. The courses of pregnancy were normal. During pregnancy, no change of ST was noted on lateral and frontal skull x-ray performed in every patient at trimonthly intervals. There was no change in the sellar index in 10 patients after pregnancy, as compared to the pretreatment status. In the presence of a pituitary adenoma or in patients with hyperprolactinemia and amenorrhea and galactorrhea, bromocryptine treatment may cure sterility without pituitary complication during pregnancy.


Stimulation of pancreatic polypeptide and glucagon secretion by 2-deoxy-D-glucose in man: evidence for cholinergic mediation.

Hypoglycemia is known to stimulate human pancreatic polypeptide (hPP) secretion. To explore further the relationship between glucose availability and hPP release, we have examined the effect of tissue glucopenia induced by 2-deoxy-D-glucose (2-DG) on hPP plasma levels in normal subjects. As this glucose analogue activates the autonomic nervous system, we have also studied the influence of prior atropinization upon the hPP response to 2-DG. Moreover, we have tested the effects of iv epinephrine and norepinephrine on plasma hPP concentrations. Circulating glucagon was also measured. After the iv infusion of 2-DG (50 mg/kg), plasma hPP increased steeply from a fasting value of 104 +/- 24 pg/ml (SEM) to a peak of 2175 +/- 639 pg/ml at 45 min (P less than 0.01) and remained significantly elevated throughout the test. In contrast, prior injection of atropine (1 mg iv) lowered basal hPP levels and reduced conspicuously the hPP response to 2-DG. Epinephrine administration (6 micrograms/min for 60 min) did not significantly modify plasma hPP concentrations. However, 2 h after epinephrine withdrawal, circulating hPP showed a brisk elevation coinciding with the decline of glycemia to subbaseline values. During norepinephrine infusion (6 micrograms/min for 60 min), only a minor and transient increase of plasma hPP was found. Plasma glucagon rose significantly after 2-DG infusion, but this response was virtually absent in the atropine experiment. Whereas the well known glucagon tropic activity of epinephrine was evidenced, norepinephrine failed to exert an obvious effect on glucagonemia. Our data demonstrate that 2-DG induces a powerful stimulation of hPP secretion in normal subjects and suggest that this action is mediated in part, if not entirely, by the parasympathetic nervous system. On the other hand, a major role of the sympathoadrenal system in response of hPP to 2-DG or to hypoglycemia does not seem probable. Finally, the hyperglucagonemic effect of 2-DG seems also to be dependent on cholinergic transmission.


Diurnal variation in insulin binding to human monocytes.

We studied insulin binding to monocytes isolated from dieting and fasting young healthy persons during a 24-h period. In the persons who ate their usual food, we found a characteristic diurnal variation in the cellular insulin binding. Daytime insulin binding was low with a minimum in the afternoon; binding increased during the evening, reaching a maximum in the early morning. This variation seemed to be diet related, as total fasting abolished the circadian rhythm. Changes in cellular insulin binding were due to fluctuations in the binding affinity rather than alterations in receptor concentration. The circadian variation in insulin binding was grossly parallel to the well known variations in insulin sensitivity and glucose tolerance during the day.


Induction of growth hormone and prolactin secretion by luteinizing hormone-releasing hormone and its blockade by bromoergocriptine in acromegalic patients.

In an attempt to evaluate the effect of LRH on GH and PRL secretion, LRH in a dose of 100 micrograms was injected iv to 22 patients with active acromegaly. A definite increase in both serum GH and PRL levels was observed in one patient, while four other subjects responded to LRH to secret either GH or PRL. The effectiveness of LRH on GH and PRL release was not correlated with the age or sex of patients, the basal levels of serum GH or PRL, the GH response to TRH administration, or the increase in serum gonadotropin concentrations after LRH injection. In three patients, this LRH-induced GH and PRL release was completely blocked by pretreatment with 2-bromo-alpha-ergocriptine (CB154). The results suggest that elevations of GH and PRL after LRH occur rather sporadically in acromegalic patients and that the action of LRH on GH release is occasioned by a different mechanism from that of TRH.


Demonstration by immunoperoxidase histochemistry of calcitonin in the anterior lobe of the rat pituitary.

We have demonstrated by a specific immunoperoxidase procedure the presence of calcitonin-containing cells in the rat pituitary gland. These cells are widely distributed throughout the anterior lobe and seem to constitute the entire population of cells of the intermediate lobe. No such cells were seen in the posterior lobe. The presence of calcitonin-containing cells in the pituitary provides novel implications about the physiological significance of this hormone.


Direct effect of insulin on secretion of insulin, glucagon, gastric inhibitory polypeptide, and gastrin during maintenance of normoglycemia.

The direct effect of insulin on the secretion of insulin (as measured by C-peptide), glucagon, gastric inhibitory polypeptide, and gastrin was studied in normal subjects by infusing insulin while the plasma level of glucose was maintained in the normal fasting range (euglycemic clamp). Insulin-induced hypoglycemia resulted in increases in circulating glucagon and gastric inhibitory polypeptide, a decrease in C-peptide, and no change in gastrin levels. In contrast, during the euglycemic clamp, insulin was found to behave a direct suppressive effect on the secretion of glucagon, C-peptide, and gastrin, but no effect on levels of gastric inhibitory polypeptide.


Evidence for a defect in pituitary secretion of luteinizing hormone in chronic alcoholic men.

To characterize the defect in the hypothalamic-pituitary-gonadal axis of alcoholic men, acute and chronic LRF responses were evaluated in 22 chronic alcoholic men with varying degrees of biochemically and histologically confirmed liver disease. In addition, acute LRF responses in 14 normal men, before and at the end of 72 h of administration of 2 ml/kg/day 95% ethanol, were evaluated. The alcoholics hd significantly reduced basal testosterone and elevated gonadotropin levels (both FSH and LH) compared to the normal volunteers (P less than 0.02). Serum concentrations of estradiol and PRL did not differ between alcoholics and normal volunteers. A 100-micrograms bolus of LRF resulted in a 3-fold increase of LH in alcoholic men as compared to a 6-fold increase of serum LH in normal volunteers. No significant difference in the LRF-induced FSH responses was observed. When the response of normal volunteers to LRF before and after ethanol administration was evaluated, basal levels of both gonadotropins were increased after alcohol administration and a reduced LRF-induced LH response was observed. Based upon these results, we conclude that: 1) the central hypothalamic-pituitary defect known to exist for LH secretion is in part due to inadequate pituitary secretion and 2) acute alcohol ingestion in normal men suppresses the LRF-induced LH but not the FSH response.


Modulation of basal ketone body concentration by cortisol in diabetic man.

This study explored the relationship between metyrapone blockade of endogenous cortisol secretion and the plasma concentration of basal ketone bodies in insulin-dependent diabetic man. Endogenous cortisol secretion was reduced with metyrapone administration (as assessed by a reduction in plasma cortisol and an increase in deoxycortisol concentration) and then simulated with two different schedules of exogenous cortisol administration. Our results demonstrate that administration of metyrapone suppresses plasma cortisol concentration which may then be elevated by the oral ingestion of cortisol. The suppression of endogenous cortisol secretion with metyrapone results in a 50% reduction in basal plasma ketone body concentration. When endogenous cortisol secretion was simulated with the oral administration of 30 mg cortisol, plasma ketone body concentration returned to that concentration observed in the control study. When 60 mg cortisol were administered in divided dosages to the diabetic subjects, hyperketonemia resulted. These results suggest that the normal diurnal variation in plasma cortisol concentration may modulate the basal plasma ketone body concentration in diabetic man. The mechanism of this modulation may be a direct effect of cortisol or may be secondary to cortisol's lipolytic activity and/or its effects on elevating plasma glucagon concentration.


Gastric inhibitory polypeptide in acquired pancreatic diabetes: effects of insulin treatment.

Oral glucose tolerance tests were done in eight insulin-requiring pancreatic diabetic patients to study the effect of withdrawal of insulin treatment on gut hormone release. Basal levels of gastric inhibitory polypeptide (GIP), glucagon-like immunoreactivity, and immunoreactive glucagon levels rose on insulin withdrawal, more so in patients on short-acting insulin, and were lowered by insulin treatment. Insulin treatment did not affect the GIP, glucagon-like immunoreactivity, or IRG responses to oral glucose. Improved glucose tolerance was greater in patients receiving soluble insulin than in those receiving lente insulin, and there was a significant positive linear correlation between basal plasma GIP and blood glucose levels in these patients. Therefore, it is suggested that insulin treatment lowers basal hormones levels, possibly via a metabolic effect, whereas the hormone responses to oral glucose may be controlled by several factors unrelated to insulin administration or changes in glucose homeostasis.


Increased monocyte receptor binding of [125I]insulin in infants of gestational diabetic mothers.

Monocyte insulin receptor binding has been shown to be inversely correlated with basal insulin concentrations in a variety of clinical conditions and is believed to reflect autoregulation of receptor properties of insulin. We examined the insulin receptor in the infant of the gestational diabetic mother (IGDM) where hyperinsulinism has been implicated in the attendant metabolical abnormalities. Insulin concentrations in plasma of cord blood of IGDM were significantly greater than in normals delivered between 36-38 weeks by elective cesarean section (101.3 +/- 9.8 vs. 59.9 +/- 9.4 microU/ml; P less than 0.005). [125I]Insulin binding to receptors of monocytes obtained from cord blood showed that IGDM had more receptor sites per monocyte than normal adults and normal infants. In normal infants of similar gestational age, a significant correlation was found between birthweight and binding which was not observed in IGDM. Monocytes from both normal infants and IGDM showed greater affinity for insulin than those from adults. At plasma insulin concentrations of 1 and 4 ng/ml, monocytes from IGDM had about 10 times and normal infants had about 4 times as many sites occupied as those from normal adults. Monocytes from IGDM seem to develop increased concentrations of insulin receptors with hyperinsulinemia. Thus, despite increased ambient levels of insulin, monocytes of IGDM seem to develop increased concentrations of insulin receptor as well as increased affinity for the hormone.


Gastric inhibitory polypeptide (GIP) and insulin release in duodenal ulcer patients.

To investigate the role of gastric inhibitory polypeptide (GIP) in the hypersecretion of glucose-stimulated insulin release in duodenal ulcer disease, serum glucose, insulin, and immunoreactive GIP (IR-GIP) were measured in 18 healthy subjects and 10 duodenal ulcer patients after glucose ingestion. Although the serum glucose and insulin were significantly greater (P less than 0.05) at 15 and 60 min in the ulcer group, the total integrated glucose areas were similar (20,552 +/- 837 vs. 19,154 +/- 745 mg-min/ml). In contrast, the total integrated insulin area was significantly greater (P less than 0.05) in the ulcer patients (12,873 +/- 2,082 vs. 8,216 +/- 1,072 micro U-min/ml). Mean IR-GIP levels were significantly greater (P less than 0.05) in the ulcer group at 15-120 min of the study, as was the total integrated area (244,755 +/- 34,934 vs. 126,595 +/- 17,468 pg-min/ml). The exaggerated insulin release to oral glucose may be due to the synergistic action of higher blood glucose and greater IR-GIP release in this disease.


Insulin resistance, acanthosis nigricans, and normal insulin receptors in a young woman: evidence for a postreceptor defect.

We have previously described a group of young females with virilization, acanthosis nigricans, insulin resistance, and markedly decreased binding of insulin to its receptor (syndrome of insulin resistance and acanthosis nigricans type A). The present report concerns a 15-yr-old female with clinical features indistinguishable from the type A patients, including virilization, acanthosis nigricans, and extreme resistance to endogenous and exogenous insulin. Insulin levels were 400-650 microU/ml while fasting and were over 2200 microU/ml when stimulated. Proinsulin was less than 10% of the total immunoassayable insulin. In distinct contrast to the type A patients, insulin receptors on cells from this patient were entirely normal on the basis of specificity, negative cooperativity, affinity, concentration, and interaction with antiinsulin receptor antibodies. These findings suggest the presence of an intracellular defect as the cause of the observed insulin resistance.


Correlations between plasma renin activity, angiotensin II, and aldosterone.

To obtain more information about the relationships of the components of the renin-angiotensin-aldosterone system, we measured plasma renin activity (PRA) and the concentrations of plasma angiotensin II (AII) and aldosterone (PA) in 36 healthy 19-22-yr-old students, both after a night's bed rest and after 2 h of ambulation. The correlations of these parameters were calculated. PRA, AII, and PA were determined by RIAs. The values of AII after rest correlated positively both with PRA (r = 0.80; P less than 0.001) and with PA (r = 0.51; P less than 0.01). AII also showed a good correlation with PRA (r = 0.77; P less than 0.001) and with PA (r = 0.67; P less than 0.01) after ambulation. The latter correlation was higher than that after rest. Positive correlations were also found between the increases in PRA and AII and between AII and PA. Further, a multiple regression analysis was carried out to calculate the regression equations for these parameters. The results support the view that PRA and AII parallel each other in normal physiological circumstances. They also seem to indicate that AII has an important role in the basal as well as in the posture-stimulated secretion of aldosterone in normal man.


Insulin receptors in human endothelial cells: identification and characterization.

Specific binding of 125I-insulin was found in cultured human endothelial cells obtained from human umbilical veins. The binding reaction was rapid and reversible, demonstrated receptor site-site interactions of the negatively cooperative type, and was dependent on the temperature, pH and duration of incubation. At 21 degrees C, steady-state conditions of binding occurred in 90 minutes, the pH optimum was 7.8 and less than 10% of the labeled hormone was degraded. Binding of tracer amounts of 125I-insulin was inhibited by concentrations of unlabeled insulin as low as 0.2 ng/ml and 50% inhibition was obtained at 2-5 ng/ml of unlabeled insulin. Unlabeled porcine insulin, porcine proinsulin and desoctapeptide insulin inhibited the binding of 125I-porcine insulin in direct proportion to their biological potencies, whereas to the cells was inhibited by antibodies against insulin receptors. We conclude that human endothelial cells possess specific receptors for insulin whose physio-chemical properties are similar to those of insulin receptors in other tissues.


Adrenal responses to pharmacological interruption of the renin-angiotensin system in sodium-restricted normal man.

We assessed the role of the renin-angiotensin system in the control of aldosterone secretion in response to sodium restriction in 62 normal subjects. Both saralasin, an angiotensin II antagonist, and SQ 20881, a converting enzyme inhibitor, induced a dose-related decrease in plasma aldosterone levels when the renin-angiotensin system was activated by restriction of sodium intake. Two types of experiments were performed with saralasin. In the first set, a dose-response relationship was established 20 min after beginning infusions ranging from 0.03-1.0 microgram/kg/min. The optimal dose was 0.1 microgram/kg/min, with a reduction in aldosterone levels of -10.1 +/- 3.8 ng/dl (P less than 0.025). Higher doses induced smaller reductions in aldosterone levels. In the second set, a 3-h infusion was given. The results were qualitatively similar but the magnitude was greater (-15 +/- 4 ng/dl; P less than 0.01). The aldosterone response 20 min after administration of SQ 20881 paralleled the angiotensin II response, with the first significant decrement (-6.5 +/- 1.5 ng/dl; P less than 0.01) occurring at 0.1 mg/kg and maximum (-10 +/- 3 ng/dl) occurring at 0.3 mg/kg. Thus, both agents produced qualitatively similar changes in aldosterone secretion in sodium-restricted normal subjects. However, neither reduced sodium restricted aldosterone levels to that measured in sodium-loaded subjects because of the intrinsic limitation of each agent. Saralasin is a partial agonist. SQ 20881 induces an increase in plasma renin activity via interruption of the short feedback loop, which probably limits its action. Yet, these data do support the hypothesis that angiotensin mediates the adrenal's response to sodium restriction in normal man.


Degradation and secretion of insulin in hepatic cirrhosis.

To clarify the mechanism of hyperinsulinism of hepatic cirrhosis, plasma insulin and C-peptide levels before and after oral glucose loads were measured in 34 patients with cirrhosis, 15 patients with chronic hepatitis, and 25 normal subjects. While plasma immunoreactive insulin (IRI) levels during oral glucose tolerance testing (OGTT) were significantly increased in cirrhotics, plasma immunoreactive C-peptide (CPR) levels were elevated slightly. The C-peptide to insulin ratio throughout OGTT was significantly smaller in cirrhotics than in normal subjects (P less than 0.01). A decreased hepatic insulin degradation rate has been suggested to one of the main causes of hyperinsulinism in hepatic cirrhosis. The ratio of the difference between basal and 30-min CPR values and basal and 30-min OGTT blood glucose values [delta CPR: delta BS(30)'] as well as the  delta IRI: delta BS(30') ratio was significantly decreased in cirrhotics (P less than 0.01). These results indicate that insulin secretion in response to a glycemic stimulus is reduced in cirrhotics. Both the ratios of the sums of six IRS and CPR values of OGTT (sigma CPR: sigma IRI) and delta CPR: delta BS(30') and sigma CPR: sigma BS(30') were found in inverse relationship with indocyanine green retention rate in cirrhotics.


Diurnal 18-hydroxy-11-deoxycorticosterone pattern in human stable hypertension.

Diurnal 18-hydroxy-11-deoxycorticosterone (18-OH-DOC) pattern was studied with RIA technique in 33 hypertensive patients in supine position and on normal sodium diet. The compound was evaluated every 2 h from 0800-2000 h. Simultaneously, plasma aldosterone and cortisol were measured. Abnormal 18-OH-DOC behavior was observed in only 2 out of 4 patients with Cushing's disease, while sporadic and slight elevations, synchronous with F, were seen in 5 out of 24 stable essential hypertensive patients [1 with normal plasma renin activity (PRA), 1 with low PRA, and 3 with high PRA]. 18-OH-DOC was normal in 2 cases of hypertension due to renal artery stenosis, in 1 patient with nephrosclerosis, and in 1 patient with horseshoe kidney. From these results, 18-OH-DOC does not seem to play an important pathogenetic role in stable essential hypertension, considering also the low mineralocorticoid activity of the compound.


Serum dopamine-beta-hydroxylase activity does not reflect sympathetic activation during hypoglycemia.

The relationship between serum dopamine-beta-hydroxylase (DBH) activity and sympathetic nervous system activation in response to insulin-induced hypoglycemia was investigated in healthy young male and female volunteers. Serum DBH activity and glucose levels were measured in blood samples obtained 30 min before and 90 min after the iv administration of regular insulin or placebo. All subjects developed significant hypoglycemia in response to the insulin and manifested signs of sympathetic activation, including increased heart rate, diaphoresis, and lightheadedness. However, despite obvious clinical manifestations of increased sympathetic activity, none of the subjects exhibited a significant alteration in serum DBH activity either in comparison to baseline or placebo values. These data question the suitability of serum DBH measurements for assessing adrenomedullary function in man.


Release of immunoassayable prostaglandin E by the human ischemic kidney.

Immunoassayable prostaglandin E concentration in renal venous plasma was measured in eight patients with renovascular hypertension. Two subjects with bilateral renal artery stenosis had similar concentrations from both renal veins. The six subjects with unilateral artery stenosis had greater concentration on the stenotic side in each case, suggesting that the human ischemic kidney produces increased amounts of prostaglandins.


Intestinal nutrient influence on the enteroinsular axis.

The gastrointestinal contribution to carbohydrate metabolism includes carbohydrate absorption and the release of gastrointestinal hormones that interact with the endocrine pancreas. To learn the contributions to the enteroinsular axis from different levels of the gastrointestinal tract and different nutrients in chyme, we determined serum concentrations of glucose, gastric inhibitory peptide (GIP), insulin, and glucagon postprandially in six normal subjects who underwent diversion of chyme just proximal to an occlusive balloon at the ligament of Treitz and jejunal infusion of saline or chyme carbohydrate, protein, and lipid, separately or in combination. Postprandial elevations of serum glucose, GIP, and insulin and decrease of serum glucagon were elicited predominantly from the bowel and its contents distal to the ligament of Treitz. In this segment, each chyme nutrient (but especially carbohydrate) significantly stimulated factors affecting carbohydrate metabolism. Protein and lipid were able to block carbohydrate-induced glucagon inhibition. The gastroduodenal segment, although containing several proposed insulinotropic hormones (gastrin, secretin, and cholecystokinin), had no effect on serum glucose of glucagon and stimulated only small insulin and GIP responses.


Hyperglycemia inhibits glucose production in man independent of changes in glucoregulatory hormones.

To evaluate the influence of hyperglycemia on hepatic glucose output in the absence of a rise in insulin, glucose was infused for 2 hours into six juvenile-onset diabetics receiving a constant infusion of insulin at a rate of 0.05-0.15 microM kg-1min-1. Prior to the infusion of glucose, insulin administration resulted in stable levels of plasma glucose (76 +/- 8 mg/dl) and glucose output (1.9 +/- 0.1 mg kg-1min-1). The addition of glucose produced a 2-3 fold rise in plasma glucose and a prompt fall in glucose output to 0.2-0.4 mg kg-1min-1, despite the unchanged rate of insulin infusion and the absence of a reduction in plasma glucagon or catecholamines. A similar decline in glucose output was observed when exogenous glucagon (1 ng kg-1-min-1) was added to the glucose infusion. We conclude that in the presence of basal insulin levels hyperglycemia inhibits glucose output independent of a rise in insulin or a fall in anti-insulin hormones.


Increased adrenal sensitivity to angiotensin II in idiopathic hyperaldosteronism.

Plasma aldosterone increases briskly during upright posture in patients with idiopathic hyperaldosteronism, despite only small increases in PRA and presumably small increases in angiotensin II. To examine the postulate that small increments in angiotensin II mediate these brisk increases in aldosterone, we infused graded doses of angiotensin II into normal subjects and patients with idiopathic hyperaldosteronism and compared the changes in levels of plasma aldosterone in the two groups. Supplemental sodium and dexamethasone were given before the infusion to minimize the influence of endogenous angiotensin II and ACTH. In response to the infusion of angiotensin II, increases in the levels of plasma aldosterone of patients with idiopathic hyperaldosteronism were significantly greater than those of normal subjects. In addition, levels of plasma aldosterone increased at a lower rate of infusion of angiotensin II in patients than in normal subjects. It is concluded that patients with idiopathic hyperaldosteronism have increased adrenal sensitivity to angiotensin II. This increased sensitivity may explain the brisk increases in aldosterone that occur during upright posture in these patients.


Adipose tissue lipoprotein lipase in chronic hemodialysis: role in plasma triglyceride metabolism.

The role of adipose tissue lipoprotein lipase (LPL) in the pathogenesis of hypertriglyceridemia in uremic patients receiving maintenance hemodialysis was evaluated. The fasting level of adipose tissue LPL activity was reduced below normal (3.4 +/- 2.5 microU/106 cells; n = 23; mean +/- SD) in hypertriglyceridemic dialysis patients (1.5 +/- 0.8; P less than 0.01; n = 15) and did not differ from normal in normotriglyceridemic dialysis patients (2.5 +/- 2.4; P = NS; n = 13). The enzyme activity increased as a function of relative body weight in normotriglyceridemic hemodialysis patients (r = 0.21; P less than 0.05) but not in the hypertriglyceridemic group (r = 0.21; P = NS). There was an abnormal response of LPL to feeding in the hypertriglyceridemic dialysis patients. The postprandial level of LPL was significantly lower in hypertriglyceridemic dialysis patients (2.2 +/- 1.0; n = 9) than in normotriglyceridemic dialysis patients (3.9 +/- 1.9; P less than 0.05; n = 10) or normal controls (4.8 +/- 1.8; P less than 0.01; n = 12). Whereas the postprandial change in LPL was inversely related to the fasting enzyme activity in normotriglyceridemic dialysis patients (r = 0.74; P less than 0.02; n = 10) and in normal controls (r = 0.58; P less than 0.05; n = 12), no such relationship existed in hypertriglyceridemic dialysis patients (r = 0.17; P = NS; n = 9). Furthermore, fasting plasma triglyceride levels in the entire group of dialysis patients were a function of the postprandial level of LPL activity (rs = 0.574; P less than 0.02; n = 19). Since the level of LPL 1) is below normal in both the fasted and fed state in the hypertriglyceridemic hemodialysis patients, 2) is normal in both the fasted and fed state in the normotriglyceridemic hemodialysis patients, and 3) in the fed state is inversely correlated with the fasting plasma triglyceride concentration in the entire group of hemodialysis patients, it is proposed that adipose tissue LPL plays a role in the etiology of hypertriglyceridemia in hemodialysis patients.


Effect of age on fasting plasma levels of pancreatic hormones in man.

The effect of age and adiposity on fasting plasma levels of pancreatic polypeptide (HPP), glucagon (IRG), insulin (IRI) and glucose was examined in 263 healthy subjects between the ages of 20-69 yr. Mean plasma levels of hPP rose continuously from the third through the seventh decades. Mean plasma levels of IRG rose within the third and fourth decades but failed to rise further thereafter. Mean plasma levels of IRI did not change with age. Mean plasma levels of glucose rose by approximately 2 mg/dl . decade. The correlations of age with hPP, IRG, glucose, and adiposity were 0.47, 0.35, 0.25 (all P less than 0.01) and 0.15 (P less than 0.05), respectively. When adjustments were made for adiposity, the correlations of age with hPP, IRG, and glucose remained. Adiposity correlated with IRI, IRG, and glucose but when age correction was made, only the correlation of adiposity with IRI persisted. We conclude that: 1) age has a significant effect on fasting plasma levels of hPP and IRG; 2) the patterns of the age-related changes in hPP and IRG are not the same, suggesting that there are differences in the mechanism(s) by which age influences plasma levels of these two pancreatic hormones; and 3) age should be considered in the interpretation of fasting plasma levels of hPP and IRG.


Partial purification, characterization, and assay of a slightly acidic insulin-like peptide (ILAs) from human plasma.

An insulin radioreceptor assay (RRA) using human placental microsomal membranes was used to measure insulin-like activity (ILA) extracted from human plasma concentrates (Cohn fraction IV-4) by acid ethanol. The soluble activity (ILAs), chromatographed on Sephadex G-75 in 1 M acetic acid, migrated as a small molecule (fractional elution volume, 0.56) ahead of insulin (fractional elution volume, 0.70), whereas at neutral pH, ILAs migrated as a large molecular weight species. The ILAs peak from acid gel filtration on Sephadex was further purified by chromatography on carboxymethyl cellulose (CMC). The ILAs peak from both Sephadex and CMC diluted parallel to the porcine insulin standard in the insulin RRA and was totally unreactive in an insulin RIA. The CMC-purified material was iodinated and purified by binding to and elution from human placental membranes. The binding of [125I]ILAs to human placental membranes was inhibited only minimally by insulin and proinsulin and not at all by epidermal growth factor, nerve growth factor, glucagon, or lactogenic hormones, including human growth hormone. Multiplication-stimulating activity (MSA) inhibited in a manner parallel to ILAs. A Scatchard plot of the binding data was nonlinear. Sephadex ILAs was subjected to isoelectric focusing. The fractions assayed in both insulin and ILAs RRAs yielded comparable results. Peaks of ILA were observed at pHs 5.3, 6.6, and 8.4. When CMC-ILA was subjected to isoelectric focusing in polyacrylamide, a single peak of activity migrating between pH 6.2-6.8 was seen. [125I]ILAs focused at exactly the same pH. Electrophoresis of CMC-ILAs in acid-urea revealed a sharp peak of activity migrating with one of the five protein bands seen after staining. Again, [125I]ILAs comigrated with unlabeled ILAs. The molecular weight of ILAs, as determined on a calibrated Sephadex G-150 column at neutral pH, was 9,000-10,000 daltons. CMC-ILAs stimulated [14C]glucose incorporation into triglycerides of rat adipose tissue and augmented [3H]thymidine incorporation into human fibroblasts, chicken embryo fibroblasts, and BALB 3T3 cells as well as [35S]sulfate incorporation into macromolecules of rabbit chondrocyte culture medium. In summary, ILAs isolated on the basis of a RRA for insulin is a slightly acidic peptide with some of the biological activities expected of a somatomedin.


Insulin receptors during the menstrual cycle in normal women.

Specific binding of 125I-insulin to circulating monocytes from eight normal menstruating women, four postmenopausal women and four men were studied four times during a 28-day period (one sample at 7-day intervals). Data indicate the presence of a higher specific cell binding fraction in the follicular phase compared to the luteal phase due to changes in insulin receptor concentration. No changes were observed in men or postmenopausal women during the same period of time suggesting that sex hormones should be included among the factors influencing insulin receptors.


Effects of galactose, glucose and fructose on carbohydrate-lipid metabolism.

Doses of glucose, galactose and fructose were administered orally and intravenously, while monitoring the levels of blood glucose, serum insulin, free fatty acids, cholesterol and triglycerides. Blood glucose increases when glucose is administered orally or intravenously, but does not vary significantly with any other sugar. The increase in insulinemia only with oral and intravenous glucose and with oral galactose is in agreement with the participation of intestinal factors in the determination of insulin secretion. The results obtained for free fatty acids with the various sugars may be due to the metabolic properties of each sugar. We found no significant variations in the levels of cholesterol and triglycerides, perhaps because of the short duration of the experiment.


Burns: early shaving and immediate skin-grafting of dorsum of the hand.

Fourteen men with deep dermal burns of the dorsum of the hand treated by early excision and primary skin grafting are presented. Satisfactory functional and cosmetic results within two to three weeks were obtained in ten patients and in five to six weeks in the remaining four. Three patients who refused surgery developed contractures. The procedure is recommended as a routine.


Neutrophil antigens: immunology and clinical implications.

Neutrophil antigens may be classified into two major categories: antigens shared with other cells and antigens specific for neutrophils. The first category includes the ABH, I, i, 5a,b and HLA determinants. Additional antigens with special characteristics in this category are the blood-group U, Kx, JkaJkb, and Ge determinants which apparently neutrophils share only with erythrocytes. Neutrophil-specific antigens include the NA1, NA2, NB1 and 9a. These specificities are detected by the agglutination test and have been shown to be present on mature neutrophils. Independent allospecificities, detectable by the granulocytotoxicity test, may also exist. In addition, neutrophil antigens, which are species-specific, have been identified by the use of xenogeneic antibodies. The EDTA-dependent agglutination test remains a most reliable assay for the study of neutrophil-specific antigens. The lack of reproducibility known in the leukoagglutination reaction does not pertain to the modification used in the assay of neutrophil-specific antibodies. It does apply, however, to those tests that were performed in the absence of EDTA, and in connection to the study of HLA-related antigens. For every pathophysiological state involving the erythrocyte antigens a neutrophil analogue is observed, the difference being in symptomatology which is related to the structural and functional characteristics of the cells: febrile and pulmonary transfusion reactions result from incompatibility neutrophils. It is found that similarity in the HLA antigens and nonreactivity in the MLC test do not preclude immunization against neutrophil-specific antigens. Therefore, it is probable that febrile and pulmonary reactions will occur in the recipients of multiple granulocyte transfusions, even though donors and recipients may be considered "histocompatible" by the HLA assays. It has been shown that fetal-maternal incompatibility can cause neonatal neutropenia, and several forms of autoimmune neutropenia are described: in "idiopathic" neutropenia of infancy, autoantibodies have been found to have specificity against NA1 and NA2 and in one adult, autoimmune neutropenia due to anti-NA1 antibody has been observed. Neutropenia also occurs due to idiopathic, cold-reacting antileukocyte antibodies, and with cold agglutinins associated with lymphoma, infectious mononucleosis, and Mycoplasma pneumonia. Although the role of neutrophil antigens in bone marrow transplantation has not as yet been determined, these antigens are undoubtedly immunogenic and potentially play an important role in neutrophil compatibility. It is obvious that neutrophils cannot survive in the presence of antineutrophil antibodies.(ABSTRACT TRUNCATED AT 400 WORDS)


The regulation of the release of granulocytes from normal marrow.

Figure 16 synthesizes the various aspects of marrow egress. The central anatomical relationship of the hematopoietic compartment to vascular sinus is shown above. The hatched blocks represent the sinus wall, capable of developing narrow migration channels. In the marrow, immature granulocytes alter their biophysical characteristics by developing motility, nuclear and cytoplasmic deformability and surfaces which facilitate egress. Humoral factors contribute to proliferation and maturation. Also, other humoral agents, releasing factors, may act on mature cells, for example, as cytoattractants, and on the sinus wall to reduce its adventitial cover and thereby to enhance egress. In the sinus, flow or discharge of sinus contents may be regulated by humoral agents or neural messages which may affect terminal sphincters or other structures. Although the marrow in situ is a difficult organ to study, future innovations are to be expected and our understanding of the delicate balance between hematopoietic cells, stroma and vasculature will be enhanced. Corrections of inferential errors due in part to the semi-quantitative and qualitative nature of much of our current data should be expected.


Mosaicism in amniotic fluid cell cultures: classification and significance.

The last decade has witnessed increasing application of human cytogenetic technology to prenatal chromosome analysis. However, unlike the rather uniform peripheral blood T-lymphocyte system which has provided most of our experience in human cytogenetics, long-term amniotic-fluid cell cultures display extreme cellular heterogeneity and disproportionate growth of certain cell types as a consequence of clonal amplification. When they enter cell culture, many of these cells are approaching the terminal stages of their respective, life spans and may have accumulated chromosomal aberrations. Concern about the possibility of true fetal mosaicism seems warranted chiefly in situations were multiple colonies display potentially viable aberrations. Clonal analysis, preferably of multiple clonal types, and attention to details of clonal morphology are likely to minimize diagnostic errors and undue apprehension resulting from mosaicism in amniotic-fluid cell cultures.


National Health Insurance: a political issue at the crossroads.

It is safe to assume that a comprehensive NHI plan will not be enacted prior to the next presidential election. In sum, immediate action on the NHI issue is unlikely. However, enactment of a mini-NHI plan, expanding the Medicare and Medicaid programs, providing catastrophic coverage and/or providing coverage for mothers and children, is possible--although not probable--prior to the next presidential election.


Mechanisms of impaired acute insulin release in adult onset diabetes: studies with isoproterenol and secretin.

Previous work has suggested that impaired islet glucose recognition occurs in patients with adult onset diabetes, as acute insulin release is absent after iv glucose but present after beta adrenergic stimulation with isoproterenol (Iso). However, insulin responses to Iso were variably reduced as compared to normal in the diabetics. In order to evaluate the importance of the Iso dose, dose-response studies were performed in 9 diabetics (fasting plasma glucose greater than 150 mg/dl) and 10 age-matched controls. In both control subjects and diabetics, 0.5 microgram Iso produced no insulin response; 2 micrograms Iso produced an intermediate response; and 8 and 12 micrograms Iso produced a higher response. The insulin responses to the larger doses of Iso were lower in diabetics than control subjects (8 micrograms, 20 +/- 5 vs. 39 +/- 6 (P less than 0.025); 12 micrograms, 21 +/- 6 vs. 37 +/- 4 (P less than 0.05); means +/- SEM, microU/ml). Of 16 diabetics who received 12 micrograms Iso, 5 had insulin responses greater than 2 SD below the control mean, while others had responses that spanned the entire range of normal. Seven diabetics also were given iv secretin (150 U). Their insulin responses to secretin correlated with the responses to Iso (r = 0.83, P less than 0.02). Thus, patients with subnormal responses to Iso also had low secretion responses. The abnormalities of acute insulin secretion in diabetics can be explained by a lesion variably affecting islet membrane receptors; some patients may have glucose receptor damage, but intact responses to other stimuli, and others may have more widespread damage affecting beta-adrenergic and secretin responses as well. Alternatively, there may be heterogeneity in adult onset diabetes, as patients with low responses to all stimuli could have a qualitatively different lesion affecting insulin secretory capacity rather than membrane receptors.


[General anesthesia in infants].

It is generally accepted, that the progress in surgical treatment of congenital malformation is closely related to the progress in highly-specialised methods of newborn anaesthesia. The safe methods of anaesthesia have to be adapted to anatomical and physiological peculiarities of the earliest days of life as well as to different reaction to drugs and anaesthetics. The preoperative preparation in newborns used to be often very short, because most of the problems of the neonatal period are emergency surgical interventions and there is no time for treatment even of the serious disturbances of basic physiological functions. The purpose of this study was to estimate methods of general anaesthesia in newborns, which have been introduced in the Anaesthesiology Department of the National Research Institute of Mother and Child. The main element of these methods was general anaesthesia with muscle relaxants and controlled ventilation as a routine. The estimation was based on general analysis of 10 years practice when these methods of anaesthesia were used. During this time 515 anaesthesias to 408 both-sex newborns were given (Tab. I, Fig. 1). 85% of anaesthetized newborns were operated because of congenital malformations (Tab. I); 46% of operations were performed during their first week of life, 21% were operated in first 24 hours of life, mostly as an emergency (Fig. 3). 10% of operations were performed is prematures (body weight below 2500 g) (Fig. 4). The "routine" anaesthesia was given in 82% cases. Awake intubation in unpremedicated newborns was performed. Anaesthesia was maintained with nitrous oxide-oxygen mixture (1:1 or 2:1). D-tubocurarine in 95% of cases was used. The initial dose 0,5 with matures and 0,25 mg with premature babies was used. If necessary supplementary doses were given. During anaesthesia, intermittent positive pressure ventilation (IPPV) with frequency at least 60/min. was used. During this ventilation, hyperventilation and positive end-expiratory pressure (PEEP) were obtained. Precordial stetoscope and thermometer probe was used as a routine. In some special cases eCG, end-expired CO2 (capnography), pletysmography were also recorded; blood gas analyses were checked. All intra- and postoperative complications as well as postoperative mortality have been analysed in details. During 3,3% of operations some complications had been observed. The total incidence of early psotoeprative complications was 20%. In this group the most frequent were respiratory complications (16,1%). Serious disturbances in pulmonary gas exchange during operation and early postoperative period were not found.(ABSTRACT TRUNCATED AT 400 WORDS)


[Chemotherapy of bladder and prostatic neoplasms in children].

The basic informations concerning pathology and clinical stagging of malignant tumours localized in urinary bladder and prostatic gland are introduction to proper presentation of own material collected and treated in the years 1962-1977. The material consists of 23 cases of rhabdomyosarcoma localized in minor basin. Formerly in the years 1962-1975, the basic form of treatment was surgery with/or radiotherapy. Chemotherapy that was given accidently in special indications as single dose or for a short time. Since 1975, we have started with systematic multidrug chemotherapy (VAC) used as an important part of complex therapy. Finally the authors discuss the place of chemotherapy in complementary treatment on the base of the literature and their own experience.


Factors influencing the quantity and quality of semen harvested from bulls, rams, boars and stallions.

Several inherited conditions associated with testicular defects, abnormal spermatogenesis and morphologically abnormal sperm have been found. These usually are controlled by single gene pairs. A notable exception is testicular size, with heritability in young bulls ranging from .42 to .88. Testicular size directly affects sperm output potential. The major contributor to variation in semen quality is the environment. Environmental effects may be temporary or permanent. Permanent effects occurring during prenatal and prepubertal periods and temporary or permanent factors acting after spermatogenesis is initiated can alter semen quality. Semen quality improves during the first few months after puberty and declines in old age. Malnutrition and the ingestion of toxic materials can have a major effect on testicular development and spermatogenesis, but the reproductive system has considerable regenerative capacity unless the dietary deficiencies are severe and prolonged. Elevated testicular temperatures resulting from incomplete descent of the testes (cryptorchidism), high environmental temperatures or inflammation are detrimental to spermatogenesis in all scrotal mammals. Cold temperatures appear to be innocuous unless actual freezing of tissue occurs. During periods of decreasing daylight semen quality declines in stallions and improves in seasonally breeding sheep. The time required to form and transport sperm in bulls, rams, boars and stallions is about 64, 56, 47 and 59 days. Therefore, a considerable potential lag may exist between a testicular event responsible for a change in semen quality and the time that this change is evident in ejaculated semen. Conditions imposed at the time of semen collection such as frequency of ejaculation, degree of sexual preparation and type of semen collection may influence the quality of semen harvested. Finally, certain semen characteristics are more variable than others and investigators should utilize suitable existing data to design the most effective least-cost experiments.


Selection among males for the genetic improvement of female fertility.

The quantitative physiological study of genetic variation in reproductive performance has shown that differences in the litter size and incidence of lambing in sheep are associated with variation in the release of LH. This variation is detectable in young animals of both sexes, and the injection of LHRH may aid its recognition. In the male it is associated with variation in mating behavior and testis growth. The ovarian activity of both sheep and mice has been shown to change following selection for testis growth. The use of measures of testis growth as criteria for selection to improve the reproductive performance of females is discussed.


Gametogenic processes and their relationship to normal and abnormal conceptus development.

The relationship between gametogenesis and conceptus development has been described in relation to recent experiments concerning selected aspects of the cellular and molecular basis of differentiation of female and male gametes. Evidence suggests that a variety of hormonal and nonhormonal mechanisms or combinations thereof have evolved and control distinct stages of oocyte maturation process. Analyses of the meiotic maturation process with particular emphasis on amphibians, indicate that distinct cytoplasmic factors are produced or activated in response to a hormonal stimulus which initiates germinal vesicle breakdown and reinitiates the meiotic maturation process. Evidence suggests that certain cytoplasmic factors originate in the nucleus and (or) cytoplasm and that considerable cytoplasmic maturation proceeds in the absence of the nucleus, or as a result of nuclear-cytoplasmic interactions. Cytoplasmic factor(s) or activities affect many aspects of oocyte function and structure, including the cell membrane, chromosomes, nucleus and meiotic spindle. Ions also play a crucial role in gamete differentiation either alone or in combination with hormones or these cytoplasmic factors. The relevance of these findings to oocyte maturation and fertilization, activation and embryonic development is discussed. In many cases, similar types of biological activities exist in widely separated species and (or) are effective in widely different species. The importance of synchronization of normal gametes to the development of the embryos is discussed. It is proposed, and the evidence suggests, that hormones, ions and cytoplasmic factors play a fundamental and variable role in the differentiation and function of "fully" mature oocytes of many species including mammals. Significantly, the data suggest that a "fully" mature oocyte occurs as a result of the expression of an underlying developmental program. Variations in these substances or the processes involved in their formation or actions appear to be relevant to understanding a wide variety of developmental abnormalities as well as to assessing the normality or abnormality of in vivo or in vitro maturation and differentiation.


Actions of hormones on the uterus and effect on conceptus development.

The process of reproduction in the mammalian female involves interactions between gonadal and placental steroids and, possibly, protein hormones such as placental lactogen, and the uterine endometrium. These interactions are discussed relative to available data from pigs and with the understanding that the concepts presented may not be directly applicable to other species. Swine embryos enter the uterus for continued development beyond the early blastocyst stage and, within the uterus, the spherical blastocyst, of about 2 mm diameter on day 12, undergoes rapid elongation to an organism of about 1 m in length by day 15 of pregnancy. During this period of elongation, the blastocyst appears to provide the chemical signal, presumably unconjugated estrogens, that allows for corpora lutea (CL) maintenance (luteostatic effect) and, therefore, continued production of progesterone by the CL which is essential for pregnancy. Estrogens from the blastocyst may exert their luteostatic effect on the uterus by causing prostaglandin F2 alpha, the presumed porcine uterine luteolysin, to be secreted in an exocrine (toward the uterine lumen) rather than an endocrine (toward the endometrial stroma and vasculature) direction during pregnancy. Consequently, prostaglandin F2 alpha is sequestered in the uterine lumen and does not become available, via the utero-ovarian vasculature, to exert its luteolytic effect. The potential steroid precursor(s) available to the trophoblast for estrogen synthesis appear to be produced through endometrial conversion of progesterone to unconjugated androgens, e.g., androstenedione and testosterone, and conjugated estrogens, e.g., estrone-sulfate and estradiol-sulfate. The estrogens may, in concert with progesterone and possibly other hormones, placental lactogen for example, act locally, i.e., at the site of placentation to: result in a luteostatic effect; increase uterine blood flow; enhance water, electrolyte and substrate (carbohydrate and amino acid) transport to the site of placentation and across the placenta; affect synthesis and secretion of macro-molecules (histotroph) by the uterine glands that serve as enzymes and (or) carrier molecules and control or modulate physiological and (or) biochemical events essential to placental and fetal development that are not understood.


Mechanism of action of estrogen agonists and antagonists.

The relationship between estrogen agonists and antagonists and the nuclear binding of the estrogen receptor was investigated in the immature rat uterus. Estriol (and other short acting estrogens) are antagonistic when administered as a single injection due to the inability of the receptor-estriol complex to be retained by uterine nuclei for a critical period necessary to stimulate true uterine growth. This failure of the receptor-estriol complex to be retained appears to be due to rapid clearance of estriol from uterine tissue following a single injection of the hormone. When estriol is present in a chronic fashion, it acts as an agonist because receptor estriol complex is retained at nuclear sites for long periods of time. Nonsteroid estrogen antagonists, such as Nafoxidine, also cause long term nuclear retention of the estrogen receptor and act as estrogen agonists after a single injection. However, after multiple injections these drugs are antagonistic and this effect is correlated with decrease availability of cytoplasmic estrogen receptors. In addition, these compounds appear to specifically stimulate the growth of uterine epithelial cells while minimally affecting other cell types which results in hyperestrogenization of the epithelium.


Genetic variation in prenatal survival and litter size.

The nature of genetic variation in litter size and its components was examined using results of long term selection and of crossing selected lines of mice. Selection for small and for large litters produced strains with mean litter size of approximately 6 and 15, respectively. The large litter size line showed a period of renewed response after 45 generations of selection, following a period of 15 generations with no response. The relative contribution of genotype of the dam and of the embryo to prenatal survival was examined using results of selection, crossing and embryo transfer. Genotype of female carrying the litter was responsible for most of the observed differences. Genes for poor survival expressed by the embryo were present in some strains, but accounted for less variation than did maternal genotype. Information from the selected lines and from crosses among them provided evidence for dominance of genes affecting fertility and prenatal survival and additivity of genes affecting ovulation rate and body weight. The information was used to suggest a model for more effective exploitation of heterosis using breeds or strains of animals of known performance for different components. The extent of genetic variation for prenatal survival suggests that natural selection in this species may not always be for maximum survival. It is suggested that the ability to reduce litter size after conception, in response to nutritional or other stress, may at times be of selective advantage, contributing to maintenance of genetic variation in survival rate.


Psychological treatment of cardiovascular disease.

Recent clinical research on the psychological treatment of cardiovascular diseases is reviewed in four categories: hypertension, cardiac arrhythmias, coronary artery disease, and peripheral circulatory disease. In the treatment of hypertension biofeedback of blood pressure, electromyography and galvanic skin responses both seem useful, as does systematic relaxation training of either an active or passive-meditative nature. Biofeedback of heart rate has shown some utility in treating premature ventricular contractions and sinus tachycardia. Supportive and educational group therapy for patients recovering from myocardial infarctions has shown some utility. In the treatment of Raynaud's disease, biofeedback of skin temperature is helpful. In no area has a large scale clinical trial been conducted, and only three controlled group outcome studies have been reported (two in hypertension, one in coronary artery disease). Overall, the evidence is impressive enough to warrant more systematic controlled investigation in all four areas.


Relaxation therapy in the treatment of hypertension. A review.

The literature on the use of relaxation or relaxation-like procedures (relaxation therapy) in the treatment of hypertension was critically reviewed. Relaxation therapy resulted in greater reduction of blood pressure than placebo or other control procedures. A positive relationship was found between the average blood pressure decrease and the average pretreatment pressure. Relaxation-like therapies shared the features of muscular relaxation, regular practice, mental focusing, and task awareness. Research on the relative contributions of these components indicated that task awareness adds to the treatment effect in the laboratory setting, and that regular practice is necessary for optimal results in the clinical setting. The role of muscular relaxation and mental focusing is unclear. We concluded that relaxation therapy may become a useful adjunct to medication in the clinical management of hypertension, especially for individuals whose blood pressures remain high despite pharmacological treatment.


Reactions of transcendental meditators and nonmeditators to stress films. A cognitive study.

To experimentally test the claimed stress-reducing effects of Transcendental Meditation (TM), two stress films were shown to a group of 60 meditators and nonmeditators. Stress response was observed through the use of cognitive and affective measures employing content analysis techniques and self-ratings. The meditators did not show less stress response than the nonmeditators. On several self-rating scales, a group of subjects who had signed up to be initiated into TM rated themselves significantly more stressed and emotionally distressed than either a control group or meditators. There was a trend for meditators who meditated during the experiment to show less stress response to the films than meditators who were told not to meditate; however, this difference was significant on only one measure, a subjective stress scale.


Suppression of the DnaA phenotype by mutations in the rpoB cistron of ribonucleic acid polymerase in Salmonella typhimurium and Escherichia coli.

A class of mutations that confer resistance to rifampin in Salmonella typhimurium and Escherichia coli also suppresses the thermosensitivity of chromosome initiation in dnaA mutants. Ribonucleic acid polymerase is resistant to rifampin in vitro in these suppressive mutants, and the suppressors of dnaA cannot be separated from the rpoB mutations by transduction. It is concluded, therefore, that certain rpoB mutations may suppress the DnaA phenotype.


Influence on motility of Escherichia coli and Salmonella typhimurium by a naturally occurring conjugative plasmid.

In a collection of 45 R-plasmids, one was found to be associated with loss of motility of its Escherichia coli K-12 and Salmonella typhimurium host bacteria when tested in conventional motility agar. Genetic experiments, as well as analyses of deoxyribonucleic acid, showed that inhibition of motility was caused by a conjugative plasmid that was separate from the R-plasmid. This second plasmid, named pUM5, was fi- and mediated the same type of sex pilus (F-like) as the accompanying R-plasmid but lacked resistance determinants. Preliminary studies indicated that bacterial cells carrying the motility inhibition plasmid pUM5 were still equipped with flagella. The mechanism by which flagellar action is disturbed by the plasmid is presently not known.


In vitro transcription of the Escherichia coli K-12 argA, argE, and argCBH operons.

Deoxyribonucleic acid isolated from argA and argECBH transducing phages was utilized to study the in vitro synthesis of argA, argE, and argCBH messenger ribonucleic acid. The specific regulation of these operons by the arginine holorepressor was demonstrated, providing evidence that the majority, if not all, of the control of these operons is exercised at the transcriptional level. Data are presented which indicate that the arginine holorepressor functions by binding to the operator region and concomitantly prevents the binding of ribonucleic acid polymerase to the corresponding promoter region.


Genetic analysis of Escherichia coli O111:B4, a strain of medical and biochemical interest.

Procedures have been worked out which allow, for the first time, the genetic analysis of Escherichia coli O111:K58:H2 (O111:B4). The approximate map position of mutant loci was determined by mating with 15 Hfr strains of E. coli K-12. In addition, P1 transduction procedures were used for establishing relative gene order and linkage for any region of the E. coli O111:B4 chromosome. To obtain these, it was necessary to select for a rare P1 lysogen since E. coli O111:B4 is resistant to phage P1. Finally, genetic homology between E. coli strains K-12 and O111:B4 is suggested since they can form stable haploid hybrids, and several loci have similar map positions in the two strains.